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公开(公告)号：US08716015B2
公开(公告)日：2014-05-06
申请号：US12334575
申请日：2008-12-15
CPC分类号： C12N13/00 , B01J2219/00527 , B01J2219/00653 , B01J2219/00743 , G01N15/10 , G01N35/10 , G01N2015/0088 , G01N2015/1081 , G01N2035/00564 , G01N2035/1034 , Y10T436/11 , Y10T436/25 , Y10T436/2575
摘要： A method of inoculating a culture medium including providing a droplet including a single cell type on a droplet actuator and inoculating a culture medium with the droplet. A method of providing a metabolically useful substance to a cell culture, including providing a droplet actuator including a cell culture droplet loaded thereon, the sample droplet including cells and a cell culture medium, and a second droplet comprising a metabolically useful substance. The method also includes conducting one or more droplet operations to combine the cell culture droplet with the second droplet on the droplet actuator. Related methods, droplet actuators, and systems are also provided.
摘要翻译： 一种接种培养基的方法，包括在液滴致动器上提供包含单细胞类型的液滴并用液滴接种培养基。 提供代谢有用的物质给细胞培养物的方法，包括提供包括其上装载有细胞培养物液滴的液滴致动器，所述样品液滴包括细胞和细胞培养基，以及包含代谢有用物质的第二液滴。 该方法还包括进行一个或多个液滴操作以将细胞培养液滴与液滴致动器上的第二液滴组合。 还提供了相关方法，液滴致动器和系统。
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公开(公告)号：US20130230875A1
公开(公告)日：2013-09-05
申请号：US13862587
申请日：2013-04-15
IPC分类号： G01N21/64
CPC分类号： G01N21/6486 , B01F13/0809 , C07H15/04 , C12Q1/34 , G01N2333/928 , G01N2800/042
摘要： A method is provided for conducting a droplet-based enzymatic assay, e.g., for diagnostic purposes. On a droplet actuator, a droplet comprising an enzyme of interest is provided along with a droplet comprising a substrate which is potentially modified in the presence of the enzyme. Droplet operations are executed to combine the enzyme and substrate droplets on the droplet actuator, thereby yielding an assay droplet on the droplet actuator. Detecting modification of the substrate by the enzyme in the assay droplet occurs on the droplet actuator. Modified substrate preparations for conducting such enzymatic assays are also provided.
摘要翻译： 提供了用于进行基于液滴的酶测定的方法，例如用于诊断目的。 在液滴致动器上，包含感兴趣的酶的液滴与包含在酶存在下潜在地被修饰的底物的液滴一起提供。 执行液滴操作以将酶和底物液滴组合在液滴致动器上，从而在液滴致动器上产生测定液滴。 在液滴致动器中检测由测定液滴中的酶对底物的修饰。 还提供了用于进行这种酶测定的修饰底物制剂。
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公开(公告)号：US20100041086A1
公开(公告)日：2010-02-18
申请号：US12497677
申请日：2009-07-05
IPC分类号： C12Q1/34
CPC分类号： C12Q1/34 , B01F13/0071 , B01L3/502784 , B01L7/54 , B01L2300/0654 , B01L2300/0829 , B01L2300/0867 , B01L2300/089 , B01L2300/1827 , G01N2021/035 , G01N2333/924 , G01N2800/042
摘要： The invention relates to a microfluidic platform and methods of using the platform for conducting enzyme assays using a droplet actuator. The enzyme assays of the invention are useful for, among other things, identifying and/or characterizing disorders resulting from conditions in which enzymes are defective or are produced in inappropriate amounts. Enzyme assays of the invention may, for example, be used to detect altered activity of a particular enzyme in a sample, which may serve as an indicator of a particular disease. Altered activity may, for example, be caused by conditions which result in the increased or reduced production of a certain enzyme or its substrate and/or conditions which result in defective enzymes and/or substrates exhibiting increased or decreased effectiveness relative to corresponding normal enzymes and/or substrates.
