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    SYSTEMS AND METHODS FOR EPIGENETIC SEQUENCING
    1.
    发明公开
    SYSTEMS AND METHODS FOR EPIGENETIC SEQUENCING 审中-公开

    公开(公告)号：EP3305918A3

    公开(公告)日：2018-06-20

    申请号：EP17201280.9

    申请日：2013-03-05

    申请人： President and Fellows of Harvard College ,  The General Hospital Corporation d/b/a Massachusetts General Hospital

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

    IPC分类号： C12Q1/6869 ,  C12Q1/6806 ,  B01F13/00 ,  B01L3/00

    摘要： The present invention generally relates to microfluidics and 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.

    Method for forming multiple emulsions
    2.
    发明公开
    Method for forming multiple emulsions 有权
    Verfahren zur Bildung mehrerer Emulsionen

    公开(公告)号：EP2248578A1

    公开(公告)日：2010-11-10

    申请号：EP10165813.6

    申请日：2006-03-03

    申请人： President and Fellows of Harvard College

    发明人： Weitz, David A. ,  Link, Darren Roy ,  Utada, Andrew S.

    IPC分类号： B01F13/00 ,  B01F3/08 ,  B01F5/04 ,  B01J13/02

    CPC分类号： F15D1/008 ,  A61K9/113 ,  A61K9/1273 ,  A61K9/5089 ,  B01F3/0807 ,  B01F5/045 ,  B01F5/046 ,  B01F13/0059 ,  B01F13/0062 ,  B01F2005/0022 ,  B82Y10/00 ,  B82Y30/00 ,  Y10S516/924 ,  Y10T137/0391

    摘要： The present invention generally relates to a method for making multiple emulsions. A multiple emulsion generally describes larger droplets that contain one or more smaller droplets therein. The larger droplets may be suspended in a third fluid in some cases. These can be useful for encapsulating species such as pharmaceutical agents, cells, chemicals, or the like. In some cases, one or more of the droplets can change form, for instance, to become solidified to form a microcapsule, a liposome, a polymerosome, or a colloidosome, Multiple emulsions can be formed in one step in certain embodiments, with generally precise repeatability, and can be tailored to include one, two, three, or more inner droplets within a single outer droplet (which droplets may all be nested in some cases).

    摘要翻译： 本发明一般涉及制备多重乳液的方法。 多重乳液通常描述在其中含有一个或多个较小液滴的较大液滴。 在一些情况下，较大的液滴可以悬浮在第三流体中。 这些可用于包封物质如药剂，细胞，化学品等。 在一些情况下，一个或多个液滴可以改变形式，例如变得固化以形成微胶囊，脂质体，聚合物体或胶体体。在某些实施方案中，可以在一个步骤中形成多重乳液，其中通常精确 重复性，并且可以被定制为在单个外部液滴内包括一个，两个，三个或更多个内部液滴（在一些情况下，该液滴可以全部嵌套）。

    SYSTEMS AND METHODS FOR EPIGENETIC SEQUENCING
    3.
    发明公开
    SYSTEMS AND METHODS FOR EPIGENETIC SEQUENCING 审中-公开
    用于表观测序的系统和方法

    公开(公告)号：EP3305918A2

    公开(公告)日：2018-04-11

    申请号：EP17201280.9

    申请日：2013-03-05

    申请人： President and Fellows of Harvard College ,  The General Hospital Corporation d/b/a Massachusetts General Hospital

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

    IPC分类号： C12Q1/6869 ,  C12Q1/6806 ,  B01F13/00 ,  B01L3/00

    摘要： The present invention generally relates to microfluidics and 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.

