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公开(公告)号：US20090291433A1
公开(公告)日：2009-11-26
申请号：US11639490
申请日：2006-12-15
CPC分类号： C12Q1/68 , B01L7/52 , B01L2300/0645 , B01L2300/1822 , B01L2300/1894 , B01L2400/0424 , B01L2400/0427 , C12Q1/6869 , G01N27/44791 , Y10T436/25 , C12Q2565/629 , C12Q2565/301 , C12Q2563/149
摘要： The present invention relates to a droplet-based nucleic acid amplification method and apparatus. According to one embodiment, a method of amplifying a nucleic acid in a biological sample is provided, wherein the method includes: (a) providing a system comprising a droplet microactuator electronically coupled to and controlled by a processor capable of executing instructions, the droplet microactuator comprising: (i) a sample potentially comprising a target nucleic acid; (ii) a substrate comprising electrodes for conducting droplet operations; and (iii) one or more temperature control means arranged in proximity with one or more of the electrodes for heating a region of the droplet microactuator such that a droplet can be transported into the region for heating; (b) using droplet operations to combine on the droplet microactuator one or more amplification reagent droplets and one or more sample droplets to yield an amplification-ready droplet; and (c) thermal cycling the amplification-ready droplet sufficient to result in amplification of a target nucleic acid when present in the amplification-ready droplet.
摘要翻译： 本发明涉及一种基于液滴的核酸扩增方法和装置。 根据一个实施方案，提供了扩增生物样品中的核酸的方法，其中所述方法包括：（a）提供包括液滴微致动器的系统，所述液滴微致动器电子耦合到能够执行指令的处理器并由其控制，液滴微致动器 包括：（i）潜在地包含靶核酸的样品; （ii）包括用于进行液滴操作的电极的基板; 和（iii）设置在一个或多个电极附近的一个或多个温度控制装置，用于加热液滴微致动器的区域，使得液滴可以被运送到加热区域中; （b）使用液滴操作在液滴微致动器上组合一个或多个扩增试剂液滴和一个或多个样品液滴以产生扩增就绪液滴; 和（c）将放大准备好的液滴热循环到足以在存在于扩增即用液滴中时导致靶核酸的扩增。
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112.发明申请公开(公告)号：US20130233425A1
公开(公告)日：2013-09-12
申请号：US13866105
申请日：2013-04-19
IPC分类号： F16L55/00
CPC分类号： F16L55/00 , B01F13/0071 , B01F13/0076 , B01L3/502784 , B01L2200/143 , B01L2300/089 , B01L2300/161 , B01L2400/0427 , B01L2400/086 , Y10T137/206
摘要： The present invention is directed to modified droplet actuators, fluids and methods for enhancing and/or maintaining oil film stability in a droplet actuator. In an exemplary embodiment, the invention provides a droplet actuator with one or more substrates arranged to form a droplet operations gap comprising gap-facing surfaces; droplet operations electrodes configured to conduct droplet operations in the droplet operations gap; at least one barrier included on at least one of the substrate surfaces and having dimensions selected to: permit droplet transport from atop a first droplet operations electrode to a second droplet operations electrode when the second droplet operations electrode is activated; and prevent movement of a droplet from atop a first droplet operations electrode when the first and second droplet operations electrodes are inactive.
摘要翻译： 本发明涉及用于在液滴致动器中增强和/或维持油膜稳定性的改进的液滴致动器，流体和方法。 在示例性实施例中，本发明提供了具有一个或多个基板的液滴致动器，其被布置成形成包括间隙面的表面的液滴操作间隙; 液滴操作电极，被配置为在液滴操作间隙中进行液滴操作; 至少一个屏障包括在至少一个基板表面上，并且具有选定的尺寸：当第二液滴操作电极被激活时允许液滴从第一液滴操作电极顶部传输到第二液滴操作电极; 并且当第一和第二液滴操作电极不活动时，防止液滴从第一液滴操作电极的顶部移动。
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公开(公告)号：US08440392B2
公开(公告)日：2013-05-14
申请号：US12531844
申请日：2008-03-23
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 of conducting a droplet-based enzymatic assay is provided. The method generally makes use of a droplet actuator. A droplet comprising an enzyme of interest is provided on the droplet actuator along with a droplet comprising a substrate which is potentially modified in the presence of the enzyme. The method involves executing droplet operations on the droplet actuator to combine the droplets, thereby yielding an assay droplet, and detecting modification of the substrate by the enzyme in the assay droplet on the droplet actuator. The enzyme of interest may, for example, be a potentially mutated or improperly folded enzyme exhibiting altered enzyme activity as compared to a corresponding normal enzyme.
摘要翻译： 提供了一种进行基于液滴的酶测定的方法。 该方法通常使用液滴致动器。 包含感兴趣的酶的液滴与包含底物的液滴一起提供在液滴致动器上，所述液滴在酶的存在下可能被修饰。 该方法涉及对液滴致动器执行液滴操作以组合液滴，从而产生测定液滴，并且通过液滴致动器上的测定液滴中的酶检测底物的修饰。 感兴趣的酶可以是例如与相应的正常酶相比显示改变的酶活性的潜在突变或不正确折叠的酶。
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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：US20100120130A1
公开(公告)日：2010-05-13
申请号：US12647768
申请日：2009-12-28
CPC分类号： F16L55/00 , B01F13/0071 , B01F13/0076 , B01L3/502784 , B01L2200/143 , B01L2300/089 , B01L2300/161 , B01L2400/0427 , B01L2400/086 , Y10T137/206
摘要： The present invention is directed to droplet actuators with droplet retention structures, and methods related thereto. In an exemplary embodiment, the invention provides a droplet actuator with one or more substrates arranged to form a droplet operations gap comprising gap-facing surfaces; droplet operations electrodes configured to conduct droplet operations in the droplet operations gap; at least one barrier included on at least one of the substrate surfaces and having dimensions selected to: permit droplet transport from atop a first droplet operations electrode to a second droplet operations electrode when the second droplet operations electrode is activated; and prevent movement of a droplet from atop a first droplet operations electrode when the first and second droplet operations electrodes are inactive.