摘要翻译： 本发明涉及一种微流体平台和使用该平台用于使用液滴致动器进行酶测定的方法。 本发明的酶测定法尤其可用于识别和/或表征由酶缺陷或以不适当量产生的病症引起的疾病。 本发明的酶测定可以例如用于检测样品中特定酶的改变的活性，其可以用作特定疾病的指标。 改变的活性可以例如由导致某些酶或其底物的产生增加或减少的条件引起，和/或导致有缺陷的酶和/或底物相对于相应的正常酶显示出增加或降低的有效性的条件，和 /或底物。
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公开(公告)号：US20090155902A1
公开(公告)日：2009-06-18
申请号：US12334575
申请日：2008-12-15
IPC分类号： C12N5/08
CPC分类号： C12N13/00 , B01J2219/00527 , B01J2219/00653 , B01J2219/00743 , G01N15/10 , G01N35/10 , G01N2015/0088 , G01N2015/1081 , G01N2035/00564 , G01N2035/1034 , Y10T436/11 , Y10T436/25 , Y10T436/2575
摘要： A method of inoculating a culture medium including providing a droplet including a single cell type on a droplet actuator and inoculating a culture medium with the droplet. A method of providing a metabolically useful substance to a cell culture, including providing a droplet actuator including a cell culture droplet loaded thereon, the sample droplet including cells and a cell culture medium, and a second droplet comprising a metabolically useful substance. The method also includes conducting one or more droplet operations to combine the cell culture droplet with the second droplet on the droplet actuator. Related methods, droplet actuators, and systems are also provided.
摘要翻译： 一种接种培养基的方法，包括在液滴致动器上提供包括单细胞类型的液滴并用液滴接种培养基。 提供代谢有用的物质给细胞培养物的方法，包括提供包括其上装载有细胞培养物液滴的液滴致动器，所述样品液滴包括细胞和细胞培养基，以及包含代谢有用物质的第二液滴。 该方法还包括进行一个或多个液滴操作以将细胞培养液滴与液滴致动器上的第二液滴组合。 还提供了相关方法，液滴致动器和系统。
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公开(公告)号：US08426213B2
公开(公告)日：2013-04-23
申请号：US12529052
申请日：2008-03-05
IPC分类号： G01N33/50
CPC分类号： C12Q1/28 , C12Q1/005 , C12Q1/006 , G01N2333/904 , Y10T436/118339 , Y10T436/14 , Y10T436/141111 , Y10T436/142222 , Y10T436/143333 , Y10T436/144444 , Y10T436/145555 , Y10T436/146666 , Y10T436/147777 , Y10T436/148888 , Y10T436/206664
摘要： The present invention relates to providing improved hydrogen peroxide assays, as well as droplet actuators for conducting such assays. The droplet actuators of the invention may be used to conduct droplet-based hydrogen peroxide assays. They may also be associated with detectors for analyzing the results of the hydrogen peroxide assays of the invention. They may be provided as components of systems which control droplet operations and/or detection for conducting the hydrogen peroxide assays. Measurement by the detector may be used to quantify the presence of an analyte in a sample.
摘要翻译： 本发明涉及提供改进的过氧化氢测定，以及用于进行这种测定的液滴致动器。 本发明的液滴致动器可用于进行基于液滴的过氧化氢测定。 它们也可以与用于分析本发明的过氧化氢测定结果的检测器相关联。 它们可以作为控制液滴操作和/或检测以进行过氧化氢测定的系统的组件提供。 检测器的测量可用于量化样品中分析物的存在。
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公开(公告)号：US08202686B2
公开(公告)日：2012-06-19
申请号：US12497677
申请日：2009-07-05
IPC分类号： C12Q1/00
CPC分类号： C12Q1/34 , B01F13/0071 , B01L3/502784 , B01L7/54 , B01L2300/0654 , B01L2300/0829 , B01L2300/0867 , B01L2300/089 , B01L2300/1827 , G01N2021/035 , G01N2333/924 , G01N2800/042
摘要： The invention relates to a microfluidic platform and methods of using the platform for conducting enzyme assays using a droplet actuator. The enzyme assays of the invention are useful for, among other things, identifying and/or characterizing disorders resulting from conditions in which enzymes are defective or are produced in inappropriate amounts. Enzyme assays of the invention may, for example, be used to detect altered activity of a particular enzyme in a sample, which may serve as an indicator of a particular disease. Altered activity may, for example, be caused by conditions which result in the increased or reduced production of a certain enzyme or its substrate and/or conditions which result in defective enzymes and/or substrates exhibiting increased or decreased effectiveness relative to corresponding normal enzymes and/or substrates.
