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1. 发明授权

US08809068B2 Manipulation of beads in droplets and methods for manipulating droplets 有权
标题翻译：操作小滴珠和操纵液滴的方法
公开(公告)号：US08809068B2
公开(公告)日：2014-08-19
申请号：US12761066
申请日：2010-04-15
申请人： Ramakrishna Sista , Vamsee K. Pamula , Vijay Srinivasan , Michael G. Pollack , Allen Eckhardt
发明人： Ramakrishna Sista , Vamsee K. Pamula , Vijay Srinivasan , Michael G. Pollack , Allen Eckhardt
IPC分类号： G01N15/06
CPC分类号： G01N33/54326 , B01F11/0071 , B01F13/0071 , B01F13/0076 , B01F13/0809 , B01L3/50273 , B01L3/502761 , B01L3/502792 , B01L7/52 , B01L7/525 , B01L2200/027 , B01L2200/0647 , B01L2200/0673 , B01L2300/06 , B01L2300/0654 , B01L2300/0816 , B01L2300/0861 , B01L2300/0864 , B01L2300/0867 , B01L2300/089 , B01L2300/1827 , B01L2400/0406 , B01L2400/0415 , B01L2400/0421 , B01L2400/0424 , B01L2400/0427 , B01L2400/043 , B01L2400/0436 , B01L2400/0442 , G01N1/38 , G01N2001/386 , G01N2035/1046 , Y10T436/11 , Y10T436/25 , Y10T436/25625 , Y10T436/2575
摘要： The invention provides a method of dispersing or circulating magnetically responsive beads within a droplet in a droplet actuator. The invention, in one embodiment, makes use of a droplet actuator with a plurality of droplet operations electrodes configured to transport the droplet, and a magnetic field present at a portion of the plurality of droplet operations electrodes. A bead-containing droplet is provided on the droplet actuator in the presence of the uniform magnetic field. Beads are circulated in the droplet during incubation by conducting droplet operations on the droplet within a uniform region of the magnetic field wherein droplet operations do not allow magnetically responsive beads to be introduced into a region of the magnetic field which is sufficiently non-uniform to cause bead clumping resulting in a more homogenous distribution of the beads in the droplet. Other embodiments are also provided.
摘要翻译：本发明提供了一种在液滴致动器中的液滴内分散或循环磁响应珠的方法。在一个实施例中，本发明利用具有配置成传送液滴的多个液滴操作电极的液滴致动器和存在于多个液滴操作电极的一部分处的磁场。在均匀磁场存在的情况下，在液滴致动器上提供含珠的液滴。在孵育期间，通过在磁场的均匀区域内对液滴进行液滴操作，珠在液滴中循环，其中液滴操作不允许将磁响应的珠引入磁场的区域，该区域足够不均匀，从而导致珠粒聚集，导致珠粒在液滴中更均匀的分布。还提供了其他实施例。

