公开(公告)号：US08852952B2

公开(公告)日：2014-10-07

申请号：US12990815

申请日：2009-05-04

Applicant: Michael G. Pollack ,  Prasanna Thwar ,  Vamsee K. Pamula ,  Allen Eckhardt ,  Alexander Shenderov ,  Dwayne Allen ,  Vijay Srinivasan

Inventor： Michael G. Pollack ,  Prasanna Thwar ,  Vamsee K. Pamula ,  Allen Eckhardt ,  Alexander Shenderov ,  Dwayne Allen ,  Vijay Srinivasan

IPC: G01N1/00

CPC classification number: B01L3/502784 ,  B01L3/502715 ,  B01L3/502761 ,  B01L7/52 ,  B01L2200/027 ,  B01L2200/0673 ,  B01L2300/022 ,  B01L2300/023 ,  B01L2300/024 ,  B01L2300/0672 ,  B01L2300/0816 ,  B01L2300/0864 ,  B01L2300/0867 ,  B01L2400/0415 ,  B01L2400/0424 ,  B01L2400/0427 ,  B01L2400/043 ,  B01L2400/0478 ,  B01L2400/0481 ,  B01L2400/0487 ,  B01L2400/049 ,  B01L2400/0683 ,  G01N27/44704 ,  G01N35/0098 ,  G01N2035/1046 ,  Y10T436/25 ,  Y10T436/2575

Abstract: The invention provides droplet actuators and droplet actuator cassettes including reagent storage capabilities, as well as methods of making and using the droplet actuators and cassettes. The invention also provides continuous flow channel elements and techniques for using electrodes to manipulate droplets in flowing streams. The invention also discloses methods of separating compounds on a droplet actuator. Various other aspects of the invention are also disclosed.

Abstract translation: 本发明提供了包括试剂储存能力的液滴致动器和液滴致动器盒，以及制造和使用液滴致动器和盒的方法。 本发明还提供了用于使用电极在流动流中操纵液滴的连续流动通道元件和技术。 本发明还公开了在液滴致动器上分离化合物的方法。 还公开了本发明的各种其它方面。

公开(公告)号：US08809068B2

公开(公告)日：2014-08-19

申请号：US12761066

申请日：2010-04-15

Applicant: Ramakrishna Sista ,  Vamsee K. Pamula ,  Vijay Srinivasan ,  Michael G. Pollack ,  Allen Eckhardt

Inventor： Ramakrishna Sista ,  Vamsee K. Pamula ,  Vijay Srinivasan ,  Michael G. Pollack ,  Allen Eckhardt

IPC: G01N15/06

CPC classification number: 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

Abstract: 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.

Abstract translation: 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。

公开(公告)号：US20140193807A1

公开(公告)日：2014-07-10

申请号：US14163442

申请日：2014-01-24

Applicant: ADVANCED LIQUID LOGIC, INC.

Inventor： VAMSEE K. PAMULA ,  ARJUN SUDARSAN ,  RAMAKRISHNA SISTA

IPC: C12Q1/68 ,  G01N33/543

CPC classification number: B03C7/026 ,  B01L3/50273 ,  B01L3/502761 ,  B01L2200/0647 ,  B01L2300/0887 ,  B01L2400/0424 ,  B01L2400/0427 ,  B03C1/01 ,  B03C1/288 ,  B03C5/00 ,  B03C5/005 ,  B03C5/026 ,  B03C2201/26 ,  G01N33/54326

Abstract: The invention provides a method of redistributing magnetically responsive beads in a droplet. The method may include providing a droplet including magnetically responsive beads. The droplet may be provided within a region of a magnetic field having sufficient strength to attract the magnetically responsive beads to an edge of the droplet or towards an edge of the droplet, or otherwise regionalize or aggregate beads within the droplet. The method may also include conducting on a droplet operations surface one or more droplet operations using the droplet without removing the magnetically responsive beads from the region of the magnetic field. The droplet operations may in some cases be electrode-mediated. The droplet operations may redistribute and/or circulate the magnetically responsive beads within the droplet. In some cases, the droplet may include a sample droplet may include a target analyte. The redistributing of the magnetically responsive beads may cause target analyte to bind to the magnetically responsive beads. In some cases, the droplet may include unbound substances in a wash buffer. The redistributing of the magnetically responsive beads causes unbound substances to be freed from interstices of an aggregated set or subset of the magnetically responsive beads.