摘要翻译： 本发明涉及具有液滴保持结构的液滴致动器及其相关方法。 在示例性实施例中，本发明提供了具有一个或多个基板的液滴致动器，其被布置成形成包括间隙面的表面的液滴操作间隙; 液滴操作电极，被配置为在液滴操作间隙中进行液滴操作; 至少一个屏障包括在至少一个基板表面上，并且具有选定的尺寸：当第二液滴操作电极被激活时允许液滴从第一液滴操作电极顶部传输到第二液滴操作电极; 并且当第一和第二液滴操作电极不活动时，防止液滴从第一液滴操作电极的顶部移动。
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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：US20110104725A1
公开(公告)日：2011-05-05
申请号：US12990766
申请日：2009-05-04
申请人： Vamsee K. Pamula , Sitaram Emani , Allen Eckhardt , Carrie Graham , Jeremy Rouse
发明人： Vamsee K. Pamula , Sitaram Emani , Allen Eckhardt , Carrie Graham , Jeremy Rouse
IPC分类号： G01N33/53
CPC分类号： G01N33/86 , B01F13/0071 , B01F13/0076 , B01L3/502792 , B01L7/52 , B01L2200/0605 , B01L2200/0668 , B01L2300/0816 , B01L2300/0819 , B01L2300/089 , B01L2400/0406 , B01L2400/0409 , B01L2400/0424 , B01L2400/0427 , B01L2400/043 , B01L2400/046 , B01L2400/0487 , G01N33/5302
摘要： The invention provides techniques for coagulating blood on a droplet actuator. The invention also provides methods of manipulating the coagulated blood including a variety of droplet operations that may be conducted using the coagulated blood. Further, the invention provides a variety of assays that make use of the coagulated blood or various blood samples as input.
摘要翻译： 本发明提供了用于在液滴致动器上凝结血液的技术。 本发明还提供了操作凝血血液的方法，包括可以使用凝血血液进行的各种液滴操作。 此外，本发明提供使用凝血血液或各种血液样品作为输入的各种测定法。
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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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120.发明申请Enzymatic Assays Using Umbelliferone Substrates with Cyclodextrins in Droplets of Oil 有权使用伞形酮底物与环糊精在油滴中的酶检测公开(公告)号：US20110118132A1
公开(公告)日：2011-05-19
申请号：US13012831
申请日：2011-01-25
申请人： Theodore Winger , Allen E. Eckhardt , Carrie Graham , Ramakrishna Sista , Vamsee K. Pamula , Tong Wang , Yalin Xiong , Ning Wu , Sanjay Saha , Gajendrasinh Balvantsinh Raolji , Raveendra Dayam
发明人： Theodore Winger , Allen E. Eckhardt , Carrie Graham , Ramakrishna Sista , Vamsee K. Pamula , Tong Wang , Yalin Xiong , Ning Wu , Sanjay Saha , Gajendrasinh Balvantsinh Raolji , Raveendra Dayam
CPC分类号： C12Q1/00
摘要： The invention is directed to droplet actuator devices and assay methods. The method may include, among other things, a method of conducting enzymatic assays using umbelliferone substrates with cyclodextrins in droplets in oil, the method including incubating a droplet in oil, the droplet preferably comprising an umbelliferone substrate, a sample, and a cyclodextrin compound. The methods may further include a method of substantially eliminating cross-contamination between droplets in enzymatic containing substrate-based bioassays. The method may include immobilization of the enzymatic substrate including forming an inclusion complex with the substrate for stabilizing the substrate within an aqueous environment, wherein the inclusion complex may be formed using cyclodextrins. In yet another embodiment, the invention may provide a method of enhancing hydrolysis of enzymatic substrates. The methods may further include a method of forming an inclusion complex with the substrate for stabilizing the substrate within an aqueous environment, wherein the substrate may be a 4-MU- or HMU containing substrate. The inclusion complex may be formed using cyclodextrin.
摘要翻译： 本发明涉及液滴致动器装置和测定方法。 该方法尤其可以包括使用具有环糊精的伞形酮底物在油中的液滴进行酶测定的方法，所述方法包括将液滴在油中孵育，所述液滴优选包含伞形酮底物，样品和环糊精化合物。 所述方法还可以包括基本上消除含酶底物生物测定中的液滴之间的交叉污染的方法。 该方法可以包括固定酶底物，包括与底物形成包合物以使其在水性环境中稳定，其中包合物可以使用环糊精形成。 在另一个实施方案中，本发明可提供增强酶底物水解的方法。 所述方法还可以包括在水性环境中与基材形成包含配合物以稳定基材的方法，其中所述基材可以是含有4-MU-或HMU的底物。 包合物可以使用环糊精形成。
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