公开(公告)号：US11618024B2

公开(公告)日：2023-04-04

申请号：US17184202

申请日：2021-02-24

Applicant: President and Fellows of Harvard College ,  Brandeis University

Inventor： Seth Fraden ,  Hakim Boukellal ,  Yanwei Jia ,  Seila Selimovic ,  Amy Rowat ,  Jeremy Agresti ,  David A. Weitz

IPC: G01N1/28 ,  G01N15/00 ,  B01L3/00 ,  G01N15/02 ,  G01N15/14 ,  F17D1/12

Abstract: 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.

公开(公告)号：US20230045126A1

公开(公告)日：2023-02-09

申请号：US17790450

申请日：2021-01-13

Applicant: President and Fellows of Harvard College

Inventor： David A. Weitz ,  Huidan Zhang

IPC: C12Q1/6806 ,  C12Q1/6876 ,  C12Q1/686 ,  B01L3/00 ,  C12Q1/6834

Abstract: The present disclosure generally relates, in certain aspects, to droplet-based microfluidic devices and methods. In certain aspects, target nucleic acids contained within droplets are amplified within droplets in a first step, where multiple primers may be present. However, multiple primers may cause multiple target nucleic acids to be amplified within the droplets, which can make it difficult to identify which nucleic acids were amplified. In a second step, the amplified nucleic acids may be determined. For example, the droplets may be broken and the amplified nucleic acids can be pooled together and sequenced. As an example, new droplets may be formed containing the amplified nucleic acids, and those nucleic acids within the droplets amplified by exposure to certain primers.

公开(公告)号：US20230009208A1

公开(公告)日：2023-01-12

申请号：US17848523

申请日：2022-06-24

Applicant: The Brigham and Women’s Hospital, Inc. ,  President and Fellows of Harvard College ,  Vilnius University

Inventor： Joseph Italiano ,  Linas Mazutis ,  Jonathan N. Thon ,  David A. Weitz

IPC: B01L3/00 ,  C12M3/06 ,  C12M1/12 ,  C12M1/34 ,  G01N33/86 ,  C12M1/00

Abstract: 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.

公开(公告)号：US20220396835A1

公开(公告)日：2022-12-15

申请号：US17848909

申请日：2022-06-24

Applicant: President and Fellows of Harvard College

Inventor： David A. Weitz ,  Jeremy Agresti

IPC: C12Q1/6876 ,  B01J19/00 ,  C12Q1/6874 ,  B01J13/04

Abstract: The present invention is generally related to systems and methods for producing a plurality of droplets. The droplets may contain varying species, e.g., for use as a library. In some cases, the fluidic droplets may be rigidified to form rigidified droplets (e.g., gel droplets). In certain embodiments, the droplets may undergo a phase change (e.g., from rigidified droplets to fluidized droplets), as discussed more herein. In some cases, a species may be added internally to a droplet by exposing the droplet to a fluid comprising a plurality of species.

公开(公告)号：US20220274072A1

公开(公告)日：2022-09-01

申请号：US17745711

申请日：2022-05-16

Applicant: President and Fellows of Harvard College

Inventor： David A. Weitz ,  Adam R. Abate ,  Tony Hung ,  Pascaline Mary

IPC: B01F25/314 ,  F16K99/00 ,  B01J19/00 ,  B01L3/00 ,  B01F33/302 ,  B01F33/3031

Abstract: 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.

公开(公告)号：US20210379555A1

公开(公告)日：2021-12-09

申请号：US17323902

申请日：2021-05-18

Applicant: President and Fellows of Harvard College ,  Vilnius University

Inventor： David A. Weitz ,  Allon Moshe Klein ,  Ilke Akartuna ,  Linas Mazutis ,  Marc W. Kirschner

IPC: B01J19/00 ,  B01L3/00 ,  B01F13/00 ,  C12Q1/6806

Abstract: 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.

公开(公告)号：US20210348203A1

公开(公告)日：2021-11-11

申请号：US17148796

申请日：2021-01-14

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 M. Church

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

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.

公开(公告)号：US11001883B2

公开(公告)日：2021-05-11

申请号：US14478672

申请日：2014-09-05

Applicant: The General Hospital Corporation ,  President and Fellows of Harvard College

Inventor： Assaf Rotem ,  Oren Ram ,  Bradley E. Bernstein ,  David A. Weitz

IPC: C12Q1/68 ,  C07H21/00 ,  C12Q1/6869 ,  B01F13/00 ,  B01L3/00

Abstract: 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.

公开(公告)号：US10828641B2

公开(公告)日：2020-11-10

申请号：US15496750

申请日：2017-04-25

Applicant: President and Fellows of Harvard College

Inventor： Christian Boehm ,  Amy Rowat ,  Sarah Koester ,  Jeremy Agresti ,  David A. Weitz

IPC: B01L3/00 ,  B01L7/00 ,  G01N33/50 ,  G01N21/64

Abstract: 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.

公开(公告)号：US20200157593A1

公开(公告)日：2020-05-21

申请号：US16779501

申请日：2020-01-31

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 M. Church

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

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.