Abstract translation: 本发明提供了一种在液滴中重新分布磁响应珠的方法。 该方法可以包括提供包括磁响应珠的液滴。 液滴可以设置在具有足够强度的磁场的区域内，以将磁响应珠吸引到液滴的边缘或朝向液滴的边缘，或以其他方式区分或聚集液滴内的珠粒。 该方法还可以包括在液滴操作表面上使用液滴进行一个或多个液滴操作，而不从磁场的区域移除磁响应珠。 在某些情况下，液滴操作可能是电极介导的。 液滴操作可以在液滴内重新分布和/或循环磁响应珠。 在一些情况下，液滴可以包括样品液滴可以包括靶分析物。 磁响应珠的重新分布可能导致目标分析物结合磁响应珠。 在一些情况下，液滴可以包括洗涤缓冲液中的未结合物质。 磁响应珠子的重新分布导致未结合物质从磁性响应珠粒的聚集或子集的空隙中释放出来。

公开(公告)号：US20140161686A1

公开(公告)日：2014-06-12

申请号：US14102367

申请日：2013-12-10

Applicant: Advanced Liquid Logic, Inc.

Inventor： Donovan E. BORT ,  Vijay SRINIVASAN ,  Patrick STERLINA ,  Uichong B. YI ,  Justin CAPPELLETTI

IPC: B01L3/00

CPC classification number: B01L3/527 ,  B01L3/502715 ,  B01L3/502784 ,  B01L7/52 ,  B01L2200/027 ,  B01L2200/0673 ,  B01L2200/16 ,  B01L2300/045 ,  B01L2300/049 ,  B01L2300/0672 ,  B01L2300/0816 ,  B01L2300/0867 ,  B01L2300/1827 ,  B01L2400/0427 ,  B01L2400/0644 ,  B01L2400/065 ,  B01L2400/0683

Abstract: Microfluidic system including a droplet actuator having an interior cavity and a series of electrodes arranged along the interior cavity for forming a droplet-operation path therethrough. The droplet actuator has a module-engaging side including an opening that is in flow communication with the interior cavity. The microfluidic system also includes a reservoir module configured to be coupled to the droplet actuator. The reservoir module includes a plurality of liquid compartments having respective outlets and at least one seal positioned along the outlets to retain liquid within the liquid compartments. The reservoir module is movable along the module-engaging side of the droplet actuator to position the outlets relative to the opening. The microfluidic system also includes a piercer having a tip configured to penetrate the seal thereby permitting the liquid within the corresponding liquid compartment to flow into the opening.

Abstract translation: 微流体系统包括具有内腔的液滴致动器和沿内腔布置的一系列电极，用于形成通过其的液滴操作路径。 液滴致动器具有包括与内腔流动连通的开口的模块接合侧。 微流体系统还包括被配置为耦合到液滴致动器的贮存器模块。 储存器模块包括多个具有相应出口的液体隔室和沿出口定位的至少一个密封件，以将液体保持在液体隔间内。 储存器模块可沿液滴致动器的模块接合侧移动，以使出口相对于开口定位。 微流体系统还包括具有尖端的穿孔器，该尖端被配置为穿透密封，从而允许相应的液体隔室内的液体流入开口。

