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    ELECTRONIC CONTROL OF FLUIDIC SPECIES
    51.
    发明申请
    ELECTRONIC CONTROL OF FLUIDIC SPECIES 审中-公开

    公开(公告)号：US20200276578A1

    公开(公告)日：2020-09-03

    申请号：US16813106

    申请日：2020-03-09

    Applicant: President and Fellows of Harvard College

    Inventor： David A. Weitz ,  Darren Roy Link ,  Galder Cristobal-Azkarate ,  Zhengdong Cheng ,  Keunho Ahn

    IPC: B01L3/00 ,  B01F5/06 ,  B01F13/00 ,  B01J19/00 ,  G01N15/14 ,  G01N15/10

    Abstract: 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.

    Particle-assisted nucleic acid sequencing
    53.
    发明授权
    Particle-assisted nucleic acid sequencing 有权

    公开(公告)号：US10457977B2

    公开(公告)日：2019-10-29

    申请号：US15427511

    申请日：2017-02-08

    Applicant: President and Fellows of Harvard College

    Inventor： David A. Weitz ,  Adam R. Abate

    IPC: C12Q1/68 ,  C12Q1/6809 ,  C12Q1/6869 ,  B01L3/00 ,  C12Q1/6834 ,  C12Q1/6874

    Abstract: 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.

    MICROFLUIDIC SEQUENCING TECHNIQUES
    57.
    发明申请
    MICROFLUIDIC SEQUENCING TECHNIQUES 审中-公开

    公开(公告)号：US20190185800A1

    公开(公告)日：2019-06-20

    申请号：US16319196

    申请日：2017-07-20

    Applicant: President and Fellows of Harvard College

    Inventor： David A. Weitz ,  Huidan Zhang ,  Nai Wen Cui

    IPC: C12M3/06 ,  C12Q1/6858 ,  C12M1/00 ,  C12Q1/6806

    CPC classification number: C12M23/16 ,  C12M3/00 ,  C12M47/06 ,  C12Q1/6806 ,  C12Q1/6858 ,  C12Q2521/301 ,  C12Q2525/131 ,  C12Q2535/122 ,  C12Q2563/159 ,  C12Q2565/629

    Abstract: 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.

    Formation and control of fluidic species
    58.
    发明授权
    Formation and control of fluidic species 有权

    公开(公告)号：US10293341B2

    公开(公告)日：2019-05-21

    申请号：US14662668

    申请日：2015-03-19

    Applicant: President and Fellows of Harvard College

    Inventor： Darren Roy Link ,  David A. Weitz ,  Manuel Marquez-Sanchez ,  Zhengdong Cheng

    IPC: B05B5/03 ,  B01F5/06 ,  B01J2/04 ,  B01L3/00 ,  B05B7/06 ,  B01F13/00

    Abstract: 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.

    Assays and other reactions involving droplets
    60.
    发明授权
    Assays and other reactions involving droplets 有权

    公开(公告)号：US10221437B2

    公开(公告)日：2019-03-05

    申请号：US15884215

    申请日：2018-01-30

    Applicant: President and Fellows of Harvard College

    Inventor： David A. Weitz ,  Jeremy Agresti ,  Liang-Yin Chu ,  Jin-Woong Kim ,  Amy Rowat ,  Morten Sommer ,  Gautam Dantas ,  George Church

    IPC: C12P19/34 ,  C12Q1/68 ,  B01F13/00 ,  B01F3/08 ,  B01J13/00 ,  C12Q1/686 ,  C12Q1/6848 ,  C12Q1/6834 ,  G01N15/14 ,  B01L3/00 ,  G01N15/10

    Abstract: 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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