2. 发明申请

US20100279374A1 Manipulation of Beads in Droplets and Methods for Manipulating Droplets 有权
标题翻译：微滴控制珠粒和操纵水滴的方法
公开(公告)号：US20100279374A1
公开(公告)日：2010-11-04
申请号：US12761066
申请日：2010-04-15
申请人： Ramakrishna Sista , Vamsee K. Pamula , Vijay Srinivasan , Michael G. Pollack , Allen Eckhardt
发明人： Ramakrishna Sista , Vamsee K. Pamula , Vijay Srinivasan , Michael G. Pollack , Allen Eckhardt
IPC分类号： G01N1/10 , C12N13/00
CPC分类号： G01N33/54326 , B01F11/0071 , B01F13/0071 , B01F13/0076 , B01F13/0809 , B01L3/50273 , B01L3/502761 , B01L3/502792 , B01L7/52 , B01L7/525 , B01L2200/027 , B01L2200/0647 , B01L2200/0673 , B01L2300/06 , B01L2300/0654 , B01L2300/0816 , B01L2300/0861 , B01L2300/0864 , B01L2300/0867 , B01L2300/089 , B01L2300/1827 , B01L2400/0406 , B01L2400/0415 , B01L2400/0421 , B01L2400/0424 , B01L2400/0427 , B01L2400/043 , B01L2400/0436 , B01L2400/0442 , G01N1/38 , G01N2001/386 , G01N2035/1046 , Y10T436/11 , Y10T436/25 , Y10T436/25625 , Y10T436/2575
摘要： The invention provides a method of dispersing or circulating magnetically responsive beads within a droplet in a droplet actuator. The invention, in one embodiment, makes use of a droplet actuator with a plurality of droplet operations electrodes configured to transport the droplet, and a magnetic field present at a portion of the plurality of droplet operations electrodes. A bead-containing droplet is provided on the droplet actuator in the presence of the uniform magnetic field. Beads are circulated in the droplet during incubation by conducting droplet operations on the droplet within a uniform region of the magnetic field. Other embodiments are also provided.
摘要翻译：本发明提供了一种在液滴致动器中的液滴内分散或循环磁响应珠的方法。在一个实施例中，本发明利用具有配置成传送液滴的多个液滴操作电极的液滴致动器和存在于多个液滴操作电极的一部分处的磁场。在均匀磁场存在的情况下，在液滴致动器上提供含珠的液滴。在孵育期间，通过在磁场的均匀区域内的液滴上进行液滴操作，珠子在液滴中循环。还提供了其他实施例。

3. 发明申请

US20100068764A1 Droplet Actuator Devices and Methods Employing Magnetic Beads 有权
标题翻译：使用磁珠的液滴致动器装置和方法
公开(公告)号：US20100068764A1
公开(公告)日：2010-03-18
申请号：US12524495
申请日：2008-02-11
申请人： Ramakrishna Sista , Vamsee K. Pamula , Michael G. Pollack , Vijay Srinivasan , Allen E. Eckhardt
发明人： Ramakrishna Sista , Vamsee K. Pamula , Michael G. Pollack , Vijay Srinivasan , Allen E. Eckhardt
IPC分类号： G01N31/00 , C12P19/46
CPC分类号： G01N33/54326 , B01L3/502715 , B01L3/50273 , B01L3/502761 , B01L3/502784 , B01L2200/025 , B01L2200/028 , B01L2200/0647 , B01L2200/0673 , B01L2200/141 , B01L2300/0816 , B01L2300/0864 , B01L2300/0867 , B01L2400/0427 , B01L2400/043 , C12M25/01 , C12M25/16 , G01N27/44717 , G01N27/44786 , G01N33/54386 , G01N35/0098 , G01N2035/1046
摘要： A method of providing a droplet in contact with a magnetically responsive bead and having a reduced quantity of a substance. The method generally includes the steps of (a) providing a droplet actuator (200) comprising: (i) a substrate (210) comprising electrodes (214) arranged for conducting droplet operations on a surface; (ii) a starting droplet (218/222) comprising: (1) one or more magnetically responsive beads (220); (2) a starting quantity of the substance; and (3) a starting volume; (b) magnetically immobilizing the one or more magnetically responsive beads at a location which is at a distance from a target droplet splitting zone, (224); (c) conducting one or more droplet operations comprising droplet dividing operations selected to divide the combined droplet to yield a set of droplets comprising: (i) a droplet (218) comprising substantially all of the one or more magnetically responsive beads and having a decreased quantity of the substance relative to the starting concentration; and (ii) a droplet (222) substantially lacking the magnetically responsive beads.
摘要翻译：提供与磁响应珠接触并具有减少的物质量的液滴的方法。该方法通常包括以下步骤：（a）提供液滴致动器（200），包括：（i）包括布置用于在表面上进行液滴操作的电极（214）的基底（210）（ii）起始液滴（218/222），包括：（1）一个或多个磁响应珠（220）; （2）物质的起始量; 和（3）起始体积; （b）将所述一个或多个磁响应珠磁性地固定在距离目标液滴分离区一段距离的位置处（224）; （c）进行一个或多个液滴操作，其包括液滴分割操作，所述液滴分割操作被选择以分割组合的液滴以产生一组液滴，其包括：（i）液滴（218），其基本上包含所有一个或多个磁响应珠粒，物质的数量相对于起始浓度; 和（ii）基本上不含磁响应珠的液滴（222）。

