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Небесная энциклопедия

Космические корабли и станции, автоматические КА и методы их проектирования, бортовые комплексы управления, системы и средства жизнеобеспечения, особенности технологии производства ракетно-космических систем

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Мониторинг СМИ

Мониторинг СМИ и социальных сетей. Сканирование интернета, новостных сайтов, специализированных контентных площадок на базе мессенджеров. Гибкие настройки фильтров и первоначальных источников.

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Поддерживает ввод нескольких поисковых фраз (по одной на строку). При поиске обеспечивает поддержку морфологии русского и английского языка
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Применить Всего найдено 353. Отображено 100.
19-01-2012 дата публикации

PARTICLE-ASSISTED NUCLEIC ACID SEQUENCING

Номер: US20120015822A1
Автор: Abate Adam R., Weitz David A.
Принадлежит: President and Fellows of Harvard College

This invention generally relates to particle-assisted nucleic acid sequencing. In some embodiments, sequencing may be performed in a microfluidic device, which can offer desirable properties, for example, minimal use of reagents, facile scale-up, and/or high throughput. In one embodiment, a target nucleic acid may be exposed to particles having nucleic acid probes. By determining the binding of the particles to the target nucleic acid, the sequence of the target nucleic acid (or at least a portion of the target nucleic acid) can be determined. The target nucleic acid may be encapsulated within a fluidic droplet with the particles having nucleic acid probes, in certain instances. In some cases, the sequence of the target nucleic acid may be determined, based on binding of the particles, using sequencing by hybridization (SBH) algorithms or other known techniques. 1. A method of sequencing a target nucleic acid , the method comprising:exposing a target nucleic acid to a plurality of particles, at least some of which have nucleic acid probes fastened thereto; andat least partially determining the sequence of the target nucleic acid based on binding of the particles to the target nucleic acid.2. The method of claim 1 , wherein at least one of the particles contains at least one identification entity.3. The method of claim 2 , further comprising at least partially determining the sequence of the target nucleic acid by determining the at least one identification entity.4. The method of claim 2 , further comprising determining the at least one identification entity.57-. (canceled)8. The method of claim 1 , wherein at least some of the nucleic acid probes contain at least six residues.914-. (canceled)15. The method of claim 1 , wherein the target nucleic acid comprises a signaling entity.16. The method of claim 1 , further comprising determining association of the target nucleic acid and at least some of the particles.1719-. (canceled)20. The method of claim 1 , further ...

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Номер записи: 1
03-05-2012 дата публикации

Systems and methods of templating using particles such as colloidal particles

Номер: US20120107601A1
Автор: André R. Studart, David A. Weitz, James N. Wilking, Jens B. Rieger, Roland S. Koltzenburg, Roland S. Weitz
Принадлежит: BASF SE, Harvard College

The present invention generally relates to systems and methods for using particle templating, e.g., to produce composites, discrete particles, or the like. In some embodiments, the present invention generally relates to the production of particles using the interstitial spaces between templating elements in a template structure. For example, a plurality of templating elements, which can include colloidal particles, may be arranged to form a template structure. The interstices of the templating elements can provide regions in which a fluid may be introduced. The fluid may be hardened (e.g., solidified) in some cases, e.g., to form a composite comprising the templating elements and the interstitial segments. In certain embodiments, the template structure may then be broken down to release the hardened fluid, e.g., as a plurality of discrete particles.

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Номер записи: 2
07-06-2012 дата публикации

PARTICLES FOR DRUG DELIVERY AND OTHER APPLICATIONS

Номер: US20120141589A1
Автор: Fan Ben, Koltzenburg Roland S., Rieger Jens B., Weitz David A.
Принадлежит:

The present invention generally relates to particles for drug delivery and other applications. In one aspect, the present invention relates to a technique for reacting precursor compounds in the presence of a pharmaceutically-active agent to form product (e.g., in the form of particles) in which the agent is substantially contained within the product, and the product is soluble within typical gastric fluid of a mammal. In another aspect, the present invention is generally directed to particles comprising an inorganic pharmaceutically acceptable carrier, such as CaCO, and an agent. In some cases, at least some of the agent contained within the particles is fluidically inaccessible from externally of the particle. For instance, the agent may be present in isolated domains within the particle. In another aspect, the present invention is generally directed to methods of creating particles. For instance, according to one set of embodiments, two fluids containing reactants are mixed where, upon reaction of the reactants, an insoluble product is formed, which precipitates to form particles. In one example, a first fluid containing dissolved carbonate ions and a second fluid containing dissolved calcium ions and a pharmaceutically-active agent are mixed together; upon mixing of the first and second fluids, the calcium ions and the carbonate ions form calcium carbonate, which precipitates to form a co-precipitate with the pharmaceutically-active agent. Yet other aspects of the present invention are directed to particles formed from such reactions, methods of using such reactions, methods of promoting such reactions, kits involving particles, or the like. 14-. (canceled)5. The composition of claim 20 , wherein the particle has a largest dimension of no more than about 100 micrometers.6. The composition of claim 20 , wherein the agent has a solubility in water of less than about 10 g/l at 20° C. and 1 bar.7. The composition of claim 20 , wherein at least one of the one or more ...

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Номер записи: 3
05-07-2012 дата публикации

SPRAY DRYING TECHNIQUES

Номер: US20120167410A1
Автор: Abate Adam R., HOLTZE Christian, Thiele Julian W.P., Weitz David A., Windbergs Maike
Принадлежит:

The present invention generally relates to microfluidics, and to spray drying and other drying techniques. In some aspects, an article containing one or more channels or microfluidic channels is used to mix one or more fluids prior to spray drying. The mixing may occur immediately before the fluids are expelled through a nozzle or other opening into a drying region of the spray dryer. In one set of embodiments, for example, a first fluid is exposed to a second fluid, then the fluids are exposed to air or other gases before being expelled through a nozzle. In certain instances, the first fluid may contain a dissolved species that may precipitate upon exposure to the second fluid; such precipitation may occur immediately before expulsion through a nozzle or other opening, thereby resulting in controlled precipitation as part of the spray drying process. 1. A spray dryer for use in drying fluids , comprising: a first microfluidic channel having an opening as a nozzle; and', 'a second microfluidic channel intersecting the first microfluidic channel at an intersection upstream of the nozzle; and, 'an article comprisinga drying region that receives output from the nozzle.2. The spray dryer of claim 1 , wherein the article further comprises a first fluid source in fluid communication with the first microfluidic channel.3. (canceled)4. The spray dryer of claim 1 , wherein the article further comprises a second fluid source in fluid communication with the second microfluidic channel.5. (canceled)6. The spray dryer of claim 4 , wherein the article further comprises a third microfluidic channel in fluid communication with the second fluid source claim 4 , the third microfluidic channel intersecting the first microfluidic channel at the intersection upstream of the nozzle.7. The spray dryer of claim 1 , wherein the article further comprises a fourth microfluidic channel intersecting the first microfluidic channel at a second intersection upstream of the intersection of the ...

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Номер записи: 4
09-08-2012 дата публикации

MULTIPLE EMULSIONS CREATED USING JUNCTIONS

Номер: US20120199226A1
Автор: HOLTZE Christian, Romanowsky Mark, Weitz David A.
Принадлежит:

The present invention generally relates to emulsions, and more particularly, to multiple emulsions. In one aspect, multiple emulsions are formed using a plurality of channels, such as microfluidic channels, that meet at a common intersection. The multiple emulsions may be created at a single common intersection in some embodiments, unlike other prior art systems where multiple channel intersections are required to create multiple emulsions. For instance, in one set of embodiments, three, four, or more microfluidic channels may intersect at a common intersection, with two or three serving as inlets and one serving as the outlet. In some embodiments, a first fluidic channel may be relatively hydrophobic, while a second fluidic channel is relatively hydrophilic. The third channel, if present, may be relatively hydrophilic or hydrophobic, depending on the application. The outlet channel may be hydrophobic, hydrophilic, or may comprise at least one portion that is relatively hydrophilic and at least one portion that is relatively hydrophilic. By controlling the flow of fluids through the hydrophilic and hydrophobic portions of the channels, multiple emulsions may be created proximate the common intersection, due to interactions between the fluids entering the common intersection. In other embodiments, different patterns of hydrophilic or hydrophobic channels may be used. Other aspects of the invention are generally directed to methods of making and using such systems, kits involving such systems, emulsions created using such systems, or the like. 1. A device , comprising:at least first, second, third, and fourth fluidic channels intersecting at a common intersection, wherein the first fluidic channel is relatively hydrophobic, the second fluidic channel is relatively hydrophilic, and the fourth fluidic channel comprises at least one portion that is relatively hydrophilic and at least one portion that is relatively hydrophilic.2. A device , comprising:at least first, ...

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Номер записи: 5
23-08-2012 дата публикации

MULTIPLE EMULSIONS CREATED USING JETTING AND OTHER TECHNIQUES

Номер: US20120211084A1
Автор: Abate Adam R., Thiele Julian W.P., Weitz David A.
Принадлежит: President and Fellows of Harvard College

The present invention generally relates to emulsions, and more particularly, to multiple emulsions. In one aspect, multiple emulsions are formed by urging a fluid into a channel, e.g., by causing the fluid to enter the channel as a “jet.” Side channels can be used to encapsulate the fluid with a surrounding fluid. In some cases, multiple fluids may flow through a channel collinearly before multiple emulsion droplets are formed. The fluidic channels may also, in certain embodiments, include varying degrees of hydrophilicity or hydrophobicity. As examples, the fluidic channel may be relatively hydrophilic upstream of an intersection (or other region within the channel) and relatively hydrophobic downstream of the intersection, or vice versa. In some cases, the average cross-sectional dimension may change, e.g., at an intersection. For instance, the average cross-sectional dimension may increase at the intersection. Surprisingly, a relatively small increase in dimension, in combination with a change in hydrophilicity of the fluidic channel, may delay droplet formation of a stream of collinearly-flowing multiple fluids under certain flow conditions; accordingly, the point at which multiple emulsion droplets are formed can be readily controlled within the fluidic channel. In some cases, the multiple droplet may be formed from the collinear flow of fluids at (or near) a single location within the fluidic channel. In addition, unexpectedly, systems such as those described herein may be used to encapsulate fluids in single or multiple emulsions that are difficult or impossible to encapsulate using other techniques, such as fluids with low surface tension, viscous fluids, or viscoelastic fluids. Other aspects of the invention are generally directed to methods of making and using such systems, kits involving such systems, emulsions created using such systems, or the like. 1. An apparatus , comprising:a main microfluidic channel;at least one first side microfluidic channel ...

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Номер записи: 6
21-02-2013 дата публикации

CONTROL OF EMULSIONS, INCLUDING MULTIPLE EMULSIONS

Номер: US20130046030A1
Автор: Abate Adam R., HOLTZE Christian, Rotem Assaf, Weitz David A.
Принадлежит:

The present invention generally relates to emulsions, and more particularly, to double and other multiple emulsions. Certain aspects of the present invention are generally directed to the creation of double emulsions and other multiple emulsions at a common junction of microfluidic channels. In some cases, the microfluidic channels at the common junction may have substantially the same hydrophobicity. In one set of embodiments, a device may include a common junction of six or more channels, where a first fluid flows through one channel, a second fluid flows through two channels, and a third or carrying fluid flows through two more channels, such that a double emulsion of a first droplet of the first fluid, contained in a second droplet of the second fluid, contained by the carrying fluid, flows away from the common junction through a sixth channel. 1. A microfluidic device , comprising:a first junction of microfluidic channels comprising at least first, second, and third microfluidic channels in fluidic communication, the first junction in fluid communication at an interface with a second junction of microfluidic channels comprising at least fourth, fifth, and sixth microfluidic channels in fluidic communication, each of the first, second, and third microfluidic channels having a respective cross-sectional area at the first junction and each of the fourth, fifth, and sixth microfluidic channels having a respective cross-sectional area at the second junction, wherein the interface has a cross-sectional area smaller than the smallest cross-sectional areas of the fourth, fifth, and sixth microfluidic channels.2. The microfluidic device of claim 1 , wherein the sixth microfluidic channel defines a central axis claim 1 , and the interface has a center point that is substantially on the central axis of the sixth microfluidic channel.3. The microfluidic device of claim 1 , wherein the first microfluidic channel defines a first central axis and sixth microfluidic channel ...

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Номер записи: 7
14-03-2013 дата публикации

SYSTEMS AND METHODS FOR SHELL ENCAPSULATION

Номер: US20130064862A1
Автор: HOLTZE Christian, Shum Ho Cheung, Sun Bingjie, Weitz David A., Zhao Yuanjin
Принадлежит:

Certain aspects of the invention are generally directed to particles comprising a shell and an interior at least partially contained by the shell. In some embodiments, the particles may be treated to enhance the containment of the interior, for example to reduce transport of an agent into or out of the interior. Such particles may exhibit increased ability to encapsulate agents and/or increased storage life (e.g., due to reduced leakage). For instance, in certain embodiments, any defects, such as cracks, pores, etc. within the shell may be sealed or otherwise treated to reduce transport therethrough. In some embodiments, for instance, a first reactant in the interior of a particle may come into contact with a second reactant outside of the particle to form a solid, or other suitable product. The shell may also be treated to cause release of an agent contained within the interior, in certain aspects. 1. An article , comprising:a fluid containing a microparticle comprising a shell formed from a shell material and an interior at least partially contained by the shell, the interior containing a first reactant and the fluid containing a second reactant, wherein one or both of the first reactant and the second reactant is able to move towards the other, and wherein the first reactant and the second reactant are able to react to form a product.2. The article of claim 1 , wherein one or both of the first reactant and the second reactant is able to move into or through the shell to form the product.3. The article of claim 1 , wherein the product is not able to substantially move out of the shell.4. The article of claim 1 , wherein the product is substantially insoluble in the fluid and the interior.5. The article of claim 1 , wherein the product is a solid.6. The article of claim 1 , wherein the product is a salt.7. The article of claim 1 , wherein the product comprises CaCO.8. The article of claim 1 , wherein the shell comprises a polymer.9. The article of claim 1 , wherein ...

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Номер записи: 8
08-08-2013 дата публикации

FOAMS OR PARTICLES FOR APPLICATIONS SUCH AS DRUG DELIVERY

Номер: US20130202657A1
Автор: Guerra Rodrigo E., Kaz David, Koltzenburg Roland S., Ladavac Kosta, Manoharan Vinothan, Rieger Jens B., Weitz David A.
Принадлежит:

The present invention generally relates to foams and, in particular, to foams for applications such as drug delivery, and particles that are made from such foams. One aspect relates to foams or particles containing pharmaceutically active agents. The foam may comprise a pharmaceutically acceptable polymeric carrier. In some cases, the foam or particle has an unexpectedly high specific surface area. A high specific surface area may, in some cases, facilitate delivery or release of the pharmaceutically active agent when the foam or particles made from the foam (e.g., by milling) are administered to a subject. The foam may also exhibit a relatively high loading of the pharmaceutically active agent. In some cases, the foam may be a microcellular foam. In one set of embodiments, the foam is created using a supercritical fluid, such as supercritical C02. For example, a precursor to the foam, containing a pharmaceutically active agent, may be mixed with a foaming agent, then the pressure decreased to cause the foaming agent to expand, thereby causing a foam to form. The foam may then be subsequently ground or milled, or otherwise processed to form particles. 1. A pharmaceutically active article , comprising:{'sup': 2', '3, 'a foam comprising a pharmaceutically acceptable polymeric carrier and a pharmaceutically active agent, the foam having an average cell size of less than about 5 micrometers, wherein the foam (a) has a specific surface area of at least about 0.4 m/g, and/or (b) has a foam density of less than about 1 g/cm.'}2. The pharmaceutically active article of claim 1 , wherein the pharmaceutically active agent is present at least about 30 wt %.310-. (canceled)11. The pharmaceutically active article of claim 1 , wherein the foam has a specific surface area of at least about 0.5 m/g.1217-. (canceled)18. The pharmaceutically active article of claim 1 , wherein the foam has an average cell size of less than about 4 micrometers.1922-. (canceled)23. The pharmaceutically ...

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Номер записи: 9
15-08-2013 дата публикации

FOAMS, INCLUDING MICROCELLULAR FOAMS, CONTAINING COLLOIDAL PARTICULATES

Номер: US20130209520A1
Автор: Guerra Rodrigo, Kaz David, Koltzenburg Roland S., Ladavac Kosta, Manoharan Vinothan N., Rieger Jens B., Weitz David A.
Принадлежит: BASF SE

The present invention generally relates to foams and particles made from such foams, for applications such as drug delivery. The foams or particles may comprise a pharmaceutically acceptable polymeric carrier. In some cases, the foams may include colloidal particulates. A first aspect of the present invention is generally related to polymer-based foams or particles containing pharmaceutically active agents. In some cases, the foam or particle may contain smaller colloidal particulates therein. Such colloidal particulates may be used, for example, to limit the amount of material within certain regions of the foam, or exclude pharmaceutically active agents from being located within certain portions of the foam, which may useful for enhancing release of pharmaceutically active agents from the foam. In some cases, the colloidal particulates may cause the foam or particle to have an unexpectedly high specific surface area. The foam, in certain embodiments, can exhibit a relatively high loading of the pharmaceutically active agent. The foam may be microcellular in certain instances. The foam may also be created using a supercritical fluid, for example, supercritical C0. For instance, a precursor to the foam, containing a pharmaceutically active agent, a pharmaceutically acceptable polymeric carrier, and colloidal particulates, can be mixed with a foaming agent. The pressure may then be decreased, thereby causing the foaming agent to expand and causing a foam to form. The foam may also be ground or milled, or otherwise processed, to form particles such as nanoparticles. 1. A pharmaceutically active article , comprising:{'sup': '3', 'a foam comprising a pharmaceutically acceptable polymeric carrier and colloidal particulates, wherein (a) the colloidal particulates are present at a density of at least about 1 colloidal particulate/micrometerwithin the foam, (b) the colloidal particulates are present within the foam at a concentration of at least about 20% based on the weight ...

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Номер записи: 10
22-08-2013 дата публикации

Acoustic waves in microfluidics

Номер: US20130213488A1
Автор: Achim Wixforth, Adam R. Abate, David A. Weitz, Jeremy Agresti, Lothar Schmid, Thomas Franke
Принадлежит: Harvard College

Various aspects of the present invention relate to the control and manipulation of fluidic species, for example, in microfluidic systems. In one set of embodiments, droplets may be sorted using surface acoustic waves. The droplets may contain cells or other species. In some cases, the surface acoustic waves may be created using a surface acoustic wave generator such as an interdigitated transducer, and/or a material such as a piezoelectric substrate. The piezoelectric substrate may be isolated from the microfluidic substrate except at or proximate the location where the droplets are sorted, e.g., into first or second microfluidic channels. At such locations, the microfluidic substrate may be coupled to the piezoelectric substrate (or other material) by one or more coupling regions. In some cases, relatively high sorting rates may be achieved, e.g., at rates of at least about 1,000 Hz, at least about 10,000 Hz, or at least about 100,000 Hz, and in some embodiments, with high cell viability after sorting.

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Номер записи: 11
30-01-2014 дата публикации

SYSTEMS AND METHODS FOR SPLITTING DROPLETS

Номер: US20140026968A1
Автор: Abate Adam R., Weitz David A.
Принадлежит:

The present invention generally relates to fluidics and microfluidics and, in particular, to creating droplets in a fluidic system. In some aspects, the present invention is generally directed to systems and methods for splitting a parent droplet into two or more droplets, e.g., by urging the parent droplet towards an obstacle to split the parent droplet. In some cases, the parent droplet is split into at least first and second droplets which each are directed to separate channels. In some cases, the channels may be constructed and arranged such that the droplet velocities of the first and second droplets are substantially the same as the velocity of the parent droplet. In some cases, such droplets may be repeatedly split, e.g., a parent droplet is divided into 2 daughter droplets, then each droplet split again, etc., for example, such that one parent droplet may eventually be split into 2, 2, 2, 2, 2, etc. daughter droplets. In some cases, the daughter droplets may be substantially monodisperse. 1. A method of splitting a parent droplet into two or more droplets , the method comprising:providing a parent droplet flowing at an initial velocity in an inlet microfluidic channel;splitting the parent droplet into at least a first droplet and a second droplet;urging the first droplet into a first microfluidic channel and the second droplet into a second microfluidic channel, the first droplet flowing at a first velocity within the first microfluidic channel and the second droplet flowing at a second velocity within the second microfluidic channel, wherein the first velocity and the second velocity can be the same or different, andwherein the difference in velocities between the fastest and slowest of the initial, first, and second velocities is no more than about 40% of the initial velocity.2. The method of claim 1 , wherein splitting the parent droplet into at least a first droplet and a second droplet comprises urging a first portion of the parent droplet into the ...