    摘要翻译： 本发明一般涉及微流体和表观遗传测序。 在一组实施方案中，包含在多个微流体液滴内的细胞被裂解，并且液滴内的DNA（例如来自核小体）被标记，例如用含有识别序列的衔接子进行标记。 衔接子还可以包含其他序列，例如限制性位点，引物位点等，以辅助以后的分析。 在用衔接子标记后，来自不同细胞的DNA可以被组合并分析，例如以确定关于细胞的表观遗传信息。 例如，可以基于某些修饰（例如甲基化）分离DNA，并且可以使用诸如染色质免疫沉淀（“ChIP”）的技术对来自分离的核小体的DNA进行测序。 在一些情况下，DNA序列也可以与基因组进行比对， 例如，以确定基因组的哪些部分被表观遗传修饰， 例如通过甲基化。

    Method for mixing droplets in a microchannel
    4.
    发明公开
    Method for mixing droplets in a microchannel 审中-公开
    埃及米克罗卡纳尔（Vervahren Zur Mischung vonTröpfchen）

    公开(公告)号：EP2662135A2

    公开(公告)日：2013-11-13

    申请号：EP13165665.4

    申请日：2004-08-27

    申请人： President and Fellows of Harvard College

    发明人： Link, Darren R. ,  Weitz, David A. ,  Cristobal-Azkarte, Glader ,  Cheng, Zhengdong ,  Ahn, Keunho

    IPC分类号： B01J19/00 ,  G01N15/14

    CPC分类号： B01L3/5027 ,  B01F5/0256 ,  B01F5/0646 ,  B01F5/0655 ,  B01F5/0682 ,  B01F5/0689 ,  B01F13/0069 ,  B01F13/0071 ,  B01F13/0074 ,  B01F13/0076 ,  B01J19/0093 ,  B01J2219/00783 ,  B01J2219/00862 ,  B01L3/0241 ,  B01L3/502761 ,  B01L3/502776 ,  B01L3/502784 ,  B01L3/502792 ,  B01L3/5088 ,  B01L2200/0636 ,  B01L2200/0652 ,  B01L2200/0673 ,  B01L2300/0864 ,  B01L2300/0867 ,  B01L2300/161 ,  B01L2400/0415 ,  B01L2400/0439 ,  B01L2400/0487 ,  C12Q2563/159 ,  C12Q2565/629 ,  G01N15/10 ,  G01N15/14 ,  G01N15/1459 ,  G01N15/1484 ,  G01N2015/1006 ,  G01N2015/1081 ,  G01N2015/149 ,  Y10S436/807 ,  Y10T436/118339 ,  Y10T436/2525 ,  Y10T436/2575

    摘要： 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.

    摘要翻译： 本发明的各个方面涉及流体物质的控制和操作，例如在微流体系统中。 一方面，本发明涉及使用例如电场，机械改变，中间流体等的添加来制造由液体包围的流体液滴的系统和方法。在一些情况下，液滴可以具有 基本上统一的实体数量。 例如，95％或更多的液滴可以各自包含相同数量的特定物种的实体。 另一方面，本发明涉及用于将流体液滴分成两个液滴的系统和方法，例如通过与电场的电荷和/或偶极相互作用。 本发明还涉及根据本发明的另一方面，例如通过电荷和/或偶极相互作用来熔融液滴的系统和方法。 在一些情况下，液滴的融合可以引发或确定反应。 在本发明的相关方面，还提供了用于允许在液滴内进行流体混合的系统和方法。 在另一方面，本发明涉及用于分选液滴的系统和方法，例如通过使液滴移动到流体系统内的某些区域。 实例包括使用电相互作用（例如电荷，偶极子等）或机械系统（例如，流体位移）来对液滴进行分类。 在一些情况下，流体液滴可以以相对高的速率进行分选，例如每秒大约10个液滴或​​更多。 本发明的另一方面提供了例如使用荧光和/或其他光学技术（例如显微镜）或诸如电介质感测的电传感技术来确定液滴或其组分的能力。

    DROPLET CREATION TECHNIQUES
    8.
    发明公开
    DROPLET CREATION TECHNIQUES 审中-公开

    公开(公告)号：EP3461558A1

    公开(公告)日：2019-04-03

    申请号：EP18205385.0

    申请日：2010-10-26

    申请人： President and Fellows of Harvard College

    发明人： Weitz, David A. ,  Abate, Adam R.

    IPC分类号： B01L3/00 ,  B01F13/00

    摘要： 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.