4. 发明申请

US20100258441A1 Manipulation of Beads in Droplets and Methods for Splitting Droplets 有权
标题翻译：水滴中的珠子操作和分流水滴的方法
公开(公告)号：US20100258441A1
公开(公告)日：2010-10-14
申请号：US12761147
申请日：2010-04-15
申请人： Ramakrishna Sista , Vamsee K. Pamula , Vijay Srinivasan , Michael G. Pollack , Allen Eckhardt
发明人： Ramakrishna Sista , Vamsee K. Pamula , Vijay Srinivasan , Michael G. Pollack , Allen Eckhardt
IPC分类号： F04B19/00 , B81B7/00 , G01N27/26
CPC分类号： B01F11/0071 , B01F13/0071 , B01F13/0076 , B01L3/502746 , B01L3/502761 , B01L3/502792 , B01L7/52 , B01L7/525 , B01L2200/027 , B01L2300/0654 , B01L2300/0816 , B01L2300/0861 , B01L2300/0864 , B01L2300/0867 , B01L2300/089 , B01L2300/1827 , B01L2400/0406 , B01L2400/0415 , B01L2400/0421 , B01L2400/0424 , B01L2400/0427 , B01L2400/043 , B01L2400/0436 , B01L2400/0442 , G01N35/0098 , G01N2035/1046 , Y10T436/11 , Y10T436/25 , Y10T436/25625 , Y10T436/2575
摘要： The invention provides a method of circulating magnetically responsive beads within a droplet in a droplet actuator. The invention also provides methods for splitting droplets. The invention, in one embodiment, makes use of a droplet actuator with top and bottom substrates, a plurality of magnetic fields respectively present proximate the top and bottom substrates, wherein at least one of the magnet fields is selectively alterable, and a plurality of droplet operations electrodes positioned along at least one of the top and bottom surfaces. A droplet is positioned between the top and bottom surfaces and at least one of the magnetic fields is selectively altered.
摘要翻译：本发明提供了一种在液滴致动器中的液滴内循环磁响应珠的方法。本发明还提供了分裂液滴的方法。在一个实施例中，本发明利用具有顶部和底部基底的液滴致动器，分别存在于顶部和底部基底附近的多个磁场，其中至少一个磁场可选择性地改变，并且多个液滴沿着顶表面和底表面中的至少一个定位的操作电极。液滴位于顶表面和底表面之间，并且至少一个磁场被选择性地改变。

5. 发明申请

US20100236928A1 Multiplexed Detection Schemes for a Droplet Actuator 审中-公开
标题翻译：滴液致动器的多路检测方案
公开(公告)号：US20100236928A1
公开(公告)日：2010-09-23
申请号：US12681848
申请日：2008-10-15
申请人： Vijay Srinivasan , Vamsee K. Pamula , Ramakrishna Sista , Michael G. Pollack
发明人： Vijay Srinivasan , Vamsee K. Pamula , Ramakrishna Sista , Michael G. Pollack
IPC分类号： G01N35/10 , G01N27/26
CPC分类号： B01L3/502792 , B01L3/502761 , B01L2200/0673 , B01L2300/0654 , B01L2300/0816 , B01L2300/0819 , B01L2300/0861 , B01L2300/0864 , B01L2300/0867 , B01L2300/087 , B01L2300/088 , B01L2300/089 , B01L2400/0424 , B01L2400/0427 , B01L2400/0448
摘要： A droplet actualor having electrodes configured for ejecting droplet operations transporting droplets on a surface, and a sensor arranged in proximity to one or more of the electrodes establishing a detection window on the surface for detection of one or more properties of one or more droplets on the surface, wherein the electrodes establish at least two pathways for transport of droplets into the detection window. Also provided is, A method of detecting a property of a target droplet, including using droplet operations to modulate signals from a droplet set comprising the target droplet, detecting the modulated signals of the droplet set, demodulating the modulated signals to identify the signal produced by one or more individual droplets of the set Related methods and alternative embodiments are also provided.
摘要翻译：具有配置用于喷射在表面上输送液滴的液滴操作的电极的液滴实体，以及布置在一个或多个电极附近的传感器，其在表面上建立检测窗口，用于检测表面上的一个或多个液滴的一个或多个特性表面，其中电极建立用于将液滴输送到检测窗中的至少两个路径。还提供了一种检测目标液滴的属性的方法，包括使用液滴操作来调制来自包括目标液滴的液滴组的信号，检测液滴组的调制信号，解调调制信号以识别由还提供了一组或多个组合的相关方法和替代实施例。