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Номер записи: 12
06-02-2014 дата публикации

METHOD AND APPARATUS FOR FLUID DISPERSION

Номер: US20140037514A1
Автор: Anna Shelly L., Bontoux Nathalie, DiLuzio Willow R., Garstecki Piotr, Gitlin Irina, Kumacheva Eugenia, Link Darren Roy, Stone Howard A., Weitz David A., Whitesides George M.
Принадлежит:

A microfluidic method and device for focusing and/or forming discontinuous sections of similar or dissimilar size in a fluid is provided. The device can be fabricated simply from readily-available, inexpensive material using simple techniques. 1. A device comprising:a microfluidic interconnected region comprising an upstream portion, a downstream portion and a dimensionally-restricted section defining the downstream portion of the microfluidic interconnected region, the microfluidic interconnected region being connected at its upstream portion to two or more microfluidic channels.2. The device of claim 1 , wherein the dimensionally-restricted section comprises a non-valved orifice.3. The device of claim 1 , wherein the dimensionally-restricted section is formed by at least an extension of a wall defining the interconnected region.4. The device of claim 1 , wherein the microfluidic interconnected region and the two or more microfluidic channels are part of a single integral unit.5. The device of claim 1 , wherein the microfluidic interconnected region claim 1 , the upstream portion claim 1 , and the downstream portion are each contained within a microfluidic device.6. The device of claim 1 , wherein the microfluidic interconnected region has a maximum cross-sectional diameter of less than 50 microns.7. The device of claim 1 , wherein the downstream portion has a largest dimension perpendicular to fluid flow of less than about 1 mm.8. The device of claim 1 , wherein the device comprises two microfluidic inlet channels each containing a continuous fluid and one microfluidic inlet channel containing a subject fluid.9. The device of claim 8 , wherein the subject fluid forms discontinuous sections at the interconnected region surrounded by the continuous fluid claim 8 , at least some of the discontinuous sections having a maximum dimension of less than 100 microns.10. The device of claim 8 , wherein the continuous fluid completely circumferentially surrounds the subject ...

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Номер записи: 13
06-03-2014 дата публикации

Polymersomes, liposomes, and other species associated with fluidic droplets

Номер: US20140065234A1
Автор: Daeyeon Lee, David A. Weitz, Ho Cheung Shum, Insun Yoon, Jin-woong Kim
Принадлежит: Harvard College

The present invention relates generally to vesicles such as liposomes, colloidosomes, and polymersomes, as well as techniques for making and using such vesicles. In some cases, the vesicles may be at least partially biocompatible and/or biodegradable. The vesicles may be formed, according to one aspect, by forming a multiple emulsion comprising a first droplet surrounded by a second droplet, which in turn is surrounded by a third fluid, where the second droplet comprises lipids and/or polymers, and removing fluid from the second droplet, e.g., through evaporation or diffusion, until a vesicle is formed. In certain aspects, the size of the vesicle may be controlled, e.g., through osmolarity, and in certain embodiments, the vesicle may be ruptured through a change in osmolarity. In some cases, the vesicle may contain other species, such as fluorescent molecules, microparticles, pharmaceutical agents, etc., which may be released upon rupture. Yet other aspects of the invention are generally directed to methods of making such vesicles, kits involving such vesicles, or the like.

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Номер записи: 14
07-01-2016 дата публикации

NANOSTRUCTURED ACTIVE THERAPEUTIC VEHICLES AND USES THEREOF

Номер: US20160000886A1
Автор: Abbaspourrad Alireza, Carroll Nichlaus J., Lind Johan Ulrik, Parker Kevin Kit, Weitz David A.
Принадлежит:

The present invention provides nano structured active therapeutic vehicles which include a biodegradable polymeric fiber and/or thread comprising a porous particle which encapsulates an active agent. The vehicles of the present invention may be used to provide sustained release of the active agent to a subject. 1. A nanostructured active therapeutic vehicle , comprisinga biodegradable polymeric fiber comprising a porous particle or a biodegradable polymeric thread comprising a porous particle, wherein the porous particle comprises regulators that control passage of molecules into and out of the particle, and wherein the porous particle comprises an active agent.2. A nanostructured active therapeutic vehicle for sustained delivery of an active agent , comprising a biodegradable polymeric fiber or a biodegradable polymeric thread and a polymerosome comprising the active agent , wherein the active agent is an agent which inhibits the activity of a toxin , and wherein the polymerosome comprises size regulators which control passage of molecules into and out of the particle such that the active agent is excluded from exiting the polymerosome , a molecule which degrades the active agent is excluded from entry into the polymerosome , and the toxin is permitted entry into the polymerosome such that the toxin contacts the active agent , thereby inhibiting the activity of the toxin.3. The nanostructured active therapeutic vehicle of or , wherein the biodegradable polymeric fiber or biodegradable polymeric thread comprises synthetic and/or natural polymers.4. (canceled)5. (canceled)6. The nanostructured active therapeutic vehicle of or , wherein the polymeric fiber or biodegradable polymeric thread is about 1 to about 1 ,000 micrometers in diameter or about 10 to about 100 micrometers in diameter.7. (canceled)8. The nanostructured active therapeutic vehicle of or , wherein the polymeric fiber or biodegradable polymeric thread has a tensile strength of about 0.5 N to about 100 ...

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Номер записи: 15
02-01-2020 дата публикации

ASSAYS AND OTHER REACTIONS INVOLVING DROPLETS

Номер: US20200002741A1
Автор: Agresti Jeremy, Chu Liang-yin, Church George M., DANTAS Gautam, KIM Jin-Woong, Rowat Amy, SOMMER Morten, Weitz David A.
Принадлежит: President and Fellows of Harvard College

The present invention generally relates to droplets and/or emulsions, such as multiple emulsions. In some cases, the droplets and/or emulsions may be used in assays, and in certain embodiments, the droplet or emulsion may be hardened to form a gel. In some aspects, a heterogeneous assay can be performed using a gel. For example, a droplet may be hardened to form a gel, where the droplet contains a cell, DNA, or other suitable species. The gel may be exposed to a reactant, and the reactant may interact with the gel and/or with the cell, DNA, etc., in some fashion. For example, the reactant may diffuse through the gel, or the hardened particle may liquefy to form a liquid state, allowing the reactant to interact with the cell. As a specific example, DNA contained within a gel particle may be subjected to PCR (polymerase chain reaction) amplification, e.g., by using PCR primers able to bind to the gel as it forms. As the DNA is amplified using PCR, some of the DNA will be bound to the gel via the PCR primer. After the PCR reaction, unbound DNA may be removed from the gel, e.g., via diffusion or washing. Thus, a gel particle having bound DNA may be formed in one embodiment of the invention. 120-. (canceled)21. A composition , comprising:a collection of hardened droplets comprising a library of distinguishable nucleic acid species, wherein a hardened droplet of said collection of hardened droplets comprises a nucleic acid species distinguishable from other nucleic acid species in other hardened droplets of said collection of hardened droplets, and wherein said collection of hardened droplets is contained within a plurality of aqueous droplets in a water-in-oil emulsion.22. The composition of claim 21 , wherein said collection of hardened droplets is a collection of gel particles23. The composition of claim 22 , wherein said collection of gel particles comprises a polyacrylamide polymer.24. The composition of claim 23 , wherein said polyacrylamide polymer comprises a ...

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Номер записи: 16
14-01-2016 дата публикации

DEVICES AND METHODS FOR FORMING RELATIVELY MONODISPERSE DROPLETS

Номер: US20160008778A1
Автор: Amstad Esther, Sperling Ralph Alexander, Weitz David A.
Принадлежит:

Devices and methods for dividing droplets are generally described. In some embodiments, an article may comprise a fluidic channel comprising an array of obstructions. In certain embodiments, the arrangement of obstructions in the array may affect the flow path of fluid in the channel. For example, the array of obstructions may be used to convert a polydisperse population of droplets into a relatively monodisperse population of droplets. Passing a polydisperse population of droplets through the array may result in the division of droplets such that the population of droplets exiting the array has a narrower distribution in the characteristic dimensions of the droplets. The arrangement of obstructions in the array may allow for high-throughput production of a substantially monodisperse population of droplets in some cases. In some embodiments, the population of droplets exiting the array may be converted into particles. 1. An article , comprising:a microfluidic channel comprising a two-dimensional array of obstructions therein, arranged in a plurality of rows of substantially regularly-spaced obstructions, the rows arranged substantially orthogonal to a direction of average fluid flow through the microfluidic channel,wherein at least some of the rows of substantially regularly-spaced obstructions are offset relative to an adjacent row of substantially regularly-spaced obstructions.2. The article of claim 1 , wherein the average horizontal spacing between an obstruction and a next nearest obstruction in the rows of the array is greater than or equal to about 10 micrometers and less than about 100 micrometers.3. The article of any one of or claim 1 , wherein the average vertical spacing between an obstruction and a next nearest obstruction in the columns of the array is greater than or equal to about 10 micrometers and less than about 100 micrometers.43. The article of any one of - claims 1 , wherein the centers of the obstructions in at least some of the rows are ...

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Номер записи: 17
09-01-2020 дата публикации

MANIPULATION OF FLUIDS, FLUID COMPONENTS AND REACTIONS IN MICROFLUIDIC SYSTEMS

Номер: US20200009570A1
Автор: Agresti Jeremy, Boukellal Hakim, Fraden Seth, Jia Yanwei, Rowat Amy, Selimovic Seila, Weitz David A.
Принадлежит:

Microfluidic structures and methods for manipulating fluids, fluid components, and reactions are provided. In one aspect, such structures and methods can allow production of droplets of a precise volume, which can be stored/maintained at precise regions of the device. In another aspect, microfluidic structures and methods described herein are designed for containing and positioning components in an arrangement such that the components can be manipulated and then tracked even after manipulation. For example, cells may be constrained in an arrangement in microfluidic structures described herein to facilitate tracking during their growth and/or after they multiply. 120-. (canceled)21. A method for partitioning a fluid sample , the method comprising:providing a microfluidic device having a substrate comprising a plurality of microwells in fluid communication with an inlet;introducing a first fluid into the inlet of the microfluidic device, the first fluid comprising a biological sample and a plurality of beads, each bead comprising a reactive component for binding a target molecule from the biological sample to the bead; a subvolume of the first fluid,', 'a portion of the biological sample, and', 'more than one bead; and, 'introducing a second fluid immiscible with the first fluid into the microfluidic device and flowing the second fluid towards each of the plurality of microwells so as to form partitions of fluid in corresponding microwells, wherein at least some of the partitions include'}maintaining each partition of fluid in corresponding microwells for detection of contents in each partition of fluid.22. The method of claim 21 , wherein a subset of the partitions do not include any beads and a second subset of the partitions each include a single bead.23. The method of claim 21 , wherein each partition of fluid is separated from the others by the second fluid.24. The method of claim 21 , wherein the second fluid is an oil.25. The method of claim 21 , further ...

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Номер записи: 18
18-01-2018 дата публикации

SYSTEMS, METHODS, AND KITS FOR AMPLIFYING OR CLONING WITHIN DROPLETS

Номер: US20180016622A1
Автор: Heyman John, Mazutis Linas, Weitz David A., Zhang Huidan
Принадлежит: President and Fellows of Harvard College

The present invention generally relates to droplet-based microfluidic devices, including systems, methods, and kits for amplifying or cloning within droplets. In some embodiments, the present invention is generally directed to systems, methods, or kits for amplifying a plurality of nucleic acids, e.g., without substantially selectively amplifying some nucleic acids over others. The nucleic acids may be contained within the droplets. In addition, in some embodiments, a plurality of microfluidic droplet containing a species of interest, such as a nucleic acid, may be mixed with microfluidic droplets free of the species, then pipetted or otherwise transferred such that, on average, a predetermined number of droplets containing species of interest is transferred. 1. A method , comprising:fragmenting a nucleic acid to produce nucleic acid fragments;containing at least some of the nucleic acid fragments in a plurality of microfluidic droplets; andamplifying at least some of the nucleic acid fragments contained within the microfluidic droplets.2. The method of claim 1 , comprising using PCR to amplifying at least some of the nucleic acid fragments contained within the microfluidic droplets.3. The method of any one of or claim 1 , wherein amplifying at least some of the nucleic acid fragments comprises adding a polymerase to at least some of the microfluidic droplets.4. The method of claim 3 , wherein the polymerase is DNA polymerase.5. The method of claim 3 , wherein the polymerase is RNA polymerase.6. The method of any one of - claim 3 , wherein amplifying at least some of the nucleic acid fragments comprises adding Taq polymerase to at least some of the microfluidic droplets.7. The method of any one of - claim 3 , wherein at least some of the microfluidic droplets comprise Taq polymerase.8. The method of any one of - claim 3 , comprising exposing at least some of the microfluidic droplets to a temperature of at least about 50° C.9. The method of any one of - claim 3 , ...

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Номер записи: 19
28-01-2016 дата публикации

SPRAY DRYING TECHNIQUES

Номер: US20160023126A1
Автор: Abate Adam R., HOLTZE Christian, Thiele Julian W.P., Weitz David A., Windbergs Maike
Принадлежит:

The present invention generally relates to microfluidics, and to spray drying and other drying techniques. In some aspects, an article containing one or more channels or microfluidic channels is used to mix one or more fluids prior to spray drying. The mixing may occur immediately before the fluids are expelled through a nozzle or other opening into a drying region of the spray dryer. In one set of embodiments, for example, a first fluid is exposed to a second fluid, then the fluids are exposed to air or other gases before being expelled through a nozzle. In certain instances, the first fluid may contain a dissolved species that may precipitate upon exposure to the second fluid; such precipitation may occur immediately before expulsion through a nozzle or other opening, thereby resulting in controlled precipitation as part of the spray drying process. 183-. (canceled)84. A method of spray drying , the method comprising:flowing a first liquid through a first microfluidic channel having an opening as a nozzle, wherein the first liquid comprises a first species dissolved in the first liquid;flowing a second liquid through a second microfluidic channel intersecting the first microfluidic channel at an intersection upstream of the nozzle;exposing the first liquid to the second liquid within the first microfluidic channel, wherein the first liquid is surrounded by the second liquid downstream of the intersection such that the first liquid does not contact a wall of the first microfluidic channel proximate to the nozzle, wherein the first species precipitates upon exposure of the first liquid to the second liquid; andspraying the first liquid and the second liquid into a drying region external of the first and second microfluidic channels.85. The method of claim 84 , wherein the first liquid and the second liquid are immiscible.86. The method of claim 84 , further comprising flowing the second liquid through a third microfluidic channel intersecting the first microfluidic ...

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Номер записи: 20
25-01-2018 дата публикации

SYSTEM AND METHOD FOR A BIOMIMETIC FLUID PROCESSING

Номер: US20180021780A1
Автор: Italiano Joseph, Mazutis Linas, Thon Jonathan N., Weitz David A.
Принадлежит:

A system and method are provided for harvesting target biological substances. The system includes a substrate and a first and second channel formed in the substrate. The channels longitudinally extending substantially parallel to each other. A series of gaps extend from the first channel to the second channel to create a fluid communication path passing between a series of columns with the columns being longitudinally separated by a predetermined separation distance. The system also includes a first source configured to selectively introduce into the first channel a first biological composition at a first channel flow rate and a second source configured to selectively introduce into the second channel a second biological composition at a second channel flow rate. The sources are configured to create a differential between the first and second channel flow rates to generate physiological shear rates along the second channel that are bounded within a predetermined range. 1. A biomimetic microfluidic system comprising:a substrate having a first portion and a second portion;a first channel formed in the first portion of the substrate, the first channel extending from a first input to a first output along a longitudinal dimension, the first channel configured to selectively receive at least one first biological composition at a first channel flow rate, the at least one first biological composition including a biological source material;a second channel formed in the second portion of the substrate, the second channel extending from a second input to a second output along the longitudinal dimension, the second channel configured to selectively receive at least one second biological composition at a second channel flow rate, wherein at least a portion of the first and second channels are substantially parallel;microchannels extending between the first channel and the second channel to create a fluid communication path between the first and second channels, wherein the ...

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Номер записи: 21
25-01-2018 дата публикации

ASSAY AND OTHER REACTIONS INVOLVING DROPLETS

Номер: US20180023109A1
Автор: Agresti Jeremy, Chu Liang-yin, Church George, DANTAS Gautam, KIM Jin-Woong, Rowat Amy, SOMMER Morten, Weitz David A.
Принадлежит:

The present invention generally relates to droplets and/or emulsions, such as multiple emulsions. In some cases, the droplets and/or emulsions may be used in assays, and in certain embodiments, the droplet or emulsion may be hardened to form a gel. In some aspects, a heterogeneous assay can be performed using a gel. For example, a droplet may be hardened to form a gel, where the droplet contains a cell, DNA, or other suitable species. The gel may be exposed to a reactant, and the reactant may interact with the gel and/or with the cell, DNA, etc., in some fashion. For example, the reactant may diffuse through the gel, or the hardened particle may liquefy to form a liquid state, allowing the reactant to interact with the cell. As a specific example, DNA contained within a gel particle may be subjected to PCR (polymerase chain reaction) amplification, e.g., by using PCR primers able to bind to the gel as it forms. As the DNA is amplified using PCR, some of the DNA will be bound to the gel via the PCR primer. After the PCR reaction, unbound DNA may be removed from the gel, e.g., via diffusion or washing. Thus, a gel particle having bound DNA may be formed in one embodiment of the invention. 120-. (canceled)21. A method , comprising:providing a gel droplet containing a species, wherein the gel droplet is within a surrounding droplet; andexposing the species within the gel droplet to a reactant which is reactive with the species.22. The method of claim 21 , wherein providing a gel droplet containing a species comprises:providing a fluidic droplet containing a species; andcausing the fluidic droplet to form a gel droplet containing the species, wherein the gel droplet is within a surrounding droplet.23. The method of claim 21 , wherein exposing the species occurs after releasing the species from the gel droplet.24. The method of claim 21 , wherein the species is a first nucleic acid claim 21 , siRNA claim 21 , RNAi or DNA.25. The method of claim 24 , wherein the reactant ...

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Номер записи: 22
25-01-2018 дата публикации

SYSTEMS AND METHODS FOR EPIGENETIC SEQUENCING

Номер: US20180023133A1
Автор: BERNSTEIN BRADLEY E., Ram Oren, Rotem Assaf, Weitz David A.
Принадлежит:

The present invention generally relates to microfluidics and/or epigenetic sequencing. In one set of embodiments, cells contained within a plurality of microfluidic droplets are lysed and the DNA (e.g., from nucleosomes) within the droplets are labeled, e.g., with adapters containing an identification sequence. The adapters may also contain other sequences, e.g., restriction sites, primer sites, etc., to assist with later analysis. After labeling with adapters, the DNA from the different cells may be combined and analyzed, e.g., to determine epigenetic information about the cells. For example, the DNA may be separated on the basis of certain modifications (e.g., methylation), and the DNA from the separated nucleosomes may be sequenced using techniques such as chromatin immunoprecipitation (“ChIP”). In some cases, the DNA sequences may also be aligned with genomes, e.g., to determine which portions of the genome were epigenetically modified, e.g., via methylation. 1111-. (canceled)112. A method comprising:providing a solution comprising a plurality of nucleic acid sequences originating from a plurality of cells, at least some of the nucleic acid sequences being attached to an adapter, the adapter comprising an identification sequence, wherein sequences originating from the same cell contain identical identification sequences, and sequences originating from different cells contain different identification sequences; and sequencing at least some of the sequences.113. The method of claim 112 , wherein the adapter comprises a restriction site.114. The method of claim 112 , comprising cleaving the adapter at the restriction site.115. The method of claim 112 , wherein the solution is contained within a droplet.116. The method of claim 112 , wherein the plurality of nucleic acid sequences comprise DNA.117. The method of claim 112 , wherein the plurality of nucleic acid sequences comprise RNA.118. The method of claim 112 , wherein the primer site comprises a universal primer ...

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Номер записи: 23
24-01-2019 дата публикации

COMPARTMENTALISED COMBINATORIAL CHEMISTRY BY MICROFLUIDIC CONTROL

Номер: US20190024261A1
Автор: Ahn Keunho, Bibette Jérôme, Griffiths Andrew David, Link Darren Roy, Weitz David A.
Принадлежит:

The invention describes a method for the synthesis of compounds comprising the steps of: (a) compartmentalizing two or more sets of primary compounds into microcapsules; such that a proportion of the microcapsules contains two or more compounds; and (b) forming secondary compounds in the microcapsules by chemical reaction between primary compounds from different sets; wherein one or both of steps (a) and (b) is performed under microfluidic control; preferably electronic microfluidic control. The invention further allows for the identification of compounds which bind to a target component of a biochemical system or modulate the activity of the target, and which is co-compartmentalized into the microcapsules. 1. A method for analyzing compounds , the method comprising:forming a plurality of microcapsules, wherein each microcapsule comprises a microbead and a cell, and wherein each microbead comprises one of a plurality of compounds;performing a reaction within each microcapsule, wherein the reaction involves the compound and a molecule associated with the cell;pooling the plurality of microcapsules into a common compartment; andidentifying the microbeads comprising a portion of the compounds that cause a selectable change in a product of the reaction.2. The method of claim 1 , wherein each of the plurality of microcapsules is surrounded by an immiscible fluid comprising a surfactant.3. The method of claim 1 , wherein the plurality of microcapsules are monodisperse.4. The method of claim 1 , wherein each of the plurality of microcapsules comprises no more than one microbead.5. The method of claim 1 , wherein the plurality of compounds comprise nucleic acids or proteins.6. The method of claim 1 , wherein the plurality of compounds are labeled.7. The method of claim 1 , wherein each of the plurality of compounds comprises a primer sequence for amplification.8. The method of claim 1 , wherein the reaction is an amplification reaction.9. The method of claim 1 , wherein the ...