摘要翻译： 本发明涉及一种微流体平台和使用该平台用于使用液滴致动器进行酶测定的方法。 本发明的酶测定法尤其可用于识别和/或表征由酶缺陷或以不适当量产生的病症引起的疾病。 本发明的酶测定可以例如用于检测样品中特定酶的改变的活性，其可以用作特定疾病的指标。 改变的活性可以例如由导致某些酶或其底物的产生增加或减少的条件引起，和/或导致有缺陷的酶和/或底物相对于相应的正常酶显示出增加或降低的有效性的条件，和 /或底物。
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公开(公告)号：US08828655B2
公开(公告)日：2014-09-09
申请号：US13862587
申请日：2013-04-15
IPC分类号： C12Q1/00
CPC分类号： G01N21/6486 , B01F13/0809 , C07H15/04 , C12Q1/34 , G01N2333/928 , G01N2800/042
摘要： A method is provided for conducting a droplet-based enzymatic assay, e.g., for diagnostic purposes. On a droplet actuator, a droplet comprising an enzyme of interest is provided along with a droplet comprising a substrate which is potentially modified in the presence of the enzyme. Droplet operations are executed to combine the enzyme and substrate droplets on the droplet actuator, thereby yielding an assay droplet on the droplet actuator. Detecting modification of the substrate by the enzyme in the assay droplet occurs on the droplet actuator. Modified substrate preparations for conducting such enzymatic assays are also provided.
摘要翻译： 提供了用于进行基于液滴的酶测定的方法，例如用于诊断目的。 在液滴致动器上，包含感兴趣的酶的液滴与包含在酶存在下潜在地被修饰的底物的液滴一起提供。 执行液滴操作以将酶和底物液滴组合在液滴致动器上，从而在液滴致动器上产生测定液滴。 在液滴致动器中检测由测定液滴中的酶对底物的修饰。 还提供了用于进行这种酶测定的修饰底物制剂。
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公开(公告)号：US20130288254A1
公开(公告)日：2013-10-31
申请号：US13390121
申请日：2010-08-13
申请人： Michael G. Pollack , Vijay Srinivasan , Allen E. Eckhardt , Prasanna Thwar , Zhishan Hua , Arjun Sudarsan , Theodore Winger
发明人： Michael G. Pollack , Vijay Srinivasan , Allen E. Eckhardt , Prasanna Thwar , Zhishan Hua , Arjun Sudarsan , Theodore Winger
IPC分类号： C12Q1/68
CPC分类号： G01N27/447 , B01L3/502761 , B01L3/502792 , B01L7/52 , B01L2200/0668 , B01L2300/0819 , B01L2300/0864 , B01L2300/0867 , B01L2300/089 , B01L2400/0427 , B01L2400/043 , C12Q1/686 , G01N33/54326 , G01N2035/00425 , G01N2035/1058 , G01N2035/1062 , C12Q2549/101 , C12Q2527/15 , C12Q2527/107 , C12Q2561/113
摘要： The invention is directed to certain droplet actuated molecular techniques. In one embodiment, the invention provides droplet actuator methods for detection of single nucleotide polymorphisms (SNPs) in a DNA sequence using digital microfluidics, including droplet actuator-based sample preparation and SNP analysis. In another embodiment, the invention provides droplet actuator devices and methods for providing integrated sample preparation and multiplexed detection of an infectious agent, such as HIV. In yet another embodiment, the invention provides droplet actuator devices and techniques for PCR amplification and detection of specific nucleic acid sequences using digital microfluidics, including droplet actuator-based sample preparation and target nucleic acid analysis. In yet another embodiment the invention provides methods for performing hot-start PCR on a droplet actuator. In yet another embodiment, the method of the invention combines PCR amplification with pyrosequencing to investigate specific sequences.