6. 发明申请

US20110104747A1 Method of Concentrating Beads in a Droplet 有权
标题翻译：浓缩珠子在水滴中的方法
公开(公告)号：US20110104747A1
公开(公告)日：2011-05-05
申请号：US12921256
申请日：2009-03-09
申请人： Michael G. Pollack , Ramakrishna Sista , Zhishan Hua , Vamsee K. Pamula , Arjun Sudarsan , Vijay Srinivasan , Prasanna Thwar , Alexander Shenderov , Dwayne Allen
发明人： Michael G. Pollack , Ramakrishna Sista , Zhishan Hua , Vamsee K. Pamula , Arjun Sudarsan , Vijay Srinivasan , Prasanna Thwar , Alexander Shenderov , Dwayne Allen
IPC分类号： G01N1/40
CPC分类号： B01L3/0282 , B01F13/002 , B01F13/0023 , B01F13/0071 , B01F13/0076 , B01F13/0818 , B01L3/502753 , B01L2200/0621 , B01L2200/0631 , B01L2200/0652 , B01L2200/0668 , B01L2200/141 , B01L2300/0832 , B01L2300/0864 , B01L2300/0867 , B01L2400/043 , B03C1/01 , B03C1/284 , B03C1/286 , B03C2201/18 , B03C2201/26 , B03C2201/28 , Y10T436/25
摘要： Methods of concentrating beads in a droplet and/or loading beads on a fluidic device are provided, including among other things, a method of concentrating beads in a droplet, the method comprising: (a) providing a droplet actuator comprising: (i) an interior droplet operations volume; and (ii) a reservoir exterior to the interior volume; (iii) a droplet established in a liquid path extending from the reservoir into the interior volume; (b) providing magnetically responsive beads in the portion of the droplet which is in the reservoir; (c) magnetically attracting the magnetically responsive beads through the liquid path into the portion of the droplet which is in the interior volume; and (d) forming a droplet comprising one or more of the magnetically responsive beads in the interior volume.
摘要翻译：提供了将珠子集中在流体装置上的液滴和/或负载珠粒中的方法，其中包括将珠粒集中在液滴中的方法，所述方法包括：（a）提供液滴致动器，其包括：（i）内部液滴操作量; 和（ii）内部容积外部的储存器; （iii）建立在从储存器延伸到内部容积中的液体路径中的液滴; （b）在储存器中的液滴部分中提供磁响应珠; （c）通过所述液体路径将所述磁响应珠磁吸引到位于所述内部容积中的所述液滴的所述部分中; 和（d）在内部容积中形成包含一个或多个磁响应珠的液滴。