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Номер записи: 24
02-02-2017 дата публикации

SYSTEMS AND METHODS FOR DROPLET TAGGING

Номер: US20170028377A1
Автор: BERNSTEIN BRADLEY E., Nicol Robert, Weitz David A.
Принадлежит:

The present invention generally relates to microfluidic devices, including systems and methods for tagging droplets within such devices. In some aspects, microfluidic droplets are manipulated by exposing the droplets (or other discrete entities) to a variety of different conditions. By incorporating into the droplets a plurality of nucleic acid “tags,” and optionally ligating then nucleic acids together, the conditions that a droplet was exposed to may be encoded by the nucleic acid tags. Thus, even if droplets exposed to different conditions are mixed together, the conditions that each droplet encountered may still be determined, for example, by sequencing the nucleic acids. 1. A method , comprising:exposing a droplet to a first condition and adding a first nucleic acid to the droplet, wherein the first nucleic acid encodes the first condition;exposing the droplet to a second condition and adding a second nucleic acid to the droplet, wherein the second nucleic acid encodes the second condition; andligating the first nucleic acid and the second nucleic acid together.2. The method of claim 1 , wherein exposing the droplet to the first condition and adding the first nucleic acid to the droplet comprises fusing the droplet to a second droplet containing the first nucleic acid.3. The method of any one of or claim 1 , wherein exposing the droplet to the first condition comprises exposing the droplet to a molecular species.4. The method of claim 3 , wherein exposing the droplet to the molecular species comprises fusing the droplet with a second droplet containing the molecular species.5. The method of claim 4 , wherein the second droplet further contains the first nucleic acid claim 4 , whereby when the second droplet is fused to the droplet claim 4 , the first nucleic acid is added to the droplet.6. The method of any one of or claim 4 , wherein the second droplet is fused to the droplet via dipoles induced in the droplets.7. The method of any one of or claim 4 , wherein ...

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Номер записи: 25
02-02-2017 дата публикации

METHODS AND SYSTEMS FOR DROPLET TAGGING AND AMPLIFICATION

Номер: US20170029813A1
Автор: Iafrate Anthony John, Weitz David A., Zhang Huidan, Zheng Zongli
Принадлежит:

The present invention generally relates to microfluidic devices, including methods and systems for tagging droplets within such devices. In some aspects, microfiuidic droplets are manipulated by exposing the droplets (or other discrete entities) to a variety of different conditions. By incorporating into the droplets a plurality of nucleic acid “tags,” and optionally amplifying the nucleic acids, e.g., within the droplets, the conditions that a droplet was exposed to may be encoded by the nucleic acids. Thus, even if droplets exposed to different conditions are mixed together, the conditions that each droplet encountered may still be determined, for example, by sequencing the nucleic acids. 1. A method , comprising:exposing a microfluidic droplet to a first condition and adding a first nucleic acid to the droplet, wherein the first nucleic acid encodes the first condition;exposing the microfluidic droplet to a second condition and adding a second nucleic acid to the droplet, wherein the second nucleic acid encodes the second condition; andamplifying the first nucleic acid and the second nucleic acid within the microfluidic droplet.2. The method of claim 1 , wherein exposing the microfluidic droplet to the first condition and adding the first nucleic acid to the droplet comprises fusing the microfluidic droplet to a second droplet containing the first nucleic acid.3. The method of any one of or claim 1 , wherein exposing the microfluidic droplet to the first condition comprises exposing the droplet to a molecular species.4. The method of claim 3 , wherein exposing the microfluidic droplet to the molecular species comprises fusing the microfluidic droplet with a second droplet containing the molecular species.5. The method of claim 4 , wherein the second droplet further contains the first nucleic acid claim 4 , whereby when the second droplet is fused to the microfluidic droplet claim 4 , the first nucleic acid is added to the microfluidic droplet.6. The method of any ...

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Номер записи: 26
01-02-2018 дата публикации

Droplet-Based Method And Apparatus For Composite Single-Cell Nucleic Acid Analysis

Номер: US20180030515A1
Автор: BASU Anindita, FORD Christopher B., MACOSKO Evan Zane, McCarroll Steven Andrew, Park Hongkun, Regev Aviv, Shalek Alexander k., Weitz David A.
Принадлежит:

The present invention generally relates to a combination of molecular barcoding and emulsion-based microfluidics to isolate, lyse, barcode, and prepare nucleic acids from individual cells in a high-throughput manner. 197-. (canceled)98. A nucleotide- or oligonucleotide-adorned bead wherein said bead comprises:(a) a linker;(b) an identical sequence for use as a sequencing priming site;(c) a uniform or near-uniform nucleotide or oligonucleotide sequence;(d) a Unique Molecular Identifier which differs for each priming site;(e) optionally an oligonucleotide redudant sequence for capturing polyadenylated mRNAs and priming reverse transcription; and(f) optionally at least one other oligonucleotide barcode which provides an additional substrate for identification.99. The nucleotide- or oligonucleotide-adorned bead of claim 98 , wherein:the nucleotide or oligonucleotide sequence on the surface of the bead is a molecular barcode; orthe nucleotide or oligonucleotide sequence on the surface of the bead is a molecular barcode from 4 to 1000 nucleotides in length; orthe oligonucleotide sequence for capturing polyadenylated mRNAs and priming reverse transcription is an oligo dT sequence; orthe linker is a non-cleavable, straight-chain polymer; orthe linker is a chemically-cleavable, straight-chain polymer; orthe linker is a non-cleavable optionally substituted hydrocarbon polymer; orthe linker is a photolabile optionally substituted hydrocarbon polymer; orthe linker is a polyethylene glycol; or{'sub': '3-24', 'the linker is a PEG-C.'}100. A mixture comprising a plurality of nucleotide- or oligonucleotide- adorned beads claim 98 , wherein said beads comprises;(a) a linker;(b) an identical sequence for use as a sequencing priming site;(c) a uniform or near-uniform nucleotide or oligonucleotide sequence;(d) a Unique Molecular Identifier which differs for each priming site;(e) an oligonucleotide redudant sequence for capturing polyadenylated mRNAs and priming reverse transcription; ...

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Номер записи: 27
05-02-2015 дата публикации

DROPLET FORMATION USING FLUID BREAKUP

Номер: US20150034163A1
Автор: Abate Adam R., Weitz David A.
Принадлежит:

The present invention generally relates to systems and methods for creating droplets. In one aspect, a plurality of droplets () is introduced into a continuous fluid stream () to cause the continuous fluid stream to form discrete droplets. In some cases, the droplets that are formed from the continuous fluid stream may be substantially monodisperse. The continuous fluid stream may, in some cases, be a jetting fluid stream flowing at a relatively high linear flow rate, and in certain embodiments, high rates of droplet formation from the jetting fluid may thereby be achieved. Additionally, certain aspects of the invention are generally directed to devices, such as microfluidic devices, able to form such droplets. For example, in one set of embodiments, a device may include a junction () where a plurality of droplets () can be introduced into a continuous fluid stream (), and optionally, the device may include additional junctions () able to cause the formation of the plurality of droplets and/or the formation of the continuous fluid stream. Still other disclosed aspects are generally directed to methods of making such devices, methods of using such devices, kits involving such devices, and the like. 1. A method of producing droplets , comprising:providing, in a microfluidic channel, a continuous fluid stream comprising a first fluid; andinserting a plurality of droplets of a second fluid into the continuous fluid stream to cause the continuous fluid stream to form discrete droplets of first fluid.2. The method of claim 1 , wherein the continuous fluid stream comprising the first fluid is a fluid jet.3. The method of claim 1 , wherein the continuous fluid stream comprising the first fluid has a flow rate such that claim 1 , in the absence of the insertion of the plurality of droplets of second fluid claim 1 , the continuous fluid stream does not form discrete droplets of first fluid.4. The method of claim 1 , wherein the continuous fluid stream comprising the first ...

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Номер записи: 28
07-02-2019 дата публикации

MANIPULATION OF FLUIDS, FLUID COMPONENTS AND REACTIONS IN MICROFLUIDIC SYSTEMS

Номер: US20190039071A1
Автор: Agresti Jeremy, Boukellal Hakim, Fraden Seth, Jia Yanwei, Rowat Amy, Selimovic Seila, Weitz David A.
Принадлежит:

Microfluidic structures and methods for manipulating fluids, fluid components, and reactions are provided. In one aspect, such structures and methods can allow production of droplets of a precise volume, which can be stored/maintained at precise regions of the device. In another aspect, microfluidic structures and methods described herein are designed for containing and positioning components in an arrangement such that the components can be manipulated and then tracked even after manipulation. For example, cells may be constrained in an arrangement in microfluidic structures described herein to facilitate tracking during their growth and/or after they multiply. 179.-. (canceled)80. A method for partitioning a fluid sample , the method comprising:providing a microfluidic device having a substrate comprising a plurality of microwells in fluid communication with an inlet;introducing a first fluid into the inlet of the microfluidic device, the first fluid comprising a biological sample and a plurality of beads, each bead comprising a reactive component for binding a target molecule from the biological sample to the bead;introducing a second fluid immiscible with the first fluid into the microfluidic device and flowing the second fluid towards each of the plurality of microwells so as to form partitions of fluid in corresponding microwells, each partition of fluid comprising a subvolume of the first fluid, including at least the biological sample and a single bead, wherein each partition of fluid is separated from one another by the second fluid; andmaintaining each partition of fluid in corresponding microwells for detection of contents in each partition of fluid.81. The method of claim 80 , wherein each partition of fluid is at least partially surrounded by the second fluid.82. The method of claim 80 , wherein the second fluid is an oil.83. The method of claim 80 , further comprising monitoring each partition of fluid and detecting claim 80 , with a detector claim 80 ...

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Номер записи: 29
26-02-2015 дата публикации

SYSTEMS AND METHODS FOR EPIGENETIC SEQUENCING

Номер: US20150057163A1
Автор: BERNSTEIN BRADLEY E., Ram Oren, Rotem Assaf, Weitz David A.
Принадлежит:

The present invention generally relates to microfluidics and/or epigenetic sequencing. In one set of embodiments, cells contained within a plurality of microfluidic droplets are lysed and the DNA (e.g., from nucleosomes) within the droplets are labeled, e.g., with adapters containing an identification sequence. The adapters may also contain other sequences, e.g., restriction sites, primer sites, etc., to assist with later analysis. After labeling with adapters, the DNA from the different cells may be combined and analyzed, e.g., to determine epigenetic information about the cells. For example, the DNA may be separated on the basis of certain modifications (e.g., methylation), and the DNA from the separated nucleosomes may be sequenced using techniques such as chromatin immunoprecipitation (“ChIP”). In some cases, the DNA sequences may also be aligned with genomes, e.g., to determine which portions of the genome were epigenetically modified, e.g., via methylation. 188-. (canceled)89. A method , comprising:providing a solution comprising a plurality of sequences originating from a plurality of cells, at least some of the sequences being ligated to an adapter, the adapter comprising an identification sequence and a restriction site, wherein sequences originating from the same cell contain identical identification sequences, and sequences originating from different cells contain different identification sequences; andsequencing at least some of the sequences.90. The method of claim 89 , wherein the plurality of sequences comprise DNA sequences.91. The method of claim 89 , wherein the plurality of sequences comprise RNA sequences.92. The method of claim 89 , wherein the solution is contained within a compartment.93. The method of claim 89 , wherein the solution is contained within a droplet.9499-. (canceled)100. The method of claim 89 , wherein the identification sequences comprise 8 nucleotides.101. The method of claim 89 , wherein a plurality of adapters are used claim ...

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Номер записи: 30
01-03-2018 дата публикации

Droplet creation techniques

Номер: US20180056293A1
Автор: Adam R. Abate, David A. Weitz
Принадлежит: Harvard College

The present invention is generally related to systems and methods for producing droplets. The droplets may contain varying species, e.g., for use as a library. In some cases, at least one droplet is used to create a plurality of droplets, using techniques such as flow-focusing techniques. In one set of embodiments, a plurality of droplets, containing varying species, can be divided to form a collection of droplets containing the various species therein. A collection of droplets, according to certain embodiments, may contain various subpopulations of droplets that all contain the same species therein. Such a collection of droplets may be used as a library in some cases, or may be used for other purposes.

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Номер записи: 31
01-03-2018 дата публикации

SYSTEMS AND METHODS FOR NUCLEIC ACID SEQUENCING

Номер: US20180057875A1
Автор: Abate Adam R., Agresti Jeremy, Hung Tony, Weiner Michael P., Weitz David A.
Принадлежит:

The present invention relates to systems and methods for sequencing nucleic acids, including sequencing nucleic acids in fluidic droplets. In one set of embodiments, the method employs sequencing by hybridization using droplets such as microfluidic droplets. In some embodiments, droplets are formed which include a target nucleic acid, a nucleic acid probe, and at least one identification element, such as a fluorescent particle. The nucleic acid probes that hybridize to the target nucleic acid are determined, in some instances, by determining the at least one identification element. The nucleic acid probes that hybridize to the target nucleic acid may be used to determine the sequence of the target nucleic acid. In certain instances, the microfluidic droplets are provided with reagents that modify the nucleic acid probe. In some cases, a droplet, such as those described above, is deformed such that the components of the droplets individually pass a target area. 1109-. (canceled)110. A method , comprising:providing a first microfluidic droplet containing a nucleic acid probe and at least three distinguishable identification elements, wherein each of the at least three distinguishable identification elements is associated with a different oligonucleotide sequence;providing a second microfluidic droplet comprising a target nucleic acid; andfusing at least some of the fluid in the first fluidic droplet and at least some of the fluid in the second microfluidic droplet to form a fused droplet.111. The method of claim 110 , further comprising determining the target nucleic acid contained within the fused droplet.112. The method of claim 110 , comprising determining a sequence of at least a portion of the target nucleic acid.113. The method of claim 110 , further comprising determining at least a portion of the target nucleic acid by determining at least one of the at least three identification elements contained within the fused droplet.114. The method of claim 110 , ...

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Номер записи: 32
08-03-2018 дата публикации

DETERMINATION OF CELLS USING AMPLIFICATION

Номер: US20180066305A1
Автор: Weitz David A., Zhang Huidan
Принадлежит: President and Fellows of Havard College

The present invention generally relates to microfluidics and, in particular, to systems and methods for determining cells using amplification. In one set of embodiments, cells are encapsulated within droplets and nucleic acids from the cells amplified within the droplets. The droplets may then be pooled together and the amplified nucleic acids can be determined using PCR or other suitable techniques. In some embodiments, techniques such as these can be used to detect relatively rare cells that may be present, e.g., if the droplets are amplified using conditions able to selectively amplify nucleic acids arising from the relatively rare cells. 1. A method , comprising:encapsulating cells within a plurality of microfluidic droplets;lysing at least some of the cells within the plurality of droplets to release nucleic acids from the cells into the interior of the droplets;within the interior of the droplets, applying conditions able to selectively amplify a target nucleic acid sequence suspected of being present within the nucleic acids released from the cells;combining the interiors of the droplets together to form a combined fluid; anddetermining amplified nucleic acids contained within the combined fluid.2. The method of claim 1 , wherein the cells comprise human cells.3. The method of any one of or claim 1 , wherein the cells comprise cancer cells.4. The method of any one of - claim 1 , wherein the cells comprise leukemia cells.5. The method of any one of - claim 1 , wherein the cells comprise blood cells.6. The method of any one of - claim 1 , wherein the cells are taken from a biopsy.7. The method of any one of - claim 1 , wherein the cells comprise bacteria.8. The method of any one of - claim 1 , wherein the cells comprise cells suspected of causing a disease.9. The method of any one of - claim 1 , wherein the target nucleic acid sequence is a DNA sequence.10. The method of any one of - claim 1 , wherein the target nucleic acid sequence is an RNA sequence.11. The ...

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Номер записи: 33
05-06-2014 дата публикации

SURFACES, INCLUDING MICROFLUIDIC CHANNELS, WITH CONTROLLED WETTING PROPERTIES

Номер: US20140150916A1
Автор: Abate Adam R., HOLTZE Christian, Krummel Amber T., Weitz David A.
Принадлежит: BASF SE

The present invention generally relates to coating materials, including photoactive coating materials. In some aspects of the invention, a sol-gel is provided that can be formed as a coating on a microfluidic channel. One or more portions of the sol-gel can be reacted to alter its hydrophobicity, in some cases. For instance, in one set of embodiments, a portion of the sol-gel may be exposed to light, such as ultraviolet light, which can be used to induce a chemical reaction in the sol-gel that alters its hydrophobicity. In one set of embodiments, the sol-gel can include a photoinitiator, that upon exposure to light, produces radicals. Optionally, the photoinitiator may be conjugated to a silane or other material within the sol-gel. The radicals so produced may be used to cause a polymerization reaction to occur on the surface of the sol-gel, thus altering the hydrophobicity of the surface. In some cases, various portions may be reacted or left unreacted, e.g., by controlling exposure to light (for instance, using a mask). Such treated surfaces within a microfluidic channel may be useful in a wide variety of applications, for instance, in the creation of emulsions such as multiple emulsions. 1. An article , comprising:a sol-gel coating coated on at least a portion of a microfluidic channel, wherein the sol-gel comprises a moiety containing a double bond.2. The article of claim 1 , wherein the microfluidic channel is defined within a microfluidic device comprising polydimethylsiloxane.3. The article of claim 1 , wherein the microfluidic channel is defined within a microfluidic device comprising glass.4. The article of claim 1 , wherein a first portion of the sol-gel coating is relatively hydrophobic claim 1 , and a second portion of the sol-gel coating is relatively hydrophilic.5. The article of claim 1 , wherein the coating comprises a silane.6. The article of claim 1 , wherein the coating comprises a fluorosilane.7. The article of claim 1 , wherein the coating ...

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Номер записи: 34
15-03-2018 дата публикации

MULTIPLE EMULSIONS CREATED USING JETTING AND OTHER TECHNIQUES

Номер: US20180071695A1
Автор: Abate Adam R., Thiele Julian W.P., Weitz David A.
Принадлежит: President and Fellows of Harvard College

The present invention generally relates to emulsions, and more particularly, to multiple emulsions. In one aspect, multiple emulsions are formed by urging a fluid into a channel, e.g., by causing the fluid to enter the channel as a “jet.” Side channels can be used to encapsulate the fluid with a surrounding fluid. In some cases, multiple fluids may flow through a channel collinearly before multiple emulsion droplets are formed. The fluidic channels may also, in certain embodiments, include varying degrees of hydrophilicity or hydrophobicity. As examples, the fluidic channel may be relatively hydrophilic upstream of an intersection (or other region within the channel) and relatively hydrophobic downstream of the intersection, or vice versa. In some cases, the average cross-sectional dimension may change, e.g., at an intersection. For instance, the average cross-sectional dimension may increase at the intersection. Surprisingly, a relatively small increase in dimension, in combination with a change in hydrophilicity of the fluidic channel, may delay droplet formation of a stream of collinearly-flowing multiple fluids under certain flow conditions; accordingly, the point at which multiple emulsion droplets are formed can be readily controlled within the fluidic channel. In some cases, the multiple droplet may be formed from the collinear flow of fluids at (or near) a single location within the fluidic channel. In addition, unexpectedly, systems such as those described herein may be used to encapsulate fluids in single or multiple emulsions that are difficult or impossible to encapsulate using other techniques, such as fluids with low surface tension, viscous fluids, or viscoelastic fluids. Other aspects of the invention are generally directed to methods of making and using such systems, kits involving such systems, emulsions created using such systems, or the like. 160-. (canceled)61. A method , comprising:providing a first aqueous fluid in a main microfluidic channel; ...

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Номер записи: 35
15-03-2018 дата публикации

SYSTEMS AND METHODS FOR BARCODING NUCLEIC ACIDS

Номер: US20180071705A1
Автор: Akartuna Ilke, Kirschner Marc W., Klein Allon Moshe, Mazutis Linas, Weitz David A.
Принадлежит:

The present invention generally relates to microfluidics and labeled nucleic acids. For example, certain aspects are generally directed to systems and methods for labeling nucleic acids within microfluidic droplets. In one set of embodiments, the nucleic acids may include “barcodes” or unique sequences that can be used to distinguish nucleic acids in a droplet from those in another droplet, for instance, even after the nucleic acids are pooled together. In some cases, the unique sequences may be incorporated into individual droplets using particles and attached to nucleic acids contained within the droplets (for example, released from lysed cells). In some cases, the barcodes may be used to distinguish tens, hundreds, or even thousands of nucleic acids, e.g., arising from different cells or other sources. 1. A method , comprising:i) providing a plurality of at least 10,000 microfluidic droplets containing cells, at least about 90% of the plurality of droplets containing one cell or no cell;ii) lysing the cells within the plurality of microfluidic droplets to release nucleic acid from the cells; and a) wherein the oligonucleotide tag uniquely identifies the released nucleic acids from nucleic acids released from other cells by a barcode sequence and the tag additionally comprises a primer sequence; and', 'b) wherein for at least about 90% of the droplets, the oligonucleotide tag within the droplet is distinguishable from oligonucleotide tags within other droplets of the plurality of droplets., 'iii) bonding the released nucleic acid to oligonucleotide tags,'}2. A method according to claim 1 , wherein at least some of the oligonucleotide tags comprise a poly-T sequence.3. A method according to claim 1 , wherein the nucleic acid is an RNA.4. A method according to claim 1 , wherein the RNA is reverse transcribed within the droplet.5. A method according to claim 4 , wherein after the reverse transcription a barcoded cDNA is created.6. A method according to claim 5 , ...

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Номер записи: 36
31-03-2022 дата публикации

FORMATION AND CONTROL OF FLUIDIC SPECIES

Номер: US20220097067A1
Автор: Cheng Zhengdong, Link Darren Roy, Marquez-Sanchez Manuel, Weitz David A.
Принадлежит: President and Fellows of Harvard College

This invention generally relates to systems and methods for the formation and/or control of fluidic species, and articles produced by such systems and methods. In some cases, the invention involves unique fluid channels, systems, controls, and/or restrictions, and combinations thereof. In certain embodiments, the invention allows fluidic streams (which can be continuous or discontinuous, i.e., droplets) to be formed and/or combined, at a variety of scales, including microfluidic scales. In one set of embodiments, a fluidic stream may be produced from a channel, where a cross-sectional dimension of the fluidic stream is smaller than that of the channel, for example, through the use of structural elements, other fluids, and/or applied external fields, etc. In some cases, a Taylor cone may be produced. In another set of embodiments, a fluidic stream may be manipulated in some fashion, for example, to create tubes (which may be hollow or solid), droplets, nested tubes or droplets, arrays of tubes or droplets, meshes of tubes, etc. In some cases, droplets produced using certain embodiments of the invention may be charged or substantially charged, which may allow their further manipulation, for instance, using applied external fields. Non-limiting examples of such manipulations include producing charged droplets, coalescing droplets (especially at the microscale), synchronizing droplet formation, aligning molecules within the droplet, etc. In some cases, the droplets and/or the fluidic streams may include colloids, cells, therapeutic agents, and the like. 1. A microfluidic device for producing a plurality of droplets comprising:a first microfluidic channel for flowing a first liquid;a second microfluidic channel for flowing a second liquid, the second microfluidic channel meeting the first microfluidic channel at a junction; andelectrodes operably associated with the junction and configured to provide an electric field at the junction to generate droplets of the first ...