公开(公告)号：US08734629B2

公开(公告)日：2014-05-27

申请号：US13101787

申请日：2011-05-05

Applicant: Alexander Shenderov

Inventor： Alexander Shenderov

IPC: G01N27/447 ,  G01N27/453

CPC classification number: B01L3/502792 ,  B01F13/0071 ,  B01F13/0076 ,  B01F2005/0017 ,  B01L3/502738 ,  B01L3/502746 ,  B01L3/502784 ,  B01L2200/0673 ,  B01L2200/10 ,  B01L2200/16 ,  B01L2300/0816 ,  B01L2300/0819 ,  B01L2300/0867 ,  B01L2300/089 ,  B01L2400/0415 ,  B01L2400/0427 ,  F04B17/00 ,  F04B19/006 ,  F16K99/0001 ,  F16K99/0017 ,  G02B26/005

Abstract: A series of microactuators for manipulating small quantities of liquids, and methods of using these for manipulating liquids, are disclosed. The microactuators are based on the phenomenon of electrowetting and contain no moving parts. The force acting on the liquid is a potential-dependent gradient of adhesion energy between the liquid and a solid insulating surface.

Abstract translation: 公开了一系列用于操作少量液体的微致动器以及使用它们来操纵液体的方法。 微致动器基于电润湿现象，不含运动部件。 作用在液体上的力是液体和固体绝缘表面之间的粘附能的电势梯度。

公开(公告)号：US20140141460A1

公开(公告)日：2014-05-22

申请号：US14089070

申请日：2013-11-25

Applicant: Advanced Liquid Logic, Inc.

Inventor： Allen E. Eckhardt

IPC: C12Q1/54

CPC classification number: C12Q1/40 ,  B01L3/502784 ,  B01L2300/0654 ,  B01L2300/0816 ,  B01L2300/18 ,  B01L2400/0427 ,  B01L2400/043 ,  C07D311/16 ,  C07D311/18 ,  C07D311/20 ,  C07D405/12 ,  C07D413/12 ,  C07H15/203 ,  C07H17/075 ,  C12Q1/25 ,  C12Q1/34 ,  C12Q1/44 ,  C12Q1/54 ,  C12Y302/01023 ,  G01N21/6452 ,  G01N33/5438 ,  G01N2333/916 ,  G01N2333/926 ,  G01N2333/938 ,  G01N2800/04

Abstract: The invention is directed to droplet actuator devices and assay methods. The invention includes assay methods of conducting an assay comprising combining a sample with an umbelliferyl derivative, wherein the sample potentially comprises an enzyme capable of cleaving the umbelliferyl derivative and where the umbelliferyl derivative comprises an umbelliferyl core modified with one or more modifying moieties.

Abstract translation: 本发明涉及液滴致动器装置和测定方法。 本发明包括进行测定的测定方法，其包括将样品与伞形酮基衍生物组合，其中所述样品潜在地包含能够切割伞形变体基衍生物的酶，并且其中伞形载体衍生物包含用一个或多个修饰部分修饰的伞形酮核心。

公开(公告)号：US20130293246A1

公开(公告)日：2013-11-07

申请号：US13988190

申请日：2011-11-15

Applicant: Michael G. Pollack ,  Ryan A. Sturmer ,  Philip Y. Paik ,  Vamsee K. Pamula ,  Vijay Srinivasan

Inventor： Michael G. Pollack ,  Ryan A. Sturmer ,  Philip Y. Paik ,  Vamsee K. Pamula ,  Vijay Srinivasan

IPC: G01N27/22 ,  G01K7/34

CPC classification number: G01N27/221 ,  B01L3/502784 ,  B01L3/502792 ,  B01L2200/14 ,  B01L2300/0645 ,  B01L2300/0816 ,  B01L2300/089 ,  B01L2400/0427 ,  B41J2/04541 ,  B41J2/04555 ,  B41J2/0458 ,  G01K7/34

Abstract: The embodiments described herein provide methods of measuring capacitance, detecting a droplet at a position, determining a thickness of an oil film and determining temperature in a droplet actuator. Specifically, the capacitance detection method may be used as a real-time verification tool in order to detect the absence, presence, and/or partial presence of a droplet at an electrode, analyze droplet properties, measure droplet size or volume, optimize the speed of droplet operation and detect air bubbles.