7. 发明申请

US20090263834A1 Droplet Actuator Devices and Methods for Immunoassays and Washing 有权
标题翻译：滴液致动器装置和免疫测定和洗涤方法
公开(公告)号：US20090263834A1
公开(公告)日：2009-10-22
申请号：US12393534
申请日：2009-02-26
申请人： Ramakrishna Sista , Michael G. Pollack , Vamsee K. Pamula , Vijay Srinivasan
发明人： Ramakrishna Sista , Michael G. Pollack , Vamsee K. Pamula , Vijay Srinivasan
IPC分类号： G01N33/542
CPC分类号： G01N33/54333
摘要： Droplet actuator devices and methods for immunoassays and washing are provided. According to one embodiment, a method of providing a droplet in contact with a surface of a super paramagnetic bead with a reduced concentration of a substance is provided and includes: (a) providing a super paramagnetic bead in contact with a droplet comprising a starting concentration and starting quantity of the substance and having a starting volume; (b) conducting one or more droplet operations to merge a wash droplet with the droplet provided in step (a) to yield a combined droplet; and (c) conducting one or more droplet operations to divide the combined droplet to yield a set of droplets. The set of droplets includes: (i) a droplet in contact with the super paramagnetic bead having a decreased concentration of the substance relative to the starting concentration; and (ii) a droplet which is separated from the super paramagnetic bead.
摘要翻译：提供液滴致动器装置和免疫测定和洗涤方法。根据一个实施方案，提供了一种提供与物质浓度降低的超顺磁珠的表面接触的液滴的方法，包括：（a）提供与包含起始浓度的液滴接触的超顺磁珠并开始物质的起始体积; （b）进行一个或多个液滴操作以将洗涤液滴与步骤（a）中提供的液滴合并以产生组合液滴; 和（c）进行一个或多个液滴操作以分开组合液滴以产生一组液滴。一组液滴包括：（i）与超顺磁珠接触的液滴，其物质相对于起始浓度降低; 和（ii）从超顺磁珠分离的液滴。

8. 发明授权

US09675972B2 Method of concentrating beads in a droplet 有权
公开(公告)号：US09675972B2
公开(公告)日：2017-06-13
申请号：US12921256
申请日：2009-03-09
申请人： Michael G. Pollack , Ramakrishna Sista , Zhishan Hua , Vamsee K. Pamula , Arjun Sudarsan , Vijay Srinivasan , Prasanna Thwar , Alexander Shenderov , Dwayne Allen
发明人： Michael G. Pollack , Ramakrishna Sista , Zhishan Hua , Vamsee K. Pamula , Arjun Sudarsan , Vijay Srinivasan , Prasanna Thwar , Alexander Shenderov , Dwayne Allen
IPC分类号： B01L3/00 , B01L3/02 , B01F13/00 , B01F13/08 , B03C1/01 , B03C1/28
CPC分类号： B01L3/0282 , B01F13/002 , B01F13/0023 , B01F13/0071 , B01F13/0076 , B01F13/0818 , B01L3/502753 , B01L2200/0621 , B01L2200/0631 , B01L2200/0652 , B01L2200/0668 , B01L2200/141 , B01L2300/0832 , B01L2300/0864 , B01L2300/0867 , B01L2400/043 , B03C1/01 , B03C1/284 , B03C1/286 , B03C2201/18 , B03C2201/26 , B03C2201/28 , Y10T436/25
摘要： Methods of concentrating beads in a droplet and/or loading beads on a fluidic device are provided, including among other things, a method of concentrating beads in a droplet, the method comprising: (a) providing a droplet actuator comprising: (i) an interior droplet operations volume; and (ii) a reservoir exterior to the interior volume; (iii) a droplet established in a liquid path extending from the reservoir into the interior volume; (b) providing magnetically responsive beads in the portion of the droplet which is in the reservoir; (c) magnetically attracting the magnetically responsive beads through the liquid path into the portion of the droplet which is in the interior volume; and (d) forming a droplet comprising one or more of the magnetically responsive beads in the interior volume.