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Номер записи: 37
25-03-2021 дата публикации

MICROFLUIDIC DEVICE FOR STORAGE AND WELL-DEFINED ARRANGEMENT OF DROPLETS

Номер: US20210086183A1
Автор: Agresti Jeremy, BOEHM Christian, Koester Sarah, Rowat Amy, Weitz David A.
Принадлежит: President and Fellows of Harvard College

The present invention relates to systems and methods for the arrangement of droplets in pre-determined locations. Many applications require the collection of time-resolved data. Examples include the screening of cells based on their growth characteristics or the observation of enzymatic reactions. The present invention provides a tool and related techniques which addresses this need, and which can be used in many other situations. The invention provides, in one aspect, a tool that allows for stable storage and indexing of individual droplets. The invention can interface not only with microfluidic/microscale equipment, but with macroscopic equipment to allow for the easy injection of liquids and extraction of sample droplets, etc. 171-. (canceled)72. A method of arranging cells in an array , the method comprising:flowing cells through a plurality of storage channels arranged in series, wherein each storage channel comprises one or more pots each comprising a constriction that is smaller than an average cross-sectional dimension of a single cell;trapping a single cell in each of the one or more pots in a first of the plurality of storage channels; andflowing cells around the first of the plurality of storage channels through one or more bypass channels.73. The method of claim 72 , wherein each pot has no more than two constrictions and is in direct fluid communication with no more than two other pots.74. The method of claim 72 , wherein the volumes of the one or more pots are about 10 nanoliters.75. The method of claim 72 , further comprising separating the contents of the one or more pots from a controlled environment by a gas-permeable membrane.76. The method of claim 72 , further comprising separating the contents of the one or more pots from ambient atmosphere by a gas-permeable membrane.77. The method of claim 72 , further comprising performing a reaction on the trapped single cell.78. The method of claim 77 , wherein the one or more pots are associated with a ...

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Номер записи: 38
29-03-2018 дата публикации

BARCODING SYSTEMS AND METHODS FOR GENE SEQUENCING AND OTHER APPLICATIONS

Номер: US20180087078A1
Автор: Heyman Hohn, Klein Allon Moshe, Weitz David A., Zhang Huidan
Принадлежит: President and Fellows of Harvard College

The present invention generally relates to microfluidics and labeled nucleic acids. In one aspect, the present invention is generally directed to a method, wherein the method includes providing a plurality of droplets comprising particles, the particles comprising oligonucleotides, and attaching a nucleic acid sequence to the oligonucleotides. Certain embodiments are generally directed to systems and methods for splitting a droplet into two or more droplets. Certain embodiments are generally directed to systems and methods for sorting fluidic droplets in a liquid. 196-. (canceled)98. The method of claim 97 , wherein the pre-defined pool of first barcodes comprises at least about 300 distinguishable barcodes.99. The method of claim 97 , wherein the pre-defined pool of second barcodes comprises at least about 300 distinguishable barcodes.100. The method of claim 97 , wherein the nucleic acid sequence is configured to bind to genomic DNA.101. The method of claim 97 , wherein at least some of the oligonucleotides are attached to the surface of the particles.102. The method of claim 97 , wherein at least some of the oligonucleotides comprise a cleavable linker.103. The method of claim 102 , wherein the cleavable linker is a photocleavable linker.104. The method of claim 102 , wherein the cleavable linker is a chemically cleavable linker.105. The method of claim 102 , wherein the cleavable linker is an enzymatically cleavable linker.106. The method of claim 97 , further comprising releasing at least some of the oligonucleotides from the particles.107. The method of claim 97 , further comprising exposing the nucleic acid sequence attached to the oligonucleotides to nucleic acids arising from a plurality of cells which are present in at least some of the plurality of droplets.108. The method of claim 107 , wherein the plurality of cells is present in the plurality of droplets at no more than 1 cell/droplet.109. The method of claim 107 , further comprising lysing at least ...

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Номер записи: 39
31-03-2016 дата публикации

RAPID PRODUCTION OF DROPLETS

Номер: US20160091145A1
Автор: Amstad Esther, Weitz David A.
Принадлежит: President and Fellows of Harvard College

The present invention generally relates to the production of fluidic droplets. Certain aspects of the invention are generally directed to systems and methods for creating droplets by flowing a fluid from a first channel to a second channel through a plurality of side channels. The fluid exiting the side channels into the second channel may form a plurality of droplets, and in some embodiments, at very high droplet production rates. In addition, in some aspects, double or higher-order multiple emulsions may also be formed. In some embodiments, this may be achieved by forming multiple emulsions through a direct, synchronized production method and/or through the formation of a single emulsion that is collected and re-injected into a second microfluidic device to form double emulsions. 1. An apparatus , comprising:a first microfluidic channel;a second microfluidic channel; andat least five side microfluidic channels each connecting the first microfluidic channel with the second microfluidic channel, wherein the first microfluidic channel has a cross-sectional area at least 20 times greater than the smallest cross-sectional area of the at least five side channels.2. An apparatus , comprising:a first, microfluidic channel having a length of at least about 5 mm;a second microfluidic channel substantially parallel to the first microfluidic channel; andat least five side microfluidic channels each connecting the first microfluidic channel with the second microfluidic channel.3. The apparatus of any one of or , comprising at least 10 side microfluidic channels each connecting the first microfluidic channel with the second microfluidic channel.4. The apparatus of any one of - , comprising at least 30 side microfluidic channels each connecting the first microfluidic channel with the second microfluidic channel.5. The apparatus of any one of - , comprising at least 100 side microfluidic channels each connecting the first microfluidic channel with the second microfluidic channel. ...

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Номер записи: 40
21-03-2019 дата публикации

RAPID PRODUCTION OF DROPLETS

Номер: US20190086034A1
Автор: Amstad Esther, Weitz David A.
Принадлежит:

The present invention generally relates to the production of fluidic droplets. Certain aspects of the invention are generally directed to systems and methods for creating droplets by flowing a fluid from a first channel to a second channel through a plurality of side channels. The fluid exiting the side channels into the second channel may form a plurality of droplets, and in some embodiments, at very high droplet production rates. In addition, in some aspects, double or higher-order multiple emulsions may also be formed. In some embodiments, this may be achieved by forming multiple emulsions through a direct, synchronized production method and/or through the formation of a single emulsion that is collected and re-injected into a second microfluidic device to form double emulsions. 147-. (canceled)48. An apparatus , comprising:a first microfluidic channel;a second microfluidic channel;at least five side microfluidic channels each connecting the first microfluidic channel with the second microfluidic channel; anda plurality of auxiliary microfluidic channels connecting to each of the at least five side microfluidic channels.49. The apparatus of claim 48 , comprising at least 10 side channels each connecting the first microfluidic channel with the second microfluidic channel.50. The apparatus of claim 48 , wherein each of the at least five side channels has a length of between about 90% and about 110% of the average length of the side channels.51. The apparatus of claim 48 , wherein the first microfluidic channel and the second microfluidic channel have a distance of separation that is between about 90% and about 110% of the average distance of separation.52. The apparatus of claim 48 , wherein the at least five side channels are positioned such that the distance of separation between any neighboring side channels is between 90% and about 110% of the average distance of separation between neighboring side channels.53. The apparatus of claim 52 , wherein the at least ...

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Номер записи: 41
19-03-2020 дата публикации

MICROPARTICLES, METHODS FOR THEIR PREPARATION AND USE

Номер: US20200085753A1
Автор: Amstad Esther, Mao Angelo S., Mooney David J., Ostafe Raluca, Prodanovich Radivoje, Utech Stefanie, WANG Huanan, Weitz David A., Wilking Connie Chang
Принадлежит: President and Fellows of Harvard College

The invention relates to microparticles comprising a crosslinked gel and methods for making and using same. 142-. (canceled)43. An article , comprising:a tissue scaffold, comprising a plurality of microparticles, the microparticles comprising a core and a shell,a first cell type in the core, anda second cell type in the shell.44. The article of claim 43 , wherein the microparticles comprise a crosslinked gel.45. The article of claim 43 , wherein the microparticles comprise a Ca-crosslinked alginate gel.46. The article of claim 43 , wherein the microparticles have a coefficient of variation in size distribution of from about 0.03 to about 0.05.47. The article of claim 43 , wherein the microparticles comprise an alginate gel or a alginate derivative thereof.48. The article of claim 43 , wherein the first cell type grows in three dimensions within the microparticles.49. The article of claim 43 , wherein the second cell type grows in three dimensions within the microparticles.50. The article of claim 43 , wherein the microparticles further comprise a virus.51. The article of claim 43 , wherein the microparticles further comprise a protein.52. The article of claim 43 , wherein the first cell type at least partially protrudes into the shell.53. The article of claim 43 , wherein the second cell type at least partially protrudes into the core.54. The article of claim 43 , wherein the first cell type at least partially protrudes to an exterior of the microparticles.55. The article of claim 43 , wherein the second cell types at least partially protrudes to an exterior of the microparticles.54. The article of claim 43 , wherein the microparticles further comprise pores.55. The article of claim 43 , wherein the first cell type and the second cell type are present in different concentrations. This application is a divisional of U.S. patent application Ser. No. 15/915,686, filed Mar. 8, 2018, which is a divisional of U.S. patent application Ser. No. 15/035,167, filed May 6, 2016, ...

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Номер записи: 42
12-04-2018 дата публикации

METHODS AND COMPOSITIONS FOR INJECTING HIGH CONCENTRATION AND/OR HIGH VISCOSITY ACTIVE AGENT SOLUTIONS

Номер: US20180098936A1
Автор: DIDIER Jonathan, Ingber Donald E., Weitz David A., ZIERINGER Maximilian
Принадлежит: President and Fellows of Harvard College

Embodiments of various aspects described herein relate to methods and compositions for injecting and/or delivering high viscosity and/or high concentration active agent solutions. In some embodiments, the methods and compositions described herein can be used for subcutaneous administration. 1. A method of producing an injectable composition comprising a high-concentration dose of an active agent , the method comprising: the droplets comprise a first liquid, the first liquid comprising an active agent at a concentration of at least about 50 mg/mL; and', 'the droplets and the injectable carrier liquid are substantially immiscible, thereby producing an injectable composition comprising a high-concentration dose of an active agent., 'forming an emulsion comprising droplets dispersed an injectable carrier liquid, wherein25.-. (canceled)6. A method of producing an injectable composition comprising a high-viscosity dose of an active agent , the method comprising: the droplets comprise a first liquid, the first liquid comprising an active agent and having a viscosity of at least about 20 cP; and', 'the droplets and the injectable carrier liquid are substantially immiscible, thereby producing an injectable composition comprising a high-viscosity dose of an active agent., 'forming an emulsion comprising droplets dispersed in an injectable carrier liquid, wherein751.-. (canceled)52. A vial comprising a composition comprising:droplets comprising a first liquid, the first liquid comprising an active agent at a concentration of at least about 50 mg/mL or higher; anda carrier liquid, wherein the carrier liquid and the droplets are substantially immiscible, and the droplets are dispersed in the carrier liquid.53. A vial comprising a composition comprising:droplets comprising a first liquid, the first liquid comprising an active agent and having a viscosity of at least about 20 cP or higher; anda carrier liquid, wherein the carrier liquid and the droplets are substantially ...

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Номер записи: 43
13-04-2017 дата публикации

COMPARTMENTALISED SCREENING BY MICROFLUIDIC CONTROL

Номер: US20170102381A1
Автор: Ahn Keunho, Bibette Jérôme, Griffiths Andrew David, Link Darren Roy, Weitz David A.
Принадлежит:

The invention describes a method for the identification of compounds which bind to a target component of a biochemical system or modulate the activity of the target, comprising the steps of: a) compartmentalising the compounds into microcapsules together with the target, such that only a subset of the repertoire is represented in multiple copies in any one microcapsule; and b) identifying the compound which binds to or modulates the activity of the target; wherein at least one step is performed under microfluidic control. The invention enables the screening of large repertoires of molecules which can serve as leads for drug development. 1102-. (canceled)103. A method for screening a compound or compounds capable of modulating the activity of a target , comprising the steps of:(a) compartmentalising the compounds into microcapsules, such that only a subset of the repertoire is represented in multiple copies in any one microcapsule;(b) contacting a target having a desired activity with the compound or compounds and monitoring the modulation of an activity of the target by the compound or compounds, wherein one or more of steps a, b and c is performed under microfluidic control; and(c) identifying and sorting the microcapsules which contain the compound(s) having the desired activity using a change in their optical properties.104115-. (canceled)116. The method of claim 103 , wherein the target is compartmentalized together with the compounds in the microcapsules.117. The method of claim 103 , wherein a fluid stream comprising the target is introduced into the microcapsules comprising the compounds.118. The method of claim 103 , further comprising fusing a microcapsule comprising the target with each of the microcapsules comprising the compounds.119. The method of claim 103 , further comprising attaching the repertoire of compounds to microbeads.120. The method of claim 119 , wherein each microbead comprises a detectable tag.121. The method of claim 119 , wherein the ...

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Номер записи: 44
04-04-2019 дата публикации

MICROFLUIDIC DETERMINATION OF IMMUNE AND OTHER CELLS

Номер: US20190101537A1
Автор: Heyman John, Sun Li, Weitz David A., Zhang Xu
Принадлежит: President and Fellows of Harvard College

The present invention generally relates to fluidic droplets and systems and methods for determining immune or other cells. Some aspects of the invention are generally directed to assays that combine sensitive detection of secreted products with detection of target cell death in droplets containing an effector cell, systems and methods to isolate droplets in which one or more cell interactions have occurred, or systems and methods to generate nucleic acid information from cell interactions. In addition, some embodiments of the invention are generally directed to containing two (or more) cells in droplets, e.g., an effector cell and one or more target cells, and determining various interactions between the cells within the droplets, such as whether the effector cell kills the target cell, whether the effector cell releases antibodies, cytokines or other substances that are able to interact with the target cell or are released in the presence of the target cell, or the like. 1. A method , comprising:containing, in a microfluidic droplet, an immune cell, a target cell, a first signaling entity that determines cell viability, and a second signaling entity that determines a cytokine;determining the first signaling entity within the droplet to determine viability of the immune cell and/or the target cell; anddetermining the second signaling entity within the droplet to determine the cytokine within the droplet, wherein the second signaling entity is distinguishable from the first signaling entity.2. The method of claim 1 , wherein the first signaling entity comprises a first fluorescent entity.3. The method of any one of or claim 1 , wherein the first signaling entity comprises calcein and/or a calcein derivative.4. The method of any one of - claim 1 , wherein the first signaling entity comprises an anti-lactose dehydrogenase antibody.5. The method of any one of - claim 1 , wherein the second signaling entity comprises a particle having attached thereto a first antibody ...

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Номер записи: 45
17-07-2014 дата публикации

ASSAYS AND OTHER REACTIONS INVOLVING DROPLETS

Номер: US20140199730A1
Автор: Agresti Jeremy, Chu Liang-yin, Church George, DANTAS Gautam, KIM Jin-Woong, Rowat Amy, SOMMER Morten, Weitz David A.
Принадлежит: President and Fellows of Harvard College

The present invention generally relates to droplets and/or emulsions, such as multiple emulsions. In some cases, the droplets and/or emulsions may be used in assays, and in certain embodiments, the droplet or emulsion may be hardened to form a gel. In some aspects, a heterogeneous assay can be performed using a gel. For example, a droplet may be hardened to form a gel, where the droplet contains a cell, DNA, or other suitable species. The gel may be exposed to a reactant, and the reactant may interact with the gel and/or with the cell, DNA, etc., in some fashion. For example, the reactant may diffuse through the gel, or the hardened particle may liquefy to form a liquid state, allowing the reactant to interact with the cell. As a specific example, DNA contained within a gel particle may be subjected to PCR (polymerase chain reaction) amplification, e.g., by using PCR primers able to bind to the gel as it forms. As the DNA is amplified using PCR, some of the DNA will be bound to the gel via the PCR primer. After the PCR reaction, unbound DNA may be removed from the gel, e.g., via diffusion or washing. Thus, a gel particle having bound DNA may be formed in one embodiment of the invention. 1. A method of whole genome amplification comprising:a. generating a first plurality of droplets comprising polynucleotide primers;b. generating a second plurality of droplets comprising polynucleotides to be amplified, wherein the polynucleotides are obtained from a whole genome;c. merging a first droplet of the first plurality of droplets with a second droplet of the second plurality of droplets to form a coalesced droplet; andd. performing a whole genome amplification reaction on the polynucleotides in the coalesced droplet to form amplicons.2. The method of claim 1 , wherein the amplification reaction comprises amplifying a plurality of different regions of the polynucleotides obtained from the genome.3. The method of claim 1 , wherein the amplification reaction is performed ...

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Номер записи: 46
17-07-2014 дата публикации

ASSAY AND OTHER REACTIONS INVOLVING DROPLETS

Номер: US20140199731A1
Автор: Agresti Jeremy, Chu Liang-yin, Church George, DANTAS Gautam, KIM Jin-Woong, Rowat Amy, SOMMER Morten, Weitz David A.
Принадлежит: President and Fellows of Harvard College

The present invention generally relates to droplets and/or emulsions, such as multiple emulsions. In some cases, the droplets and/or emulsions may be used in assays, and in certain embodiments, the droplet or emulsion may be hardened to form a gel. In some aspects, a heterogeneous assay can be performed using a gel. For example, a droplet may be hardened to form a gel, where the droplet contains a cell, DNA, or other suitable species. The gel may be exposed to a reactant, and the reactant may interact with the gel and/or with the cell, DNA, etc., in some fashion. For example, the reactant may diffuse through the gel, or the hardened particle may liquefy to form a liquid state, allowing the reactant to interact with the cell. As a specific example, DNA contained within a gel particle may be subjected to PCR (polymerase chain reaction) amplification, e.g., by using PCR primers able to bind to the gel as it forms. As the DNA is amplified using PCR, some of the DNA will be bound to the gel via the PCR primer. After the PCR reaction, unbound DNA may be removed from the gel, e.g., via diffusion or washing. Thus, a gel particle having bound DNA may be formed in one embodiment of the invention. 1. A method of polynucleotide amplification comprising:{'sup': '5', 'a. generating a library of droplets comprising a plurality of polynucleotide primers, wherein the library comprises more than 10polynucleotide primers;'}b. generating a set of droplets, wherein the droplets comprise at least one polynucleotide to be amplified;c. merging a first droplet from the library of droplets with a second droplet from the set of droplets to form a coalesced droplet; andd. performing an amplification reaction on at least one polynucleotide in the coalesced droplet to form at least one amplicon.2. The method of claim 1 , wherein the library of droplets comprises more than 10polynucleotide primers.3. The method of claim 1 , wherein the amplification reaction is whole genome amplification.4. The ...

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Номер записи: 47
03-05-2018 дата публикации

ELECTRONIC CONTROL OF FLUIDIC SPECIES

Номер: US20180117585A1
Автор: Ahn Keunho, Cheng Zhengdong, Cristobal-Azkarate Galder, Link Darren Roy, Weitz David A.
Принадлежит:

Various aspects of the present invention relate to the control and manipulation of fluidic species, for example, in microfluidic systems. In one aspect, the invention relates to systems and methods for making droplets of fluid surrounded by a liquid, using, for example, electric fields, mechanical alterations, the addition of an intervening fluid, etc. In some cases, the droplets may each have a substantially uniform number of entities therein. For example, 95% or more of the droplets may each contain the same number of entities of a particular species. In another aspect, the invention relates to systems and methods for dividing a fluidic droplet into two droplets, for example, through charge and/or dipole interactions with an electric field. The invention also relates to systems and methods for fusing droplets according to another aspect of the invention, for example, through charge and/or dipole interactions. In some cases, the fusion of the droplets may initiate or determine a reaction. In a related aspect of the invention, systems and methods for allowing fluid mixing within droplets to occur are also provided. In still another aspect, the invention relates to systems and methods for sorting droplets, e.g., by causing droplets to move to certain regions within a fluidic system. Examples include using electrical interactions (e.g., charges, dipoles, etc.) or mechanical systems (e.g., fluid displacement) to sort the droplets. In some cases, the fluidic droplets can be sorted at relatively high rates, e.g., at about 10 droplets per second or more. Another aspect of the invention provides the ability to determine droplets, or a component thereof, for example, using fluorescence and/or other optical techniques (e.g., microscopy), or electric sensing techniques such as dielectric sensing. 1273-. (canceled)274. A method of sorting droplets in a microfluidic system , the method comprising:providing a series of droplets of fluid surrounded by a liquid and flowing in a ...