Abstract translation: 本文所述的实施例提供了测量电容，检测位置处的液滴，确定油膜的厚度和确定液滴致动器中的温度的方法。 具体地，可以将电容检测方法用作实时验证工具，以便检测电极处的液滴的不存在，存在和/或部分存在，分析液滴属性，测量液滴尺寸或体积，优化速度 的液滴操作并检测气泡。

公开(公告)号：US20130230875A1

公开(公告)日：2013-09-05

申请号：US13862587

申请日：2013-04-15

Applicant: Vamsee K. Pamula ,  Allen E. Eckhardt ,  Jeremy Rouse ,  Vijay Srinivasan

Inventor： Vamsee K. Pamula ,  Allen E. Eckhardt ,  Jeremy Rouse ,  Vijay Srinivasan

IPC: G01N21/64

CPC classification number: G01N21/6486 ,  B01F13/0809 ,  C07H15/04 ,  C12Q1/34 ,  G01N2333/928 ,  G01N2800/042

Abstract: 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.

Abstract translation: 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。

公开(公告)号：US20130225452A1

公开(公告)日：2013-08-29

申请号：US13596965

申请日：2012-08-28

Applicant: Michael G. Pollack ,  Allen E. Eckhardt ,  Prasanna Thwar ,  Jeremy Rouse

Inventor： Michael G. Pollack ,  Allen E. Eckhardt ,  Prasanna Thwar ,  Jeremy Rouse

IPC: C12N15/10

CPC classification number: C12N15/1075 ,  B01J19/0046 ,  C12N15/1006 ,  C40B40/08 ,  C40B50/06

Abstract: A method of preparing a nucleic acid library in droplets in contact with oil, including: (a) blunt-ending nucleic acid fragments in a droplet in the oil to yield blunt-ended nucleic acid fragments;(b) phosphorylating the blunt-ended nucleic acid fragments in a droplet in the oil to yield phosphorylated nucleic acid fragments; coupling A-tails to the phosphorylated nucleic acid fragments in a droplet in the oil to yield A-tailed nucleic acid fragments; and (d) coupling nucleic acid adapters to the A-tailed nucleic acid fragments in a droplet in the oil to yield the nucleic acid library comprising adapter-ligated nucleic acid fragments.

公开(公告)号：US20130225450A1

公开(公告)日：2013-08-29

申请号：US13596548

申请日：2012-08-28

Applicant: Michael G. Pollack ,  Allen E. Eckhardt ,  Prasanna Thwar ,  Jeremy Rouse

Inventor： Michael G. Pollack ,  Allen E. Eckhardt ,  Prasanna Thwar ,  Jeremy Rouse

IPC: B01J19/00

CPC classification number: C12N15/1075 ,  B01J19/0046 ,  C12N15/1006 ,  C40B40/08 ,  C40B50/06

Abstract: A method of preparing a nucleic acid library in droplets in contact with oil, including: (a) blunt-ending nucleic acid fragments in a droplet in the oil to yield blunt-ended nucleic acid fragments; (b) phosphorylating the blunt-ended nucleic acid fragments in a droplet in the oil to yield phosphorylated nucleic acid fragments; coupling A-tails to the phosphorylated nucleic acid fragments in a droplet in the oil to yield A-tailed nucleic acid fragments; and (d) coupling nucleic acid adapters to the A-tailed nucleic acid fragments in a droplet in the oil to yield the nucleic acid library comprising adapter-ligated nucleic acid fragments.

Abstract translation: 制备与油接触的液滴核酸文库的方法，包括：（a）在油中的液滴中的平端核酸片段以产生平端核酸片段; （b）使油中的液滴中的平端核酸片段磷酸化以产生磷酸化的核酸片段; 将A尾连接到油中的液滴中的磷酸化核酸片段以产生A尾核酸片段; 和（d）将核酸衔接子偶联到油中的液滴中的A尾核酸片段，以产生包含衔接子连接的核酸片段的核酸文库。