9. 发明授权

US09046514B2 Droplet actuator devices and methods employing magnetic beads 有权
标题翻译：液滴致动器装置和采用磁珠的方法
公开(公告)号：US09046514B2
公开(公告)日：2015-06-02
申请号：US12524495
申请日：2008-02-11
申请人： Ramakrishna Sista , Vamsee K. Pamula , Michael G. Pollack , Vijay Srinivasan , Allen E. Eckhardt , Richard B. Fair
发明人： Ramakrishna Sista , Vamsee K. Pamula , Michael G. Pollack , Vijay Srinivasan , Allen E. Eckhardt , Richard B. Fair
IPC分类号： G01N31/00 , G01N33/543 , B01L3/00 , C12M1/12 , C12P19/46 , G01N35/10 , G01N35/00
CPC分类号： G01N33/54326 , B01L3/502715 , B01L3/50273 , B01L3/502761 , B01L3/502784 , B01L2200/025 , B01L2200/028 , B01L2200/0647 , B01L2200/0673 , B01L2200/141 , B01L2300/0816 , B01L2300/0864 , B01L2300/0867 , B01L2400/0427 , B01L2400/043 , C12M25/01 , C12M25/16 , G01N27/44717 , G01N27/44786 , G01N33/54386 , G01N35/0098 , G01N2035/1046
摘要： A method of providing a droplet in contact with a magnetically responsive bead and having a reduced quantity of a substance. The method generally includes the steps of (a) providing a droplet actuator (200) comprising: (i) a substrate (210) comprising electrodes (214) arranged for conducting droplet operations on a surface; (ii) a starting droplet (218/222) comprising: (1) one or more magnetically responsive beads (220); (2) a starting quantity of the substance; and (3) a starting volume; (b) magnetically immobilizing the one or more magnetically responsive beads at a location which is at a distance from a target droplet splitting zone, (224); (c) conducting one or more droplet operations comprising droplet dividing operations selected to divide the combined droplet to yield a set of droplets comprising: (i) a droplet (218) comprising substantially all of the one or more magnetically responsive beads and having a decreased quantity of the substance relative to the starting concentration; and (ii) a droplet (222) substantially lacking the magnetically responsive beads.
摘要翻译：提供与磁响应珠接触并具有减少的物质量的液滴的方法。该方法通常包括以下步骤：（a）提供液滴致动器（200），包括：（i）包括布置用于在表面上进行液滴操作的电极（214）的基底（210）（ii）起始液滴（218/222），包括：（1）一个或多个磁响应珠（220）; （2）物质的起始量; 和（3）起始体积; （b）将所述一个或多个磁响应珠磁性地固定在距离目标液滴分离区一段距离的位置处（224）; （c）进行一个或多个液滴操作，其包括液滴分割操作，所述液滴分割操作被选择以分割组合的液滴以产生一组液滴，其包括：（i）液滴（218），其基本上包含所有一个或多个磁响应珠粒，物质的数量相对于起始浓度; 和（ii）基本上不含磁响应珠的液滴（222）。

10. 发明授权

US08846414B2 Detection of cardiac markers on a droplet actuator 有权
标题翻译：液滴致动器上心脏标志物的检测
公开(公告)号：US08846414B2
公开(公告)日：2014-09-30
申请号：US12569725
申请日：2009-09-29
申请人： Ramakrishna Sista , Vamsee K. Pamula , Arjun Sudarsan , Vijay Srinivasan , Michael G. Pollack , Richard B. Fair , Allen E. Eckhardt
发明人： Ramakrishna Sista , Vamsee K. Pamula , Arjun Sudarsan , Vijay Srinivasan , Michael G. Pollack , Richard B. Fair , Allen E. Eckhardt
IPC分类号： G01N33/553 , G01N33/68 , G01N33/543
CPC分类号： G01N33/6893 , G01N33/54366 , G01N2333/4712 , G01N2333/58 , G01N2800/32 , Y10T436/25
摘要： The present invention provides for the detection of cardiac markers on a droplet actuator. An aspect provides a method of assaying a cardiac marker in a biological sample from a subject, the method including providing a droplet actuator, loading the biological sample and assay reagents on the droplet actuator, executing droplet operations to create sample droplets from the sample and reagent droplets from the reagents on the droplet actuator, and executing droplet operations using the sample droplets and reagent droplets to produce a detectable signal indicative of the quantity of the cardiac marker in the biological sample. Still other aspects are provided.
摘要翻译：本发明提供了在液滴致动器上检测心脏标志物。一方面提供了从受试者的生物样品中测定心脏标志物的方法，所述方法包括提供液滴致动器，在液滴致动器上装载生物样品和测定试剂，执行液滴操作以从样品和试剂产生样品液滴来自液滴致动器上的试剂的液滴，以及使用样品液滴和试剂液滴执行液滴操作，以产生指示生物样品中心脏标志物量的可检测信号。还提供了其他方面。

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