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Номер записи: 48
03-05-2018 дата публикации

ASSAYS AND OTHER REACTIONS INVOLVING DROPLETS

Номер: US20180119212A1
Автор: Agresti Jeremy, Chu Liang-yin, Church George, DANTAS Gautam, KIM Jin-Woong, Rowat Amy, SOMMER Morten, Weitz David A.
Принадлежит:

The present invention generally relates to droplets and/or emulsions, such as multiple emulsions. In some cases, the droplets and/or emulsions may be used in assays, and in certain embodiments, the droplet or emulsion may be hardened to form a gel. In some aspects, a heterogeneous assay can be performed using a gel. For example, a droplet may be hardened to form a gel, where the droplet contains a cell, DNA, or other suitable species. The gel may be exposed to a reactant, and the reactant may interact with the gel and/or with the cell, DNA, etc., in some fashion. For example, the reactant may diffuse through the gel, or the hardened particle may liquefy to form a liquid state, allowing the reactant to interact with the cell. As a specific example, DNA contained within a gel particle may be subjected to PCR (polymerase chain reaction) amplification, e.g., by using PCR primers able to bind to the gel as it forms. As the DNA is amplified using PCR, some of the DNA will be bound to the gel via the PCR primer. After the PCR reaction, unbound DNA may be removed from the gel, e.g., via diffusion or washing. Thus, a gel particle having bound DNA may be formed in one embodiment of the invention. 158-. (canceled)59. A composition comprising a plurality of aqueous droplets in a water-in-oil emulsion , wherein the plurality of aqueous droplets comprises a plurality of particles and a plurality of additional particles , wherein a given droplet of the plurality of aqueous droplets comprises (i) a particle from the plurality of particles , wherein the particle comprises one or more reagents for carrying out a nucleic acid amplification reaction on a nucleic acid molecule , and (ii) an additional particle from the plurality of additional particles , wherein the additional particle is (1) a cell comprising the nucleic acid molecule , or (2) a matrix comprising the nucleic acid molecule derived from the cell.60. The composition of claim 59 , wherein some of the plurality of aqueous ...

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Номер записи: 49
25-04-2019 дата публикации

Systems and methods for the collection of droplets and/or other entities

Номер: US20190118182A1
Автор: David A. Weitz, Kiryakos S. Mutafopulos, Thomas HUFNAGEL
Принадлежит: Harvard College

The present invention generally relates to microfluidic devices. In some aspects, various entities, such as droplets or particles, may be contained within a microfluidic device, e.g., within collection chambers or other locations within the device. In some cases, the entities may be released from such locations, e.g., in a sequential pattern, or an arbitrary pattern. In some cases, the entities may be imaged, reacted, analyzed, etc. while contained within the collection chambers. Other aspects are generally directed to methods of making or using such devices, kits involving such devices, or the like.

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Номер записи: 50
12-05-2016 дата публикации

Coalescence of droplets

Номер: US20160129444A1
Автор: David A. Weitz, Donald Aubrecht, Ilke Akartuna, Thomas E. Kodger
Принадлежит: Harvard College

The present invention generally relates to microfiuidics, and, in particular, to systems and methods for coalescing or fusing droplets. In certain aspects, two or more droplets within a microfluidic channel are brought together and caused to coalesce without using electric fields or charges. For example, in certain embodiments, droplets stabilized with a surfactant may be disrupted, e.g., by exposing the droplets to a solvent able to alter the surfactant, which may partially destabilize the droplets and allow them to coalesce. In some instances, the droplets may also be physically disrupted to facilitate coalesce. In addition, in some cases, the positions of one or more droplets may be controlled within a channel using a groove in a wall of the channel. For example, a droplet may at least partially enter the groove such that the position of the droplet is at least partially controlled by the groove.

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Номер записи: 51
11-05-2017 дата публикации

MANIPULATION OF FLUIDS, FLUID COMPONENTS AND REACTIONS IN MICROFLUIDIC SYSTEMS

Номер: US20170128943A1
Автор: Agresti Jeremy, Boukellal Hakim, Fraden Seth, Jia Yanwei, Rowat Amy, Selimovic Seila, Weitz David A.
Принадлежит:

Microfluidic structures and methods for manipulating fluids, fluid components, and reactions are provided. In one aspect, such structures and methods can allow production of droplets of a precise volume, which can be stored/maintained at precise regions of the device. In another aspect, microfluidic structures and methods described herein are designed for containing and positioning components in an arrangement such that the components can be manipulated and then tracked even after manipulation. For example, cells may be constrained in an arrangement in microfluidic structures described herein to facilitate tracking during their growth and/or after they multiply. 179-. (canceled)80. A method of partitioning a slug in a microfluidic network , comprisingproviding a microfluidic network comprising a plurality of sections each comprising a microfluidic inlet, a first region, and a second region, wherein the plurality of sections are in fluid communication with each other;flowing a first slug of volume X through a first section, wherein the first slug comprises a reactive component;partitioning a first subset volume of the first slug at the first region of the first section;flowing the first remaining first slug to the downstream second section;partitioning a second subset volume of the first remaining first slug at the first region of the second section; andsequentially partitioning the second remaining first slug to the downstream sections until the first plug is of zero volume.81. The method of claim 80 , wherein the slug is a droplet.82. The method of claim 81 , wherein the droplet is of a first fluid surrounded by an immiscible fluid.83. The method of claim 81 , wherein the droplet is of a first fluid bound on a solid surface.84. The method of claim 82 , wherein the first fluid is an aqueous solution. The present invention relates generally to microfluidic structures, and more specifically, to microfluidic structures and methods for manipulating fluids, fluid ...

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Номер записи: 52
01-09-2022 дата публикации

FLUID INJECTION

Номер: US20220274072A1
Автор: Abate Adam R., Hung Tony, Mary Pascaline, Weitz David A.
Принадлежит: President and Fellows of Harvard College

The present invention generally relates to systems and methods for the control of fluids and, in some cases, to systems and methods for flowing a fluid into and/or out of other fluids. As examples, fluid may be injected into a droplet contained within a fluidic channel, or a fluid may be injected into a fluidic channel to create a droplet. In some embodiments, electrodes may be used to apply an electric field to one or more fluidic channels, e.g., proximate an intersection of at least two fluidic channels. For instance, a first fluid may be urged into and/or out of a second fluid, facilitated by the electric field. The electric field, in some cases, may disrupt an interface between a first fluid and at least one other fluid. Properties such as the volume, flow rate, etc. of a first fluid being urged into and/or out of a second fluid can be controlled by controlling various properties of the fluid and/or a fluidic droplet, for example curvature of the fluidic droplet, and/or controlling the applied electric field. 135-. (canceled)36. A method , comprising:providing a microfluidic system comprising a first microfluidic channel and a second microfluidic channel contacting the first microfluidic channel at an intersection, wherein the microfluidic system further comprises first and second electrodes positioned to create an electric field within the intersection;providing a first fluidic droplet in the first microfluidic channel and a second fluid in the second microfluidic channel, wherein the first fluidic droplet and the second fluid contact each other forming a fluidic interface at least partially within the intersection andflowing a predetermined volume of the second fluid into the first fluidic droplet by applying a voltage to the first and second electrodes, wherein the volume of the second fluid flowing into the first fluidic droplet is partially controlled by controlling the voltage applied to the first and second electrodes.37. The method of claim 36 , further ...

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Номер записи: 53
02-05-2019 дата публикации

Droplet-Based Method And Apparatus For Composite Single-Cell Nucleic Acid Analysis

Номер: US20190127782A1
Автор: Alexander K. Shalek, Anindita BASU, Aviv Regev, Christopher B. FORD, David A. Weitz, Evan Zane MACOSKO, Hongkun Park, Steven Andrew MCCARROLL
Принадлежит: Broad Institute Inc, Harvard College, Massachusetts Institute of Technology

The present invention generally relates to a combination of molecular barcoding and emulsion-based microfluidics to isolate, lyse, barcode, and prepare nucleic acids from individual cells in a high-throughput manner.

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Номер записи: 54
02-05-2019 дата публикации

Immobilization-based systems and methods for genetic analysis and other applications

Номер: US20190127789A1
Автор: David A. Weitz, Huidan ZHANG, John Heyman
Принадлежит: Harvard College

The present invention generally relates to microfluidics and labeled nucleic acids. Certain aspects are generally directed to containing cells in gels, such as agarose gels, and determining nucleic acids within the cells, e.g., while contained within the gels. The nucleic acids may be, for example, genomic DNA, mRNA, transcriptomes, or the like. In some embodiments, for instance, both genomic DNA and RNA (e.g., as in a transcriptome) from a cell may be determined. In some cases, the nucleic acids may be attached to beads for sequencing or other purposes. Such systems may be useful, for example, for high-throughput sequencing or other applications.

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Номер записи: 55
03-06-2021 дата публикации

Droplet arrays for detection and quantification of analytes

Номер: US20210164971A1
Автор: David A. Weitz, David R. Walt, Limor Cohen, Nai Wen CUI, Yamei Cai
Принадлежит: Brigham and Womens Hospital Inc, Harvard College

The invention provides high-sensitivity methods for detection and quantification of target analytes in liquid samples (e.g., biological or environmental samples). The methods can be multiplexed to allow simultaneous detection and quantification of multiple target analytes that are contained in the same sample. The invention also provides related compositions and kits.

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Номер записи: 56
07-08-2014 дата публикации

Multiple emulsions and techniques for the formation of multiple emulsions

Номер: US20140220350A1
Автор: Alireza ABBASPOURRAD, David A. Weitz, Shin-Hyn Kim
Принадлежит: Harvard College

Multiple emulsions and techniques for the formation of multiple emulsions are generally described. A multiple emulsion, as used herein, describes larger droplets that contain one or more smaller droplets therein. In some embodiments, the larger droplet or droplets may be suspended in a carrying fluid containing the larger droplets that, in turn, contain the smaller droplets. As described below, multiple emulsions can be formed in one step in certain embodiments, with generally precise repeatability, and can be tailored in some embodiments to include a relatively thin layer of fluid separating two other fluids.

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Номер записи: 57
26-05-2016 дата публикации

Systems for Encapsulation of Actives within Droplets and other Compartments

Номер: US20160144329A1
Автор: Abbaspourrad Alireza, Caggioni Marco, Choi Chang-Hyung, Weitz David A, Wesner John Christopher, Zhu Taotao
Принадлежит:

The present invention generally relates to microparticles and, in particular, to systems and methods for encapsulation within microparticles. In one aspect, the present invention is generally directed to microparticles containing entities therein, where the entities contain an agent that can be released from the microparticles, e.g., via diffusion. In some cases, the agent may be released from the microparticles without disruption of the microparticles. The entities may be, for instance, polymeric particles, hydrogel particles, droplets of fluid, etc. The entities may be contained within a fluid that is, in turn, encapsulated within the microparticle. The agent may be released from the entity into the fluid, and then from the fluid through the microparticle. In such fashion, the release of agent from the microparticle may be controlled, e.g., over relatively long time scales. Other embodiments of the present invention are generally directed to methods of making such microparticles, methods of using such microparticles, microfluidic devices for making such microparticles, and the like. 1. An apparatus for producing microparticles , comprising:a first reservoir containing a first liquid, wherein the first liquid is an emulsion comprising a dispersed phase and a continuous phase, the first reservoir being in fluid communication with a plurality of first conduits having exits;a second reservoir containing a second liquid, the second reservoir being in fluid communication with a plurality of second conduits, the second conduits at least partly surrounding the first conduits;a third reservoir containing a third liquid, the third reservoir being in fluid communication with second conduits; anda plurality of third conduits disposed at least in part within the second conduits downstream of the exits of the first conduits, the third conduits positioned to receive the first liquid exiting the first conduits, the second liquid and the third liquid.2. The apparatus of claim 1 , ...

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Номер записи: 58
26-05-2016 дата публикации

Methods for Encapsulation of Actives within Droplets and other Compartments

Номер: US20160144330A1
Автор: Abbaspourrad Alireza, Caggioni Marco, Choi Chang-Hyung, Weitz David A., Wesner John Christopher, Zhu Taotao
Принадлежит:

The present invention generally relates to microparticles and, in particular, to systems and methods for encapsulation within microparticles. In one aspect, the present invention is generally directed to microparticles containing entities therein, where the entities contain an agent that can be released from the microparticles, e.g., via diffusion. In some cases, the agent may be released from the microparticles without disruption of the microparticles. The entities may be, for instance, polymeric particles, hydrogel particles, droplets of fluid, etc. The entities may be contained within a fluid that is, in turn, encapsulated within the microparticle. The agent may be released from the entity into the fluid, and then from the fluid through the microparticle. In such fashion, the release of agent from the microparticle may be controlled, e.g., over relatively long time scales. Other embodiments of the present invention are generally directed to methods of making such microparticles, methods of using such microparticles, microfluidic devices for making such microparticles, and the like. 1. A method for producing microparticles , comprising steps of:providing a first liquid comprising an emulsion, the emulsion comprising a dispersed phase and a continuous phase, to a system of microfluidic devices;introducing a stream of the first liquid into a stream of a second liquid, wherein the continuous phase of the first liquid is substantially immiscible in the second liquid;introducing a stream of a third liquid that surrounds the stream of the second liquid; andforming droplets within the third liquid, wherein each of the droplets comprise a core comprising the first liquid and a shell comprising the second liquid.2. The method of claim 1 , wherein the dispersed phase is substantially miscible with the second liquid.3. The method of claim 1 , further comprising solidifying the shells of the droplets to form microparticles.4. The method of claim 3 , wherein the microparticles ...

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Номер записи: 59
26-05-2016 дата публикации

Compositions Comprising Encapsulated Actives within Droplets and other Compartments

Номер: US20160145535A1
Автор: Abbaspourrad Alireza, Caggioni Marco, Choi Chang-Hyung, Weitz David A., Wesner John Christopher, Zhu Taotao
Принадлежит:

The present invention generally relates to microparticles and, in particular, to systems and methods for encapsulation within microparticles. In one aspect, the present invention is generally directed to microparticles containing entities therein, where the entities contain an agent that can be released from the microparticles, e.g., via diffusion. In some cases, the agent may be released from the microparticles without disruption of the microparticles. The entities may be, for instance, polymeric particles, hydrogel particles, droplets of fluid, etc. The entities may be contained within a fluid that is, in turn, encapsulated within the microparticle. The agent may be released from the entity into the fluid, and then from the fluid through the microparticle. In such fashion, the release of agent from the microparticle may be controlled, e.g., over relatively long time scales. Other embodiments of the present invention are generally directed to methods of making such microparticles, methods of using such microparticles, microfluidic devices for making such microparticles, and the like. 1. A composition comprising a plurality of microparticles , each microcapsule comprising:a single core comprising a liquid emulsion having a continuous phase and a dispersed phase;and a shell surrounding the single core, the shell comprising a wall having a mean wall thickness from about 0.1 μm to about 10 μm.2. The composition of wherein the microcapsules continuous phase comprises a hydrogel.3. The composition according to wherein the dispersed phase retention fraction is greater than about 40%.4. The composition of wherein the dispersed phase comprises a material selected from the group consisting of prop-2-enyl 3-cyclohexylpropanoate claim 1 , (4aR claim 1 ,5R claim 1 ,7aS claim 1 ,9R)-octahydro-2 claim 1 ,2 claim 1 ,5 claim 1 ,8 claim 1 ,8 claim 1 ,9a-hexamethyl-4h-4a claim 1 ,9-methanoazuleno(5 claim 1 ,6-d)-1 claim 1 ,3-dioxole claim 1 , (3aR claim 1 ,5 aS claim 1 ,9aS claim 1 , ...

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Номер записи: 60
17-06-2021 дата публикации

MANIPULATION OF FLUIDS, FLUID COMPONENTS AND REACTIONS IN MICROFLUIDIC SYSTEMS

Номер: US20210178395A1
Автор: Agresti Jeremy, Boukellal Hakim, Fraden Seth, Jia Yanwei, Rowat Amy, Selimovic Seila, Weitz David A.
Принадлежит:

Microfluidic structures and methods for manipulating fluids, fluid components, and reactions are provided. In one aspect, such structures and methods can allow production of droplets of a precise volume, which can be stored/maintained at precise regions of the device. In another aspect, microfluidic structures and methods described herein are designed for containing and positioning components in an arrangement such that the components can be manipulated and then tracked even after manipulation. For example, cells may be constrained in an arrangement in microfluidic structures described herein to facilitate tracking during their growth and/or after they multiply. 120-. (canceled)21. A method for droplet formation , the method comprising:providing a microfluidic network comprising at least a first inlet to a microfluidic channel, wherein the microfluidic channel comprises a first region and a second region;introducing a first fluid comprising first and second reactive components into the microfluidic channel, the first fluid comprising a biological sample and a plurality of beads, each bead comprising a reactive component for binding a target molecule from the biological sample to the bead; andflowing said first fluid into said first and second regions, such that said first fluid encounters an immiscible second fluid in said second region, thereby to form a single droplet of said first fluid surrounded by said second fluid in said second region.22. The method of claim 21 , wherein each of the plurality of beads comprises a reactive component for binding to a target molecule from a biological sample.23. The method of claim 22 , wherein the one or more droplets comprises a biological sample.24. The method of claim 23 , wherein the target molecule comprises DNA or RNA.25. The method of claim 22 , wherein the target molecule is a single biological molecule.26. The method of claim 22 , wherein the target molecule is a protein.27. The method of claim 21 , wherein the second ...

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Номер записи: 61
01-06-2017 дата публикации

SCALE-UP OF MICROFLUIDIC DEVICES

Номер: US20170151536A1
Автор: Abate Adam R., Romanowsky Mark, Weitz David A.
Принадлежит:

Parallel uses of microfluidic methods and devices for focusing and/or forming discontinuous sections of similar or dissimilar size in a fluid are described. In some aspects, the present invention relates generally to flow-focusing-type technology, and also to microfluidics, and more particularly parallel use of microfluidic systems arranged to control a dispersed phase within a dispersant, and the size, and size distribution, of a dispersed phase in a multi-phase fluid system, and systems for delivery of fluid components to multiple such devices. 178-. (canceled)79. A system for forming droplets in microfluidic channels in parallel , comprising a distribution channel connecting a source of a dispersing fluid to a plurality of dispersing fluid outlets , wherein a given dispersing fluid outlet of the plurality is directly fluidly connected to a given microfluidic interconnected region of a plurality of microfluidic interconnected regions arranged in parallel , wherein a given microfluidic interconnected region of the plurality of microfluidic interconnected regions joins a subject fluid channel for carrying a subject fluid and a dispersing fluid channel for carrying the dispersing fluid , wherein at least a portion of an outer wall of the given microfluidic interconnected region and a portion of an outer wall of the subject fluid channel are portions of a single integral unit.80. The system of claim 79 , wherein the outer wall of the interconnected region is exterior of the outer wall of the subject fluid channel.81. The system of claim 79 , further comprising a dimensionally-restricted section formed by extensions extending from the outer wall of the given microfluidic interconnected region into the given microfluidic interconnected region claim 79 , wherein the dimensionally-restricted section is positioned adjacent to a subject fluid channel outlet.82. The system of claim 81 , wherein the dimensionally-restricted section has a shape which causes the dispersing ...

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Номер записи: 62
11-06-2015 дата публикации

ULTRASENSITIVE DETECTION AND CHARACTERIZATION OF CLUSTERED KRAS MUTATIONS USING PEPTIDE NUCLEIC ACID CLAMP PCR IN DROP-BASED MICROFLUIDICS

Номер: US20150159224A1
Автор: Davey Neil, SPERLING Ralph, Weitz David A., Zhang Huidan
Принадлежит:

This disclosure employs the combination of a microfluidics platform and drop-based digital polymerase chain reaction (dPCR) to create a breakthrough technology that enables the detection of CTC genes and the isolation of single CTCs from the blood. In the first method, cDNA molecules from lysed CTCs are amplified in microfluidic drops and detected via fluorescence signal. In the second method, intact single CTCs are encapsulated, and amplification-positive drops are sorted from the remaining cells. To demonstrate the clinical utility of our technology, mutations in the KRAS gene in colorectal cancer are analyzed to study resistance to EGFR-based treatment as a test case. The methods herein present robust techniques for both the diagnosis and treatment of cancers, as well as for the obtainment of a pure CTC sample from billions of other cells in the blood. 1. A method for diagnosing cancer in a person or animal , comprising:Obtaining or preparing a sample comprising cDNAs of a plurality of genes of the person or animal;encapsulating the cDNAs into discrete droplets, wherein statistically each of the discrete droplets contains at most one of the cDNAs;amplifying the cDNAs in the droplets; anddetermining whether the droplets contain a cDNA of a mutation of a V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) gene.2. The method of claim 1 , further comprising sorting the droplets.3. The method of claim 1 , wherein the sample is a whole blood sample.4. The method of claim 1 , wherein obtaining the sample comprises reverse transcribing mRNAs.5. The method of claim 1 , wherein the cancer is colorectal cancer.6. The method of claim 1 , wherein the cancer is prostate cancer.7. The method of claim 1 , wherein the mutation is codon 12 or codon 13 of the KRAS gene.8. The method of claim 1 , wherein the mutation is alteration of a guanine in the KRAS gene.9. The method of claim 1 , wherein determining whether the droplets contain a cDNA of a mutation of the KRAS gene is ...