摘要翻译： 本发明涉及某些液滴致动分子技术。 在一个实施方案中，本发明提供用于使用数字微流体（包括基于液滴致动器的样品制备和SNP分析）检测DNA序列中的单核苷酸多态性（SNP）的液滴致动器方法。 在另一个实施方案中，本发明提供了液滴致动器装置和用于提供综合样品制备和多重检测感染因子如HIV的方法。 在另一个实施方案中，本发明提供了使用数字微流体（包括基于液滴致动器的样品制备和靶核酸分析）的PCR扩增和检测特异性核酸序列的液滴致动器装置和技术。 在另一个实施方案中，本发明提供了在液滴致动器上进行热启动PCR的方法。 在另一个实施方案中，本发明的方法将PCR扩增与焦磷酸测序结合以研究特异性序列。
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公开(公告)号：US20110311980A1
公开(公告)日：2011-12-22
申请号：US13139467
申请日：2009-12-15
CPC分类号： B01L3/502761 , B01L3/502784 , B01L3/502792 , B01L7/525 , B01L2200/10 , B01L2200/143 , B01L2300/0816 , B01L2300/0838 , B01L2400/0427 , B01L2400/043 , B01L2400/0644 , C12Q1/6869 , C12Q2565/301 , C12Q2565/626
摘要： The invention provides a droplet actuator device, as well as systems, methods and devices making use of the droplet actuator device. The droplet actuator device may include a substrate having electrodes arranged for conducting one or more droplet operations. The droplet actuator device may include a substrate having a reactor path with a wash region associated with a magnet for immobilizing mobilizing beads during bead washing operations. The droplet actuator device may include nucleotide base reservoirs and dedicated nucleotide base electrode paths arranged for transporting nucleotide base droplets from nucleotide base reservoirs to the reactor path. The droplet actuator device may include one or more wash buffer reservoirs associated with electrode paths arranged for transporting wash buffer droplets from wash buffer reservoirs to the reactor path. The droplet actuator device may include one or more sample reservoirs and sample paths arranged for transporting sample droplets from the one or more sample reservoirs to the reactor path. The droplet actuator device may include one or more enzyme reservoirs and dedicated enzyme electrode paths arranged for transporting enzyme droplets from the one or more enzyme reservoirs to a detection electrode.
摘要翻译： 本发明提供一种液滴致动器装置，以及利用液滴致动器装置的系统，方法和装置。 液滴致动器装置可以包括具有布置用于进行一个或多个液滴操作的电极的基板。 液滴致动器装置可以包括具有反应器路径的基底，其具有与磁珠相关联的洗涤区域，用于在珠洗涤操作期间固定动员珠粒。 液滴致动器装置可以包括核苷酸碱基储存器和专用核苷酸碱基电极路径，其设置用于将核苷酸碱基液滴从核苷酸碱基储存器输送到反应器路径。 液滴致动器装置可以包括一个或多个洗涤缓冲液储存器，其与设置用于将洗涤缓冲液滴从洗涤缓冲液储存器运送到反应器路径的电极路径相关联。 液滴致动器装置可以包括一个或多个样品储存器和样品路径，其被布置用于将样品液滴从一个或多个样品储存器输送到反应器路径。 液滴致动器装置可以包括一个或多个酶储器和用于将酶液滴从一个或多个酶储存器输送到检测电极的专用酶电极路径。
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公开(公告)号：US20100048410A1
公开(公告)日：2010-02-25
申请号：US12531826
申请日：2008-03-24
CPC分类号： G01N33/54366 , G01N33/54326 , G01N35/0098
摘要： A method of sorting beads on a droplet actuator. The method may, for example, include the following steps: (a) providing a droplet actuator comprising a substrate comprising electrodes arranged for conducting droplet operations on a substrate surface; (b) providing an assay droplet on the substrate surface, the droplet comprising two or more target-capture bead populations comprising target-capture beads comprising: (i) a capture probe bound to a target substance; and (ii) a unique bar binding element which binds to a corresponding binder; (c) using droplet operations to combine the assay droplet with a bead-capture droplet comprising one or more bead-capture beads having affinity for the binding element; (d) immobilizing the one or more bead-capture beads while conducting droplet operations to separate the bead-capture beads from unbound target-capture beads; (e) resuspending the one or more bead-capture beads in a droplet, thereby providing a droplet comprising a substantially pure substance-capture bead population; and (f) using droplet operations to conduct one or more protocol steps for an assay protocol.
摘要翻译： 一种在液滴致动器上分选珠粒的方法。 该方法可以例如包括以下步骤：（a）提供液滴致动器，其包括基板，该基板包括布置成用于在基板表面上进行液滴操作的电极; （b）在基底表面上提供测定液滴，所述液滴包含两个或更多个目标捕获珠群，包含目标捕获珠，其包含：（i）与靶物质结合的捕获探针; 和（ii）结合相应的粘合剂的独特的棒结合元件; （c）使用液滴操作将测定液滴与包含对结合元件具有亲和性的一个或多个珠粒捕获珠粒的珠粒捕获液滴组合; （d）固定所述一个或多个珠粒捕获珠粒，同时进行液滴操作以将珠粒捕获珠粒与未结合的靶捕获珠粒分离; （e）将一个或多个珠粒捕获珠重新悬浮在液滴中，由此提供包含基本上纯的物质捕获珠粒的液滴; 和（f）使用液滴操作为测定方案进行一个或多个方案步骤。
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