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Номер записи: 63
07-06-2018 дата публикации

SYSTEMS AND METHODS FOR EPIGENETIC SEQUENCING

Номер: US20180155777A1
Автор: BERNSTEIN BRADLEY E., Ram Oren, Rotem Assaf, Weitz David A.
Принадлежит:

The present invention generally relates to microfluidics and/or epigenetic sequencing. In one set of embodiments, cells contained within a plurality of microfluidic droplets are lysed and the DNA (e.g., from nucleosomes) within the droplets are labeled, e.g., with adapters containing an identification sequence. The adapters may also contain other sequences, e.g., restriction sites, primer sites, etc., to assist with later analysis. After labeling with adapters, the DNA from the different cells may be combined and analyzed, e.g., to determine epigenetic information about the cells. For example, the DNA may be separated on the basis of certain modifications (e.g., methylation), and the DNA from the separated nucleosomes may be sequenced using techniques such as chromatin immunoprecipitation (“ChIP”). In some cases, the DNA sequences may also be aligned with genomes, e.g., to determine which portions of the genome were epigenetically modified, e.g., via methylation. 1111-. (canceled)112. A method for providing a composition comprising a plurality of droplets comprising nucleic acid sequence adapters comprising an identification sequence and a restriction site , comprising:providing the adapters on a solid support binding the adapters to nucleic acids;producing droplets in a microfluidic device, the device comprising at least two microfluidic channels combined via a junction, wherein the fluid forming the droplets is substantially immiscible with the carrier fluid surrounding the droplets; andencapsulating the adapters, the nucleic acids and the solid support in at least some of the droplets.113. The method of claim 112 , wherein the junction is selected from the group consisting of a T-junction claim 112 , a Y-junction claim 112 , a channel-within-a-channel junction claim 112 , a cross junction claim 112 , and a flow-focus junction.114. The method of claim 112 , wherein the adapters comprise primer sites.115. The method of claim 114 , wherein the primer sites comprise ...

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Номер записи: 64
07-06-2018 дата публикации

SYSTEMS AND METHODS FOR EPIGENETIC SEQUENCING

Номер: US20180155778A1
Автор: BERNSTEIN BRADLEY E., Ram Oren, Rotem Assaf, Weitz David A.
Принадлежит:

The present invention generally relates to microfluidics and/or epigenetic sequencing. In one set of embodiments, cells contained within a plurality of microfluidic droplets are lysed and the DNA (e.g., from nucleosomes) within the droplets are labeled, e.g., with adapters containing an identification sequence. The adapters may also contain other sequences, e.g., restriction sites, primer sites, etc., to assist with later analysis. After labeling with adapters, the DNA from the different cells may be combined and analyzed, e.g., to determine epigenetic information about the cells. For example, the DNA may be separated on the basis of certain modifications (e.g., methylation), and the DNA from the separated nucleosomes may be sequenced using techniques such as chromatin immunoprecipitation (“ChIP”). In some cases, the DNA sequences may also be aligned with genomes, e.g., to determine which portions of the genome were epigenetically modified, e.g., via methylation. 1111-. (canceled)112. A method comprising:providing a solution comprising a plurality of nucleic acid sequences originating from a plurality of cells, at least some of the nucleic acid sequences being attached to an adapter, the adapter comprising an identification sequence, wherein sequences originating from the same cell contain identical identification sequences, and sequences originating from different cells contain different identification sequences, the adapter further comprising a primer site and a cleavage site, which is cleavable by a restriction endonuclease, wherein the adapters are present on a solid support; andamplifying at least some of the sequences.113. The method of claim 112 , comprising cleaving the adapter at the cleavage site.114. The method of claim 112 , wherein the solution is contained within a droplet.115. The method of claim 112 , wherein the plurality of nucleic acid sequences comprises DNA.116. The method of claim 112 , wherein the plurality of nucleic acid sequences comprises ...

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Номер записи: 65
23-05-2019 дата публикации

DETERMINATION OF RNA IN BLOOD OR OTHER FLUIDS

Номер: US20190153427A1
Автор: Cui Nai Wen, Weitz David A., Zhang Huidan
Принадлежит: President and Fellows of Harvard College

The present invention generally relates to systems and methods for determining RNA in blood or other fluids. In certain embodiments, blood or other fluids may be treated to isolate or separate RNA, for example, from DNA, cells, and other material. In some cases, the RNA may arise from bacteria or other pathogens or foreign organisms that may be found within the blood or other fluid. In some cases, RNA stabilizing reagents, such as ammonium sulfate, may be added to stabilize RNA, then cells within the blood may be lysed to release the RNA (and other materials) from the cells, thereby producing a lysate. The lysate may be treated, e.g., to separate nucleic acids from other components within the lysate, and in some cases, DNA may be degraded, e.g., using DNAses or other suitable enzymes, leaving behind the RNA. The RNA can then be studied, purified, analyzed, amplified, stored, or the like. 1. A method , comprising:adding an RNA-stabilizing reagent to a blood sample;lysing cells within the blood sample to produce a lysate;degrading DNA within the lysate; andseparating RNA from the lysate.2. The method of claim 1 , wherein the RNA-stabilizing reagent comprises ammonium sulfate.3. The method of any one of or claim 1 , wherein the ammonium sulfate is added to produce a final concentration in the blood sample of no more than 20 g/100 mL.4. The method of any one of - claim 1 , wherein the ammonium sulfate is added to produce a final concentration in the blood sample of no more than 64 mM.5. The method of any one of - claim 1 , wherein lysing cells within the blood sample comprises exposing the cells within the blood sample to proteinase K.6. The method of any one of - claim 1 , wherein lysing cells within the blood sample comprises exposing the cells within the blood sample to lysozyme.7. The method of any one of - claim 1 , wherein degrading DNA within the lysate comprises exposing the lysate to a nonspecific endonuclease.8. The method of any one of - claim 1 , wherein ...

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Номер записи: 66
04-09-2014 дата публикации

Manipulation of fluids, fluid components and reactions in microfluidic systems

Номер: US20140246098A1
Автор: Amy Rowat, David A. Weitz, Hakim Boukellal, Jeremy Agresti, Seila Selimovic, Seth Fraden, Yanwei Jia
Принадлежит: BRANDEIS UNIVERSITY, Harvard College

Microfluidic structures and methods for manipulating fluids, fluid components, and reactions are provided. In one aspect, such structures and methods can allow production of droplets of a precise volume, which can be stored/maintained at precise regions of the device. In another aspect, microfluidic structures and methods described herein are designed for containing and positioning components in an arrangement such that the components can be manipulated and then tracked even after manipulation. For example, cells may be constrained in an arrangement in microfluidic structures described herein to facilitate tracking during their growth and/or after they multiply.

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Номер записи: 67
16-06-2016 дата публикации

STRUCTURALLY COLORED MATERIALS WITH SPECTRALLY SELECTIVE ABSORBING COMPONENTS AND METHODS FOR MAKING THE SAME

Номер: US20160168386A1
Автор: Aizenberg Joanna, Burgess Ian, KOAY Natalie, KOLLE Mathias, PHILLIPS Katherine, SHIRMAN Tanya, Utech Stefanie, VOGEL Nicolas, Weitz David A.
Принадлежит: President and Fellows of Harvard College

A structurally colored pigment is described that contains a plurality of photonic crystal particles dispersed in a medium, where each photonic crystal particles contains a plurality of spectrally selective absorbing components dispersed within the photonic crystal particle. In certain embodiments, each photonic crystal particle has a predetermined minimum number of repeat units of the photonic crystal structure. The structurally colored material provides improved reflectance, long-term stability, and control of the desired optical effects. The fabrication techniques described herein also provide high throughput and high yield allowing use in wide ranging applications from cosmetics, paints, signs, sensors, to packaging material. 1. A pigment comprising:a plurality of photonic crystal particles dispersed in a medium, each photonic crystal particles containing a plurality of spectrally selective absorbing components dispersed within the photonic crystal particle that selectively absorb electromagnetic radiation without substantially absorbing electromagnetic radiation near the resonant wavelength of the photonic crystal particle, whereineach photonic crystal particle has a predetermined minimum number of repeat units of the photonic crystal structure, wherein the predetermined minimum number of repeat units is related to the resonant wavelength, the full-width at half maximum of the resonant wavelength, and the refractive index contrast in the photonic crystal.2. The pigment of claim 1 , wherein the photonic crystal structure is an inverse or a direct opal structure.3. The pigment of claim 1 , wherein the spectrally selective absorbing components are gold nanoparticles or silver nanoparticles.4. The pigment of claim 1 , wherein the collection exhibits a red structural color.5. The pigment of claim 1 , wherein the photonic crystal particles are bricks or spheres.6. The pigment of claim 1 , wherein the photonic crystal structure is an inverse opal structure and the ...

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Номер записи: 68
30-05-2019 дата публикации

FORMATION OF COLLOIDS OR GELS WITHIN DROPLETS

Номер: US20190160445A1
Автор: Cui Nai Wen, Lei Fengyang, llios Eleftherios Paschalis, Weitz David A., Zhang Huidan
Принадлежит:

The present invention generally relates to microfluidic droplets and, including forming gels within microfluidic droplets. In some aspects, a fluid containing agarose or other gel precursors is transported into a microfluidic droplet, and caused to harden within the droplet, e.g., to form a gel particle contained within the microfluidic droplet. Surprisingly, a discrete gel particle may be formed even if the fluid containing the agarose or other gel precursor, and the fluid contained within the microfluidic droplet, are substantially immiscible. Other aspects of the present invention are generally directed to techniques for making or using such gels within microfluidic droplets, kits containing such gels within microfluidic droplets, or the like. 1. A method , comprising:transporting a first fluid containing agarose at at least 0.5 vol % into a microfluidic droplet containing a second fluid; andcausing the agarose gel to solidify to form a gel particle contained within the second fluid in the microfluidic droplet.2. The method of claim 1 , wherein the first fluid contains agarose at at least 0.5 vol %.3. The method of any one of or claim 1 , wherein the first fluid and the second fluid are substantially miscible.4. The method of any one of - claim 1 , wherein the microfluidic droplet containing the second fluid is contained in a carrying fluid.5. The method of claim 4 , wherein the carrying fluid completely surrounds the microfluidic droplet.6. The method of any one of - claim 4 , wherein the microfluidic droplet has an average diameter of less than about 1 micrometer.7. The method of any one of - claim 4 , wherein transporting the first fluid into the microfluidic droplet comprises applying an electric field to an interface between the first fluid and the microfluidic droplet.8. The method of claim 7 , wherein the electric field is applied using two electrodes positioned on the same side of the second microfluidic channel.9. The method of claim 7 , wherein the ...

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Номер записи: 69
21-05-2020 дата публикации

ASSAYS AND OTHER REACTIONS INVOLVING DROPLETS

Номер: US20200157593A1
Автор: Agresti Jeremy, Chu Liang-yin, Church George M., DANTAS Gautam, KIM Jin-Woong, Rowat Amy, SOMMER Morten, Weitz David A.
Принадлежит: President and Fellows of Harvard College

The present invention generally relates to droplets and/or emulsions, such as multiple emulsions. In some cases, the droplets and/or emulsions may be used in assays, and in certain embodiments, the droplet or emulsion may be hardened to form a gel. In some aspects, a heterogeneous assay can be performed using a gel. For example, a droplet may be hardened to form a gel, where the droplet contains a cell, DNA, or other suitable species. The gel may be exposed to a reactant, and the reactant may interact with the gel and/or with the cell, DNA, etc., in some fashion. For example, the reactant may diffuse through the gel, or the hardened particle may liquefy to form a liquid state, allowing the reactant to interact with the cell. As a specific example, DNA contained within a gel particle may be subjected to PCR (polymerase chain reaction) amplification, e.g., by using PCR primers able to bind to the gel as it forms. As the DNA is amplified using PCR, some of the DNA will be bound to the gel via the PCR primer. After the PCR reaction, unbound DNA may be removed from the gel, e.g., via diffusion or washing. Thus, a gel particle having bound DNA may be formed in one embodiment of the invention. 120-. (canceled)21. A composition , comprising:a collection of particles, wherein said collection of particles comprises a library of distinguishable nucleic acid entities, wherein a particle of said collection of particles comprises a nucleic acid entity of said library of distinguishable nucleic acid entities, and wherein said particle is surrounded by an aqueous fluid.22. The composition of claim 21 , wherein said aqueous fluid comprises a nucleic acid primer.23. The composition of claim 21 , wherein said particle is configured to liquefy.24. The composition of claim 23 , wherein said particle is configured to liquefy upon application of a stimulus.25. The composition of claim 24 , wherein said stimulus comprises one or more of a change in temperature claim 24 , pH claim 24 , or ...

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Номер записи: 70
01-07-2021 дата публикации

FLUID INJECTION

Номер: US20210197146A1
Автор: Abate Adam R., Hung Tony, Mary Pascaline, Weitz David A.
Принадлежит: President and Fellows of Harvard College

The present invention generally relates to systems and methods for the control of fluids and, in some cases, to systems and methods for flowing a fluid into and/or out of other fluids. As examples, fluid may be injected into a droplet contained within a fluidic channel, or a fluid may be injected into a fluidic channel to create a droplet. In some embodiments, electrodes may be used to apply an electric field to one or more fluidic channels, e.g., proximate an intersection of at least two fluidic channels. For instance, a first fluid may be urged into and/or out of a second fluid, facilitated by the electric field. The electric field, in some cases, may disrupt an interface between a first fluid and at least one other fluid. Properties such as the volume, flow rate, etc. of a first fluid being urged into and/or out of a second fluid can be controlled by controlling various properties of the fluid and/or a fluidic droplet, for example curvature of the fluidic droplet, and/or controlling the applied electric field. 127-. (canceled)28. A method , comprising:providing a microfluidic system comprising a first microfluidic channel and a second microfluidic channel contacting the first microfluidic channel at an intersection;providing a first fluid in the first microfluidic channel and a second fluid in the second microfluidic channel, wherein the first fluid and the second fluid contact each other at least partially within the intersection to define a fluidic interface; andurging the second fluid to enter the first microfluidic channel, wherein, when an electric field is applied to the interface, the second fluid is at least partially prevented from entering the first microfluidic channel.29. The method of claim 28 , wherein the first fluid is present within the first microfluidic channel as a plurality of droplets contained within a carrying fluid.30. The method of claim 28 , wherein the second droplet is urged to form droplets within the first microfluidic channel in ...

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Номер записи: 71
21-06-2018 дата публикации

Assays and other reactions involving droplets

Номер: US20180171373A1
Автор: Amy Rowat, David A. Weitz, Gautam Dantas, George Church, Jeremy Agresti, Jin-woong Kim, Liang-Yin Chu, Morten Sommer
Принадлежит: Harvard College

The present invention generally relates to droplets and/or emulsions, such as multiple emulsions. In some cases, the droplets and/or emulsions may be used in assays, and in certain embodiments, the droplet or emulsion may be hardened to form a gel. In some aspects, a heterogeneous assay can be performed using a gel. For example, a droplet may be hardened to form a gel, where the droplet contains a cell, DNA, or other suitable species. The gel may be exposed to a reactant, and the reactant may interact with the gel and/or with the cell, DNA, etc., in some fashion. For example, the reactant may diffuse through the gel, or the hardened particle may liquefy to form a liquid state, allowing the reactant to interact with the cell. As a specific example, DNA contained within a gel particle may be subjected to PCR (polymerase chain reaction) amplification, e.g., by using PCR primers able to bind to the gel as it forms. As the DNA is amplified using PCR, some of the DNA will be bound to the gel via the PCR primer. After the PCR reaction, unbound DNA may be removed from the gel, e.g., via diffusion or washing. Thus, a gel particle having bound DNA may be formed in one embodiment of the invention.

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Номер записи: 72
30-06-2016 дата публикации

IN VITRO EVOLUTION IN MICROFLUIDIC SYSTEMS

Номер: US20160186256A1
Автор: Ahn Keunho, Bibette Jérôme, Griffiths Andrew David, Link Darren R., Weitz David A.
Принадлежит:

The invention describes a method for isolating one or more genetic elements encoding a gene product having a desired activity, comprising the steps of: (a) compartmentalising genetic elements into microcapsules; and (b) sorting the genetic elements which express the gene product having the desired activity; wherein at least one step is under microfluidic control. The invention enables the in vitro evolution of nucleic acids and proteins by repeated mutagenesis and iterative applications of the method of the invention. 1107.-. (canceled)108. A method for detecting a selectable change within a microcapsule , comprising the steps of:forming a plurality of aqueous microcapsules in a fluorinated oil, wherein each aqueous microcapsule comprises a cell and a first genetic element;incubating the microcapsules to cause the first genetic element to interact with a molecule associated with the cell; anddetecting a selectable change in one or more of the microcapsules.109. The method of claim 108 , wherein the fluorinated oil comprises a fluorinated polymer surfactant.110. The method of claim 109 , further comprising pooling the microcapsules into one or more common compartments such that a portion of the plurality of microcapsules contact each other but do not fuse.111. The method of claim 108 , wherein the selectable change comprises an optical signal.112. The method of claim 108 , wherein the first genetic element comprises one or more primer sequences for amplification.113. The method of claim 108 , wherein the cell is an immune cell.114. The method of claim 113 , wherein the immune cell is a B-cell.115. The method of claim 108 , wherein the molecule is a reporter protein.116. The method of claim 108 , wherein the molecule comprises a gene.117. The method of claim 116 , further comprising expressing the gene to form a gene product.118. The method of claim 117 , wherein the gene product is a protein.119. The method of claim 119 , wherein the protein is an antibody.120. The ...

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Номер записи: 73
29-06-2017 дата публикации

SYSTEMS AND METHODS FOR BIOMIMETIC FLUID PROCESSING

Номер: US20170183616A1
Автор: Italiano Joseph E., Mazutis Linas, Thon Jonathan N., Weitz David A.
Принадлежит:

Systems and methods generating physiologic models that can produce functional biological substances are provided. In some aspects, a system includes a substrate and a first and second channel formed therein. The channels extend longitudinally and are substantially parallel to each other. A series of apertures extend between the first channel and second channel to create a fluid communication path passing through columns separating the channels that extends further along the longitudinal dimension than other dimensions. The system also includes a first source configured to selectively introduce into the first channel a first biological composition at a first channel flow rate and a second source configured to selectively introduce into the second channel a second biological composition at a second channel flow rate, wherein the first channel flow rate and the second channel flow rate create a differential configured to generate physiological shear rates within a predetermined range in the channels. 1. A biomimetic microfluidic system comprising:a substrate;a first channel formed in the substrate, the first channel extending from a first input to a first output along a longitudinal dimension and extending along a first transverse dimension;a second channel formed in the substrate, the second channel extending from a second input to a second output along the longitudinal dimension and extending along the first transverse dimension, wherein the first and second channels extend substantially parallel along the longitudinal dimension and are separated by columns extending along a second transverse dimension;a series of apertures formed in the columns separating the first channel and second channel, wherein each of the series of apertures extend along the longitudinal dimension further than in the first transverse direction and the second transverse direction and are positioned proximal to a first portion of the substrate and extend from the first channel to the second ...

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Номер записи: 74
29-06-2017 дата публикации

ASSAYS AND OTHER REACTIONS INVOLVING DROPLETS

Номер: US20170183701A1
Автор: Agresti Jeremy, Chu Liang-yin, Church George, DANTAS Gautam, KIM Jin-Woong, Rowat Amy, SOMMER Morten, Weitz David A.
Принадлежит:

The present invention generally relates to droplets and/or emulsions, such as multiple emulsions. In some cases, the droplets and/or emulsions may be used in assays, and in certain embodiments, the droplet or emulsion may be hardened to form a gel. In some aspects, a heterogeneous assay can be performed using a gel. For example, a droplet may be hardened to form a gel, where the droplet contains a cell, DNA, or other suitable species. The gel may be exposed to a reactant, and the reactant may interact with the gel and/or with the cell, DNA, etc., in some fashion. For example, the reactant may diffuse through the gel, or the hardened particle may liquefy to form a liquid state, allowing the reactant to interact with the cell. As a specific example, DNA contained within a gel particle may be subjected to PCR (polymerase chain reaction) amplification, e.g., by using PCR primers able to bind to the gel as it forms. As the DNA is amplified using PCR, some of the DNA will be bound to the gel via the PCR primer. After the PCR reaction, unbound DNA may be removed from the gel, e.g., via diffusion or washing. Thus, a gel particle having bound DNA may be formed in one embodiment of the invention. 1. A method for nucleic acid processing , comprising:a. directing a first phase comprising amplification reagents along a first channel and a second phase comprising an oil along a second channel to an intersection of said first channel and said second channel, wherein said first phase is immiscible with said second phase;b. at said intersection, generating a plurality of droplets comprising individual droplets each having (i) a single cell comprising a first polynucleotide comprising a nucleic acid sequence, and (ii) said amplification reagents; andc. using said amplification reagents, subjecting said first polynucleotide to nucleic acid amplification to generate a second polynucleotide comprising a sequence derived from said nucleic acid sequence.2. The method of claim 1 , wherein ...

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Номер записи: 75
29-06-2017 дата публикации

PARTICLE-ASSISTED NUCLEIC ACID SEQUENCING

Номер: US20170183715A1
Автор: Abate Adam R., Weitz David A.
Принадлежит:

This invention generally relates to particle-assisted nucleic acid sequencing. In some embodiments, sequencing may be performed in a microfluidic device, which can offer desirable properties, for example, minimal use of reagents, facile scale-up, and/or high throughput. In one embodiment, a target nucleic acid may be exposed to particles having nucleic acid probes. By determining the binding of the particles to the target nucleic acid, the sequence of the target nucleic acid (or at least a portion of the target nucleic acid) can be determined. The target nucleic acid may be encapsulated within a fluidic droplet with the particles having nucleic acid probes, in certain instances. In some cases, the sequence of the target nucleic acid may be determined, based on binding of the particles, using sequencing by hybridization (SBH) algorithms or other known techniques. 183-. (canceled)84. A method of sequencing a target nucleic acid , the method comprising:passing a microfluidic droplet containing a target nucleic acid through a microfluidic channel;exposing the target nucleic acid to a plurality of particles, at least some of which have nucleic acid probes fastened thereto, wherein at least some of the particles are contained within said microfluidic droplet containing the target nucleic acid and said nucleic acid probes comprises a nucleic acid probe that can hybridize with said target nucleic acid, and wherein the particles comprise a gel;hybridizing said nucleic acid probes to the target nucleic acid; andat least partially determining the sequence of the target nucleic acid.85. The method of claim 84 , wherein at least one of the particles is associated with at least one identification entity.86. The method of claim 85 , wherein said at least partially determining the sequence of the target nucleic acid comprises determining the at least one identification entity.87. The method of claim 85 , further comprising determining the at least one identification entity.88. The ...

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Номер записи: 76
09-07-2015 дата публикации

CONTROL OF ENTITIES SUCH AS DROPLETS AND CELLS USING ACOUSTIC WAVES

Номер: US20150192546A1
Автор: Franke Thomas, Weitz David A.
Принадлежит:

The present invention generally relates to manipulation of entities using acoustic waves. For example, by applying acoustic waves to a surface containing entities such as particles, cells, droplets, etc., the entities may be manipulated in various ways on the surface. The surface acoustic waves may be created using a surface acoustic wave generator such as an interdigitated transducer, and/or a material such as a piezoelectric substrate. In some cases, two or more acoustic waves may be applied, and the waves may interfere to create standing waves. The standing waves can be manipulated to manipulate the entities on the surface. For instance, the frequencies of the surface acoustic waves may be slightly mismatched to cause travelling standing waves to occur, which may be used to align the entities. 1. A method , comprising:providing a plurality of entities in a fluid flowing at an average fluid velocity; andaligning at least some of the entities by applying a first acoustic wave and a second acoustic wave to at least a portion of the fluid, wherein the first acoustic wave and the second acoustic wave interfere to create a standing acoustic wave having a nodal propagation velocity within about 20% of the average fluid velocity.2. The method of claim 1 , wherein at least some of the plurality of entities are droplets.3. The method of claim 2 , wherein the droplets are substantially monodisperse.4. The method of any one of or claim 2 , wherein at least some of the droplets are substantially immiscible in the fluid.54. The method of any one of - claims 1 , wherein at least some of the plurality of entities are particles.65. The method of any one of - claims 1 , wherein at least some of the plurality of entities are cells.76. The method of any one of - claims 1 , wherein at least some of the plurality of entities comprise cells.87. The method of any one of - claims 1 , wherein the first acoustic wave has an average frequency of between about 130 MHz and about 160 MHz.98. ...

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Номер записи: 77
07-07-2016 дата публикации

CONTROL OF EMULSIONS, INCLUDING MULTIPLE EMULSIONS

Номер: US20160193574A1
Автор: Abate Adam R., HOLTZE Christian, Rotem Assaf, Weitz David A.
Принадлежит:

The present invention generally relates to emulsions, and more particularly, to double and other multiple emulsions. Certain aspects of the present invention are generally directed to the creation of double emulsions and other multiple emulsions at a common junction of microfluidic channels. In some cases, the microfluidic channels at the common junction may have substantially the same hydrophobicity. In one set of embodiments, a device may include a common junction of six or more channels, where a first fluid flows through one channel, a second fluid flows through two channels, and a third or carrying fluid flows through two more channels, such that a double emulsion of a first droplet of the first fluid, contained in a second droplet of the second fluid, contained by the carrying fluid, flows away from the common junction through a sixth channel. Other aspects of the invention are generally directed to methods of making and using such systems, kits involving such systems, emulsions created using such systems, or the like. 145-. (canceled)46. A method of creating an emulsion encapsulating a species , the method comprising:providing a microfluidic device comprising a first junction of microfluidic channels comprising at least a first, second, and third microfluidic channels in fluidic communication, the first junction in fluid communication at an interface with a second junction of microfluidic channels comprising at least fourth, fifth, and sixth microfluidic channels in fluid communication, each of the first, second, and third microfluidic channels having a respective cross-sectional area at the first junction and each of the fourth, fifth, and sixth microfluidic channels having a respective cross-sectional area at a second junction, wherein the interface has a cross-sectional area smaller than the smallest cross-sectional areas of the fourth, fifth, and sixth microfluidic channels; and creating an emulsion encapsulating a species at the first and second junctions ...

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Номер записи: 78
20-06-2019 дата публикации

Formation and control of fluidic species

Номер: US20190184398A1
Автор: Darren Roy Link, David A. Weitz, Manuel Marquez-Sanchez, Zhengdong Cheng
Принадлежит: Harvard College

This invention generally relates to systems and methods for the formation and/or control of fluidic species, and articles produced by such systems and methods. In some cases, the invention involves unique fluid channels, systems, controls, and/or restrictions, and combinations thereof. In certain embodiments, the invention allows fluidic streams (which can be continuous or discontinuous, i.e., droplets) to be formed and/or combined, at a variety of scales, including microfluidic scales. In one set of embodiments, a fluidic stream may be produced from a channel, where a cross-sectional dimension of the fluidic stream is smaller than that of the channel, for example, through the use of structural elements, other fluids, and/or applied external fields, etc. In some cases, a Taylor cone may be produced. In another set of embodiments, a fluidic stream may be manipulated in some fashion, for example, to create tubes (which may be hollow or solid), droplets, nested tubes or droplets, arrays of tubes or droplets, meshes of tubes, etc. In some cases, droplets produced using certain embodiments of the invention may be charged or substantially charged, which may allow their further manipulation, for instance, using applied external fields. Non-limiting examples of such manipulations include producing charged droplets, coalescing droplets (especially at the microscale), synchronizing droplet formation, aligning molecules within the droplet, etc. In some cases, the droplets and/or the fluidic streams may include colloids, cells, therapeutic agents, and the like.

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Номер записи: 79
20-06-2019 дата публикации

MICROFLUIDIC SEQUENCING TECHNIQUES

Номер: US20190185800A1
Автор: Cui Nai Wen, Weitz David A., Zhang Huidan
Принадлежит: President and Fellows of Harvard College

The present invention generally relates to microfluidics and, in some embodiments, to the determination of cells. In some aspects, primers able to introduce restriction sites into certain amplified nucleic acids are used. For example, the primers may introduce restriction sites into normal (wild-type) nucleic acids, but be unable to introduce restriction sites into mutant nucleic acids, e.g., due to a mismatch in the nucleic acid sequences caused by the mutant. After amplification, the nucleic acids may be exposed to a suitable restriction enzyme, which may cleave normal nucleic acids but not the mutant nucleic acids. In this way, mutant nucleic acids may be relatively quickly identified. In some embodiments, cells may be contained within microfluidic droplets and assayed to determine the mutant cells. In certain cases, for example, the nucleic acids may be amplified within droplets and attached to suitable tags, e.g., prior to breaking or merging the droplets and sequencing of the nucleic acids. 1. A method , comprising:lysing cells contained within microfluidic droplets to release nucleic acids;amplifying the released nucleic acids within the droplets using primers that introduce restriction sites during amplification to produce amplicons;bonding nucleic acid tags to at least some of the amplicons within the droplets;releasing the amplicons from the droplets;exposing the amplicons to a restriction enzyme; andsequencing the amplicons.2. The method of claim 1 , wherein the cells comprise mammalian cells.3. The method of any one of or claim 1 , wherein the cells comprise human cells.4. The method of any one of - claim 1 , wherein the released nucleic acids comprise genomic DNA.5. The method of any one of - claim 1 , wherein the restriction site introduced by the primer is cleavable by the restriction enzyme.6. The method of any one of - claim 1 , wherein the restriction enzyme is EcoRI.7. The method of any one of - claim 1 , wherein the restriction enzyme is AlwNI.8. ...

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Номер записи: 80
25-09-2014 дата публикации

SYSTEMS AND METHODS FOR SPRAY DRYING IN MICROFLUIDIC AND OTHER SYSTEMS

Номер: US20140284001A1
Автор: Amstad Esther, HOLTZE Christian, Weitz David A.
Принадлежит:

The present invention generally relates to microfluidics, and to spray drying and other drying techniques. By at least partially drying fluids within a microfluidic channel, instead of or in addition to conventional spray drying techniques, better control of the drying process can be achieved in certain aspects of the invention. In addition, various embodiments of the invention are generally directed to systems and methods for drying fluids contained within a channel such as a microfluidic channel. For example, a fluid may be partially or completely dried within a microfluidic channel, prior to being sprayed into a collection region. In some embodiments, gases such as air may be directed into a channel containing a fluid, which may facilitate drying of the fluid. In some cases, the fluid may be accelerated due to the introduction of gases into the channel, and in certain embodiments, droplets of fluid may be disrupted to form smaller droplets as a result. In certain cases, the fluids may also be dried to form supersaturated droplets. 1. A spray dryer for use in drying liquids , comprising: a first microfluidic channel;', 'second and third microfluidic channels each intersecting the first microfluidic channel at substantially non-right angles at a first intersection; and', 'fourth and fifth microfluidic channels each intersecting the first microfluidic channel at substantially non-right angles at a second intersection; and, 'an article comprisinga collection region that receives output from the first microfluidic channel.2. The spray dryer of claim 1 , wherein the second intersection is downstream of the first intersection.3. The spray dryer of any one of or claim 1 , wherein the substantially non-right angles at the first intersection are each obtuse angles.43. The spray dryer of any one of - claims 1 , wherein the substantially non-right angles at the first intersection are each about 135°.54. The spray dryer of any one of - claims 1 , wherein the substantially non ...

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Номер записи: 81
14-07-2016 дата публикации

DETERMINATION OF IMMUNE CELLS AND OTHER CELLS

Номер: US20160201129A1
Автор: Gilbert John R., Heyman John, Weitz David A.
Принадлежит: President and Fellows of Harvard College

The present invention generally relates to fluidic droplets, and to techniques for screening or sorting such fluidic droplets. In some embodiments, the fluidic droplets may contain cells such as immune cells, which can be analyzed to determine receptor sequences or other useful properties of the cells. For example, in one aspect, the present invention is generally related to determining immune cell receptors by encapsulating immune cells and target cells in microfluidic droplets, determining the effect of the immune cells on the target cells, and for those immune cells that kill or otherwise adversely affect the target cells, determining one or more receptor sequences of those immune cells. The target cells may be, for example, cancer cells or virally-infected cells. In some cases, the receptor sequences can be used, for example, to identify certain properties of the immune cells, to screen for drugs or other therapeutic agents, or the like. 1. A method of determining immune cell receptors , the method comprising:encapsulating immune cells and target cells in microfluidic droplets contained within a microfluidic channel such that at least some of the microfluidic droplets contain at least one immune cell and at least one target cell;determining viability of the target cell after exposure of the target cell to the immune cell;separating the microfluidic droplets on the basis of the viability of the target cell; andfor the microfluidic droplets containing at least one immune cell and at least one non-viable target cell, determining a receptor sequence of the at least one immune cell.2. The method of any claim 1 , wherein the immune cells comprise T-cells.3. The method of any one of or claim 1 , wherein the immune cells comprise CD8+ T-cells.4. The method of any one of - claim 1 , wherein at least about 90% of the immune cells are CD8+ T-cells.5. The method of any one of - claim 1 , wherein the immune cells comprise B-cells.6. The method of any one of - claim 1 , ...

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Номер записи: 82
13-07-2017 дата публикации

HYDROGEL COMPOSITIONS COMPRISING ENCAPSULATED CELLS AND METHODS OF USE THEREOF

Номер: US20170196818A1
Автор: Mao Angelo S., Mooney David J., SHIN JAE-WON, Utech Stefanie, Uzun Oktay, Weitz David A.
Принадлежит:

The present invention provides injectable compositions comprising cells encapsulated in hydrogel capsules and methods of preparing these compositions. The present invention also provides methods for using these compositions to promote hematopoiesis and to treat or prevent cardiovascular and immunological disorders in a subject. 1. A composition comprising a plurality of hydrogel capsules , whereinat least 90% of said hydrogel capsules in said composition comprise a cell and a hydrogel encapsulating said cell, wherein said hydrogel encapsulating said cell has a thickness of less than 20 microns.2. The composition of claim 1 , wherein at least 70% of said hydrogel capsules comprise a single cell.3. The composition of claim 1 , wherein the cell is a mesenchymal stem cell (MSC) or a progenitor thereof claim 1 , a hematopoietic stem cell (HSC) or a progenitor thereof claim 1 , or an endothelial progenitor cell.4. (canceled)5. The composition of claim 1 , wherein the hydrogel comprises at least one polymer selected from the group consisting of alginate claim 1 , agarose claim 1 , poly(ethylene glycol dimethacrylate) claim 1 , polylactic acid claim 1 , polyglycolic acid claim 1 , PLGA claim 1 , gelatin claim 1 , collagen claim 1 , agarose claim 1 , pectin claim 1 , poly(lysine) claim 1 , polyhydroxybutyrate claim 1 , poly-epsilon-caprolactone claim 1 , polyphosphazines claim 1 , poly(vinyl alcohol) claim 1 , poly(alkylene oxide) claim 1 , poly(ethylene oxide) claim 1 , poly(allylamine) claim 1 , poly(acrylate) claim 1 , poly(4-aminomethylstyrene) claim 1 , pluronic polyol claim 1 , polyoxamer claim 1 , poly(uronic acid) claim 1 , poly(anhydride) and poly(vinylpyrrolidone).6. (canceled)7. The composition of claim 5 , wherein the polymer is alginate.8. The composition of claim 5 , wherein the polymer comprises polymer chains cross-linked to each other using a divalent or trivalent cation selected from the group consisting of Ca claim 5 , Mg claim 5 , Sr claim 5 , Ba claim 5 ...

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Номер записи: 83
29-07-2021 дата публикации

DROPLET CREATION TECHNIQUES

Номер: US20210229099A1
Автор: Abate Adam R., Weitz David A.
Принадлежит: President and Fellows of Harvard College

The present invention is generally related to systems and methods for producing droplets. The droplets may contain varying species, e.g., for use as a library. In some cases, at least one droplet is used to create a plurality of droplets, using techniques such as flow-focusing techniques. In one set of embodiments, a plurality of droplets, containing varying species, can be divided to form a collection of droplets containing the various species therein. A collection of droplets, according to certain embodiments, may contain various subpopulations of droplets that all contain the same species therein. Such a collection of droplets may be used as a library in some cases, or may be used for other purposes. 120-. (canceled)21. A method for forming a plurality of divided droplets , comprising:providing a plurality of droplets, each droplet comprising a first fluid surrounded by a second fluid; andpassing at least some of the droplets through a droplet maker such that each of the droplets passing through the microfluidic channel are divided by the droplet maker to form a collection of divided droplets,wherein the collection comprises a plurality of groups of divided droplets, where the divided droplets within each group are substantially indistinguishable but each group of divided droplets is distinguishable from the other groups of divided droplets.22. The method of claim 21 , wherein the droplet maker is a T-junction droplet maker.23. The method of claim 21 , wherein the droplet maker is a micro-capillary droplet maker claim 21 , preferably a co-flow micro-capillary droplet maker.24. The method of claim 21 , wherein the first fluid and the second fluid are immiscible.25. The method of claim 21 , wherein the collection of divided droplets is surrounded by the second fluid.26. The method of claim 21 , wherein each group of divided droplets is distinguishable from the other groups of divided droplets due to different species contained within each group of droplets or on ...

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Номер записи: 84
06-08-2015 дата публикации

FLUOROCARBON EMULSION STABILIZING SURFACTANTS

Номер: US20150217246A1
Автор: HOLTZE Christian, Hutchison John Brian, Weitz David A.
Принадлежит:

Surfactants (e.g., fluorosurfactants) for stabilizing aqueous or hydrocarbon droplets in a fluorophilic continuous phase are presented. In some embodiments, fluorosurfactants include a fluorophilic tail soluble in a fluorophilic (e.g., fluorocarbon) continuous phase, and a headgroup soluble in either an aqueous phase or a lipophilic (e.g., hydrocarbon) phase. The combination of a fluorophilic tail and a headgroup may be chosen so as to create a surfactant with a suitable geometry for forming stabilized reverse emulsion droplets having a disperse aqueous or lipophilic phase in a continuous, fluorophilic phase. In some embodiments, the headgroup is preferably non-ionic and can prevent or limit the adsorption of molecules at the interface between the surfactant and the discontinuous phase. This configuration can allow the droplet to serve, for example, as a reaction site for certain chemical and/or biological reactions. In another embodiment, aqueous droplets are stabilized in a fluorocarbon phase at least in part by the electrostatic attraction of two oppositely charged or polar components, one of which is at least partially soluble in the dispersed phase, the other at least partially soluble in the continuous phase. One component may provide colloidal stability of the emulsion, and the other may prevent the adsorption of biomolecules at the interface between a component and the discontinuous phase. Advantageously, surfactants and surfactant combinations of the invention may provide sufficient stabilization against coalescence of droplets, without interfering with processes that can be carried out inside the droplets. 1105-. (canceled)106. A surfactant , comprising a block copolymer including a perfluorinated polyether (PFPE) block coupled to a polyethylene glycol (PEG) block via an amide bond , and comprising a formula —(CFO)—(CF)—CONH— wherein n , m , x , and y are positive integers.107. The surfactant of claim 106 , wherein the surfactant comprises one block of ...

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Номер записи: 85
25-06-2020 дата публикации

POLY(ACID) MICROCAPSULES AND RELATED METHODS

Номер: US20200197894A1
Автор: Deverney Brendon, Nawar Sara, Weitz David A., WERNER Joerg G.
Принадлежит: PRESIDENT AND FELLOW OF HARVARD COLLEGE

Microcapsules and techniques for the formation of microcapsules are generally described. In some embodiments, the microcapsules are formed in an emulsion (e.g., a multiple emulsion). In some embodiments, the microcapsule may be suspended in a carrying fluid containing the microcapsule that, in turn, contain the smaller droplets. In some embodiments, the microcapsules comprise a shell and a droplet at least partially contained within the shell (e.g., encapsulated within the shell), and may be suspended in a carrier fluid. In certain embodiments, the shell is a hydrogel comprising a poly(acid). In some cases, the poly(acid) is a polyanion. In some cases, the shell does not comprise a poly(base) or polycation (e.g., a polycationic poly electrolyte). In some embodiments, the microcapsules comprise a shell comprising a poly(acid) and a poly(anhydride). 1. A method , comprising:forming a microfluidic droplet comprising a first fluid contained within a carrying fluid, the first fluid comprising an anhydride;polymerizing some of the anhydride within the microfluidic droplet to form a poly(anhydride) to cause the droplet to form a microcapsule;cross-linking the poly(anhydride) within the microcapsule; andhydrolyzing some of the anhydride within the microcapsule to form carboxylic acid.2. The method of claim 1 , wherein the poly(anhydride) comprises methacrylic anhydride.3. The method of any one of or claim 1 , wherein the poly(anhydride) comprises pentenoic anhydride.4. The method of any one of - claim 1 , wherein polymerizing some of the anhydride comprises exposing the anhydride to UV light.5. The method of any one of - claim 1 , wherein polymerizing some of the anhydride comprises exposing the anhydride to a photoinitiator.6. The method of any one of - claim 1 , wherein the microfluidic droplet has an average cross-sectional diameter of greater than or equal to 15 micrometers.7. The method of any one of - claim 1 , wherein the microfluidic droplet has an average cross- ...

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Номер записи: 86
09-10-2014 дата публикации

Particle-assisted nucleic acid sequencing

Номер: US20140303039A1
Автор: Adam R. Abate, David A. Weitz
Принадлежит: Harvard College

This invention generally relates to particle-assisted nucleic acid sequencing. In some embodiments, sequencing may be performed in a microfluidic device, which can offer desirable properties, for example, minimal use of reagents, facile scale-up, and/or high throughput. In one embodiment, a target nucleic acid may be exposed to particles having nucleic acid probes. By determining the binding of the particles to the target nucleic acid, the sequence of the target nucleic acid (or at least a portion of the target nucleic acid) can be determined. The target nucleic acid may be encapsulated within a fluidic droplet with the particles having nucleic acid probes, in certain instances. In some cases, the sequence of the target nucleic acid may be determined, based on binding of the particles, using sequencing by hybridization (SBH) algorithms or other known techniques.

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Номер записи: 87
16-10-2014 дата публикации

ELECTRONIC CONTROL OF FLUIDIC SPECIES

Номер: US20140305799A1
Автор: Ahn Keunho, Cheng Zhengdong, Cristobal-Azkarate Galder, Link Darren Roy, Weitz David A.
Принадлежит: President and Fellows of Harvard College

Various aspects of the present invention relate to the control and manipulation of fluidic species, for example, in microfluidic systems. In one aspect, the invention relates to systems and methods for making droplets of fluid surrounded by a liquid, using, for example, electric fields, mechanical alterations, the addition of an intervening fluid, etc. 1273-. (canceled)274. A method of introducing a fluid into a droplet , comprising:providing a droplet in a carrier fluid within a microchannel; andcontacting the droplet with a fluid stream, thereby introducing a fluid into the droplet.275. The method of claim 274 , wherein the droplet and the fluid stream comprise hydrophilic liquids.276. The method of claim 275 , wherein the droplet and the fluid stream comprise aqueous solutions.277. The method of claim 274 , wherein the carrier fluid comprises a hydrophobic liquid.278. The method of claim 274 , wherein the droplet and the fluid stream comprise a chemical claim 274 , biological claim 274 , or biochemical entity.279. The method of claim 278 , wherein the droplet and the fluid stream comprise different chemical claim 278 , biological claim 278 , or biochemical entities.280. The method of claim 274 , wherein the microchannel has a maximum cross-sectional dimension of less than about 500 micrometers.281. The method of claim 274 , further comprising mixing the fluid introduced into the droplet with the contents of the droplet.282. The method of claim 274 , further comprising:providing a sensor for sensing a characteristic of the droplet after fluid has been introduced into the droplet, wherein the characteristic is fluorescence, spectroscopic signature, radioactivity, mass, volume, density, temperature, viscosity, pH, or concentration of a substance.283. The method of claim 282 , wherein the characteristic is the presence of a nucleic acid.284. The method of claim 274 , wherein the droplet flows into an intersection comprising the microchannel and a second fluid ...

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Номер записи: 88
02-08-2018 дата публикации

MICROPARTICLES, METHODS FOR THEIR PREPARATION AND USE

Номер: US20180214385A1
Автор: Amstad Esther, Huanan Wang, Mao Angelo S., Mooney David J., Ostafe Raluca, Prodanovich Radivoje, Utech Stefanie, Weitz David A., Wilking Connie Chang
Принадлежит: President and Fellows of Harvard College

The invention relates to microparticles comprising a crosslinked gel and methods for making and using same. 142-. (cancelled)43. A method of forming microparticles , comprising:passing a plurality of microdroplets comprising a gel precursor through a curved microfluidic channel having at least one dimension measuring from about 5 μm to about 200 μm; andcrosslinking the gel precursor to form a plurality of microparticles comprising a crosslinked gel, the microparticles having a shape conforming to the shape of the curved microfluidic channel.44. The method of claim 43 , wherein the microparticles have a coefficient of variation in size distribution of from about 0.03 to about 0.05.45. The method of claim 43 , wherein the microparticles are crescent- or hook-shaped.46. The method of claim 43 , wherein crosslinking the gel precursor to form a plurality of microparticles comprises microfluidically forming the microdroplets.47. The method of claim 43 , wherein the gel precursor comprises one or more crosslinkable linear polysaccharides and one or more crosslinking agents.48. The method of claim 47 , comprising crosslinking the gel precursor with a crosslinking promoter to promote crosslinking of the one or more crosslinkable linear polysaccharides.49. The method of claim 48 , wherein the crosslinking agent does not substantially crosslink the one or more crosslinkable linear polysaccharides in an initial state claim 48 , but upon contacting with the crosslinking promoter claim 48 , crosslinks the one or more crosslinkable linear polysaccharides in a second state.50. The method of claim 49 , wherein the initial state comprises a sequestered state of the one or more cros slinking agents and the second state comprises an unsequestered state of the one or more crosslinking agents.51. The method of claim 48 , wherein said cros slinking promoter comprises a change in pH claim 48 , a change in temperature claim 48 , a change in ionic strength or combinations thereof.52. The ...

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Номер записи: 89
11-07-2019 дата публикации

ACOUSTIC WAVES IN MICROFLUIDICS

Номер: US20190211293A1
Автор: Abate Adam R., Agresti Jeremy, Franke Thomas, Schmid Lothar, Weitz David A., Wixforth Achim
Принадлежит:

Various aspects of the present invention relate to the control and manipulation of fluidic species, for example, in microfluidic systems. In one set of embodiments, droplets may be sorted using surface acoustic waves. The droplets may contain cells or other species. In some cases, the surface acoustic waves may be created using a surface acoustic wave generator such as an interdigitated transducer, and/or a material such as a piezoelectric substrate. The piezoelectric substrate may be isolated from the microfluidic substrate except at or proximate the location where the droplets are sorted, e.g., into first or second microfluidic channels. At such locations, the microfluidic substrate may be coupled to the piezoelectric substrate (or other material) by one or more coupling regions. In some cases, relatively high sorting rates may be achieved, e.g., at rates of at least about 1,000 Hz, at least about 10,000 Hz, or at least about 100,000 Hz, and in some embodiments, with high cell viability after sorting. 19-. (canceled)10. A method , comprising:directing a plurality of droplets through an inlet microfluidic channel to a single junction between the inlet microfluidic channel, a first channel, a second channel, and a third channel; andapplying a first surface acoustic wave to some of the droplets to cause the droplets to enter the first channel and applying a second surface acoustic wave to some of the droplets to cause the droplets to enter the second channel, wherein in the absence of the first or second surface acoustic waves, the droplets enter the third channel.11. The method of claim 10 , wherein the first surface acoustic wave and/or the second surface acoustic wave has an average frequency of between about 130 MHz and about 160 MHz.12. The method of claim 10 , wherein the first surface acoustic wave and/or the second surface acoustic wave has an average frequency of between about 140 MHz and about 150 MHz.13. A method claim 10 , comprising:directing a plurality ...

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Номер записи: 90
09-08-2018 дата публикации

POLYMERSOMES, LIPOSOMES, AND OTHER SPECIES ASSOCIATED WITH FLUIDIC DROPLETS

Номер: US20180221491A1
Автор: KIM Jin-Woong, Lee Daeyeon, Shum Ho Cheung, Weitz David A., Yoon Insun
Принадлежит:

The present invention relates generally to vesicles such as liposomes, colloidosomes, and polymersomes, as well as techniques for making and using such vesicles. In some cases, the vesicles may be at least partially biocompatible and/or biodegradable. The vesicles may be formed, according to one aspect, by forming a multiple emulsion comprising a first droplet surrounded by a second droplet, which in turn is surrounded by a third fluid, where the second droplet comprises lipids and/or polymers, and removing fluid from the second droplet, e.g., through evaporation or diffusion, until a vesicle is formed. In certain aspects, the size of the vesicle may be controlled, e.g., through osmolarity, and in certain embodiments, the vesicle may be ruptured through a change in osmolarity. In some cases, the vesicle may contain other species, such as fluorescent molecules, microparticles, pharmaceutical agents, etc., which may be released upon rupture. Yet other aspects of the invention are generally directed to methods of making such vesicles, kits involving such vesicles, or the like. 127-. (canceled)28. A method of forming a polymersome comprising a species encapsulated therein , comprising:(a) generating a double emulsion comprising an outer phase substantially immiscible with a middle phase, which middle phase is in turn substantially immiscible with an inner phase, wherein said inner phase comprises a species, and wherein said middle phase comprises an amphiphilic diblock copolymer in a solvent, wherein the amphiphilic diblock copolymer comprising hydrophilic and hydrophobic blocks; and 'wherein a molecular weight ratio of said hydrophilic to hydrophobic blocks is selected such that said polymer membrane is degradable upon application of an osmotic pressure shock.', '(b) removing said solvent of said middle phase to form a polymer membrane, thereby yielding said polymersome comprising said species encapsulated therein,'}29. The method of claim 28 , wherein said outer phase ...

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Номер записи: 91
10-08-2017 дата публикации

Microcapsules and uses thereof

Номер: US20170224849A1
Автор: David A. Weitz, Joseph D. Brain, Maximilian ZIERINGER, Nagarjun Konduru Vendata, Nichlaus James Carroll, Rajiv Gupta, Ramon Molina
Принадлежит: General Hospital Corp, Harvard College

Certain aspects of the present invention relates to microcapsules comprising a core; and a hydrophobic, cross-linked polymeric shell, as well as method for making and using same. Some embodiments of the present invention relate to microcapsules comprising a core; and a hydrophobic, cross-linked polymeric shell. These microcapsules can be used in a variety of applications, including agriculture, encapsulation of food ingredients, health care, cosmetics (e.g., perfumes, detergents, and sunscreen), coatings (e.g., paints and pigments), additives, catalysis, and oil recovery.

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Номер записи: 92
10-08-2017 дата публикации

MICROFLUIDIC DEVICE FOR STORAGE AND WELL-DEFINED ARRANGEMENT OF DROPLETS

Номер: US20170225167A1
Автор: Agresti Jeremy, BOEHM Christian, Koester Sarah, Rowat Amy, Weitz David A.
Принадлежит:

The present invention relates to systems and methods for the arrangement of droplets in pre-determined locations. Many applications require the collection of time-resolved data. Examples include the screening of cells based on their growth characteristics or the observation of enzymatic reactions. The present invention provides a tool and related techniques which addresses this need, and which can be used in many other situations. The invention provides, in one aspect, a tool that allows for stable storage and indexing of individual droplets. The invention can interface not only with microfluidic/microscale equipment, but with macroscopic equipment to allow for the easy injection of liquids and extraction of sample droplets, etc. 155-. (canceled)56. A device for arranging droplets comprising:a channel comprising a series of pots connected via a series of constrictions arranged such that each pot is in direct fluid communication with no more than two other pots and each pot is connected to exactly two constrictions.57. The device of claim 56 , further comprising a plurality of channels each comprising a series of pots connected via a series of constrictions arranged such that each pot is in direct fluid communication with no more than two other pots and each pot is connected to exactly two constrictions.5861-. (canceled)62. The device of claim 57 , wherein the volumes of the pots are about 10 nanoliters.64. The device of claim 56 , further comprising:a gas-permeable membrane that separates the contents of the pot from ambient atmosphere.6571-. (canceled) This application claims the benefit of U.S. Provisional Patent Application Ser. No. 61/048,304, filed Apr. 28, 2008, entitled “Microfluidic Device for Storage and Well-Defined Arrangement of Droplets,” by Weitz, et al., the entirety of which is incorporated herein by reference.The present invention relates generally to systems and methods for the arrangement of droplets in pre-determined locations.Droplet-based ...

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Номер записи: 93
09-07-2020 дата публикации

POLYMERSOMES, COLLOIDOSOMES, LIPOSOMES, AND OTHER SPECIES ASSOCIATED WITH FLUIDIC DROPLETS

Номер: US20200215193A1
Автор: KIM Jin-Woong, Lee Daeyeon, Shum Ho Cheung, Weitz David A., Yoon Insun
Принадлежит: President and Fellows of Harvard College

The present invention relates generally to vesicles such as liposomes, colloidosomes, and polymersomes, as well as techniques for making and using such vesicles. In some cases, the vesicles may be at least partially biocompatible and/or biodegradable. The vesicles may be formed, according to one aspect, by forming a multiple emulsion comprising a first droplet surrounded by a second droplet, which in turn is surrounded by a third fluid, where the second droplet comprises lipids and/or polymers, and removing fluid from the second droplet, e.g., through evaporation or diffusion, until a vesicle is formed. In certain aspects, the size of the vesicle may be controlled, e.g., through osmolarity, and in certain embodiments, the vesicle may be ruptured through a change in osmolarity. In some cases, the vesicle may contain other species, such as fluorescent molecules, microparticles, pharmaceutical agents, etc., which may be released upon rupture. Yet other aspects of the invention are generally directed to methods of making such vesicles, kits involving such vesicles, or the like. 127-. (canceled)28. A method of forming a polymersome comprising a species encapsulated therein , said method comprising:a. generating a double emulsion comprising an outer phase substantially immiscible with a middle phase, which middle phase is in turn substantially immiscible with an inner phase, wherein said inner phase comprises said species, and wherein said middle phase comprises an amphiphilic diblock copolymer in a solvent, wherein said amphiphilic diblock copolymer comprises hydrophilic and hydrophobic blocks; and wherein said removing comprises said solvent dewetting from said inner phase, wherein sad solvent comprises a mixture of a first fluid and a second fluid, wherein volume fractions of said first fluid and said second fluid in said mixture are selected such that said dewetting yields said polymerosome, and', 'wherein a molecular weight ratio of said hydrophilic to hydrophobic ...

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Номер записи: 94
19-08-2021 дата публикации

DETERMINATION OF CELLS USING AMPLIFICATION

Номер: US20210254129A1
Автор: Weitz David A., Zhang Huidan
Принадлежит: President and Fellows of Harvard College

The present invention generally relates to microfluidics and, in particular, to systems and methods for determining cells using amplification. In one set of embodiments, cells are encapsulated within droplets and nucleic acids from the cells amplified within the droplets. The droplets may then be pooled together and the amplified nucleic acids can be determined using PCR or other suitable techniques. In some embodiments, techniques such as these can be used to detect relatively rare cells that may be present, e.g., if the droplets are amplified using conditions able to selectively amplify nucleic acids arising from the relatively rare cells. 150-. (canceled)51. A method , comprising:within a plurality of droplets encapsulating nucleic acids arising from a plurality of cells, applying conditions able to selectively amplify a target nucleic acid sequence suspected of being present within the nucleic acids;combining the interiors of the droplets together to form a combined fluid; anddetermining amplified nucleic acids contained within the combined fluid.52. A method , comprising:encapsulating cells within a plurality of droplets;lysing at least some of the cells within the plurality of droplets to release nucleic acids from the cells into the interior of the droplets;within the interior of the droplets, applying conditions able to selectively amplify a target nucleic acid sequence suspected of being present within the nucleic acids released from the cells; anddetermining amplified nucleic acids contained within the droplets.53. The method of claim 51 , wherein the target nucleic acid sequence is a DNA sequence.54. The method of claim 51 , wherein the target nucleic acid sequence is an RNA sequence.55. The method of claim 51 , wherein the target nucleic acid sequence is an mRNA sequence.56. The method of claim 51 , wherein applying conditions able to selectively amplify a target nucleic acid sequence comprises applying conditions able to cause amplification via PCR.57. ...

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Номер записи: 95
01-09-2016 дата публикации

VALVES AND OTHER FLOW CONTROL IN FLUIDIC SYSTEMS INCLUDING MICROFLUIDIC SYSTEMS

Номер: US20160250638A1
Автор: Abate Adam R., Weitz David A.
Принадлежит: President and Fellows of Harvard College

Articles and methods for controlling flow in fluidic systems, especially in microfluidic systems, are provided. In one aspect, a microfluidic system described herein includes a configuration such that the actuation of a single valve can allow the switching of fluids from a first fluid path (e.g., a first channel section) to a second fluid path (e.g., a second channel section). This may be achieved, for example, by incorporating a valve with a first channel section, which may have a lower hydrodynamic resistance than a second channel section prior to actuation of the valve. Actuation of the valve can cause only the hydrodynamic resistance of the first channel section to increase, thereby redirecting fluid flow into the second channel section (which now has a relatively lower hydrodynamic resistance). In some embodiments, the valve comprises a control channel for introducing a positive or reduced pressure, and is adapted to modulate fluid flow in an adjacent channel section by constricting or expanding the channel section. For example, the valve and/or the channel section may be formed in a flexible material and actuation of the valve may be achieved by applying a positive or reduced pressure to the valve to cause deformation of both the valve and the channel section. Another aspect of the invention includes articles and methods associated with manipulation of multiphase materials (e.g., dispersions). For instance, one or more valves may be combined with a flow focusing system so as to form droplets of different volumes and/or frequencies without the need to vary flow rates of the fluids when they are introduced into the fluidic system. 126-. (canceled)27. A method , comprising:without the need to change a cross-sectional dimension of a channel of a microfluidic system during use, producing in the microfluidic system a series of single, substantially uniform droplets having a first volume and comprising a subject fluid, the droplets being surrounded by a continuous ...

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Номер записи: 96
13-11-2014 дата публикации

Deformable Platforms for Biological Assays

Номер: US20140336072A1
Автор: Butler James, Ehrlicher Allen, Fredberg Jeffrey J., Krishnan Ramaswamy, PARK Chan Young, Weitz David A.
Принадлежит:

A platform for biological assays includes a base substrate providing structural support to the platform, at least one surface of the base substrate coated with position markers, a first deformable layer positioned on top of the base substrate, and a second deformable layer positioned on top of the first deformable layer, the second deformable layer embedded with deformation markers. 1. A platform for biological assays , comprising:a base substrate providing structural support to the platform, at least one surface of the base substrate coated with position markers;a first deformable layer positioned on top of the base substrate; anda second deformable layer positioned on top of the first deformable layer, the second deformable layer embedded with deformation markers.2. The platform for biological assays in claim 1 , wherein the first and second deformable layers are made of a primarily same material and wherein the first and second deformation layers are combined and merged into an indistinguishable layer with deformation markers embedded near an outer surface of the indistinguishable layer.3. The platform for biological assays in claim 1 , wherein the stiffness of at least one of the first and second deformable layers is tunable.4. The platform for biological assays in claim 1 , wherein at least one of the first and second deformable layers is primarily made of polydimethylsiloxane (PDMS).5. The platform for biological assays in claim 1 , wherein at least one of the first and second deformable layers has a stiffness less than 10 kilopascal (kPa).6. The platform for biological assays in claim 1 , wherein the position markers and the deformation markers are fluorescent beads.7. The platform for biological assays in claim 6 , wherein the position markers and the deformation markers have different fluorescent colors.8. The platform for biological assays in claim 1 , wherein the first deformable layer has a thickness of about 50-300 μm and the second deformable layer has ...

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Номер записи: 97
31-08-2017 дата публикации

FLUID INJECTION USING ACOUSTIC WAVES

Номер: US20170246634A1
Автор: Franke Thomas, Weitz David A.
Принадлежит:

The present invention generally relates to the manipulation of fluids using acoustic waves such as surface acoustic waves. In some aspects, one fluid may be introduced into another fluid via application of suitable acoustic waves. For example, a fluid may be added or injected into another fluid by applying acoustic waves where, in the absence of the acoustic waves, the fluid cannot be added or injected, e.g., due to the interface or surface tension between the fluids. Thus, for example, a fluid may be injected into a droplet of another fluid. Other embodiments of the invention are generally directed to systems and methods for making or using such systems, kits involving such systems, or the like. 1. A method , comprising:providing a microfluidic system comprising a first microfluidic channel and a second microfluidic channel contacting the first microfluidic channel at a junction;providing a droplet of the first fluid contained by a carrying fluid in the first microfluidic channel, and a second fluid in the second microfluidic channel, wherein the first fluid and the second fluid contact each other at least partially within the junction to define a fluidic interface; andapplying acoustic waves to the interface to urge the second fluid to flow into the droplet wherein, in the absence of the acoustic waves, the second fluid is not urged to enter the droplet.2. The method of claim 1 , wherein the acoustic waves are applied to at least a portion of the junction.3. The method of any one of or claim 1 , wherein the acoustic waves have an amplitude whose magnitude varies in time.4. The method of any one of - claim 1 , wherein the acoustic waves are applied to the first fluid in a direction of flow of the first fluid.5. The method of any one of - claim 1 , wherein the acoustic waves are applied to the first fluid opposite a direction of flow of the first fluid.6. The method of any one of - claim 1 , wherein the acoustic waves are applied to the second fluid in a direction ...

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Номер записи: 98
20-11-2014 дата публикации

SYSTEMS AND METHODS FOR DROPLET PRODUCTION AND/OR FLUIDIC MANIPULATION

Номер: US20140338753A1
Автор: Abate Adam R., Sperling Ralph Alexander, Weitz David A.
Принадлежит:

The present invention generally relates to systems and techniques for manipulating fluids and/or making droplets. In certain aspects, the present invention generally relates to droplet production. The droplets may be formed from fluids from different sources. In one set of embodiments, the present invention is directed to a microfluidic device comprising a plurality of droplet-making units, and/or other fluidic units, which may be substantially identical in some cases. Substantially each of the fluidic units may be in fluidic communication with a different source of a first fluid and a common source of a second fluid, in certain embodiments. In one aspect, substantially the same pressure may be applied to substantially all of the different sources of fluid, which may be used to cause fluid to move from the different sources into the microfluidic device. In some cases, the fluids may interact within the fluidic units, e.g., by reacting, or for the production of droplets within the microfluidic device. In some cases, the droplets may be used, for example, to form a library of droplets. 1. A method , comprising:providing a microfluidic device comprising a plurality of droplet-making units, substantially each of the droplet-making units in fluidic communication with a different source of droplet fluid and a common source of carrier fluid;applying substantially the same pressure and/or pressure drop to substantially all of the different sources of droplet fluid to cause droplet fluid to move from the different sources of droplet fluid into the microfluidic device; andproducing, within the microfluidic device, a set comprising a plurality of droplets, substantially each droplet of the set comprising droplet fluid from only one of the different sources of droplet fluid, the plurality of droplets of the set being contained within carrier fluid from the common source of carrier fluid.2. The method of claim 1 , wherein applying substantially the same pressure and/or pressure ...

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Номер записи: 99
13-09-2018 дата публикации

ACOUSTIC WAVE SORTING

Номер: US20180257076A1
Автор: Franke Thomas, Ung Warren Lloyd, Weitz David A.
Принадлежит: President and Fellows of Harvard College

The present invention generally relates to the manipulation of species using acoustic waves such as surface acoustic waves. In some aspects, a channel such as a microfluidic channel may be provided having two or more outlets, and acoustic waves applied to species within the channel to determine which outlet the species is directed to. For instance, surface acoustic waves may be applied to a species such as a cell or a particle to deflect it from the channel into a groove or other portion that directs it to a different outlet. In some cases, surprisingly, this deflection of species may be in a different direction than the incident acoustic waves on the channel. Other embodiments of the present invention are generally directed to kits including such systems, techniques for producing such systems, or the like. 144-. (canceled)45. An apparatus , comprising:a microfluidic channel having a groove defined in a first surface, the groove having a cross-sectional dimension sized to allow a mammalian cell to fit within the groove; andan acoustic wave generator positioned proximate a second surface of the microfluidic channel, the second surface adjacent to the first surface.46. The apparatus of claim 45 , wherein the microfluidic channel comprises an inlet and a first outlet claim 45 , and the groove directs the species to a second outlet.47. The apparatus of claim 45 , wherein the groove is positioned at a nonzero angle with respect to the microfluidic channel.48. The apparatus of claim 45 , wherein the groove is defined by at least one wall that is not parallel or perpendicular to the first surface of the microfluidic channel.49. The apparatus of claim 45 , wherein the groove has a cross-sectional dimension of at least about 20 micrometers.50. The apparatus of claim 45 , wherein the groove has a cross-sectional dimension that is less than a cross-sectional dimension of the microfluidic channel adjacent the groove.51. The apparatus of claim 50 , wherein the acoustic wave ...

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Номер записи: 100