公开(公告)号：US09383295B2

公开(公告)日：2016-07-05

申请号：US14477642

申请日：2014-09-04

Applicant: Fluidigm Corporation

Inventor： Martin Pieprzyk ,  Geoff Facer ,  Timothy Woudenberg ,  Brian Fowler

IPC: G01N21/64 ,  G01N1/28 ,  B01L3/00 ,  G01N21/05 ,  C12Q1/68 ,  B01L7/00 ,  G01N21/03

CPC classification number: G01N1/28 ,  B01L3/502738 ,  B01L7/52 ,  B01L2200/16 ,  B01L2300/0867 ,  B01L2300/0874 ,  B01L2300/123 ,  B01L2400/0481 ,  B01L2400/0655 ,  B33Y80/00 ,  C12Q1/686 ,  G01N21/05 ,  G01N21/6452 ,  G01N2021/0346 ,  Y10T137/7793 ,  Y10T436/2575

Abstract: Embodiments of the present invention provide improved microfluidic devices and related apparatus, systems, and methods. Methods are provided for reducing mixing times during use of microfluidic devices. Microfluidic devices and related methods of manufacturing are provided with increased manufacturing yield rates. Improved apparatus and related systems are provided for supplying controlled pressure to microfluidic devices. Methods and related microfluidic devices are provided for reducing dehydration of microfluidic devices during use. Microfluidic devices and related methods are provided with improved sample to reagent mixture ratio control. Microfluidic devices and systems are provided with improved resistance to compression fixture pressure induced failures. Methods and systems for conducting temperature controlled reactions using microfluidic devices are provided that reduce condensation levels within the microfluidic device. Methods and systems are provided for improved fluorescent imaging of microfluidic devices.

公开(公告)号：US09249459B2

公开(公告)日：2016-02-02

申请号：US14102331

申请日：2013-12-10

Applicant: Fluidigm Corporation

Inventor： Amy Hamilton ,  Min Lin ,  Alain Mir ,  Martin Pieprzyk

IPC: C12Q1/68 ,  C12P19/34

CPC classification number: C12Q1/6881 ,  C12Q1/6844 ,  C12Q1/686 ,  C12Q2531/113 ,  C12Q2531/119 ,  C12Q2531/137 ,  C12Q2565/101 ,  C12Q2600/156 ,  C12Q2600/158 ,  C12Q2600/16 ,  C12Q2600/178

Abstract: The present invention provides methods for analysis of genomic DNA and/or RNA from small samples or even single cells. Methods for analyzing genomic DNA can entail whole genome amplification (WGA), followed by preamplification and amplification of selected target nucleic acids. Methods for analyzing RNA can entail reverse transcription of the desired RNA, followed by preamplification and amplification of selected target nucleic acids.

Abstract translation: 本发明提供从小样品甚至单细胞分析基因组DNA和/或RNA的方法。 分析基因组DNA的方法可能需要全基因组扩增（WGA），然后对选定的靶核酸进行预扩增和扩增。 分析RNA的方法可能需要所需RNA的逆转录，然后对选定的靶核酸进行预扩增和扩增。

公开(公告)号：US20150361486A1

公开(公告)日：2015-12-17

申请号：US14713887

申请日：2015-05-15

Applicant: FLUIDIGM CORPORATION

Inventor： Kenneth J. Livak ,  Stacey N. Meyers ,  Xiaohui Wang ,  Jun Wang

IPC: C12Q1/68

CPC classification number: C12Q1/6818 ,  C12Q2525/151

Abstract: The invention provides a method for detecting a target nucleotide sequence by tagging the nucleotide sequence with a nucleotide tag, providing a probe oligonucleotide with a melting temperature Tm1, comprising a regulatory sequence and a nucleotide tag recognition sequence; incorporating the probe oligonucleotide into the tagged polynucleotide in a polynucleotide amplification reaction, providing a regulatory oligonucleotide with a melting temperature Tm2, comprising a sequence segment that complementary to the regulatory sequence and a tail segment that does not hybridize to the probe nucleotide when the sequence segment and the regulatory sequence are annealed, amplifying the tagged target nucleic acid sequence in a PCR amplification reaction using the probe oligonucleotide as a primer, and using a DNA polymerase with high strand displacement activity and low 5′-nuclease activity, and detecting the amplification product; wherein Tm1 and Tm2 are higher than the annealing temperature associated with the polynucleotide amplification reaction.

Abstract translation: 本发明提供了通过用核苷酸标签标记核苷酸序列来检测靶核苷酸序列的方法，提供了具有调节序列和核苷酸标签识别序列的融解温度Tm1的探针寡核苷酸; 在多核苷酸扩增反应中将探针寡核苷酸掺入标记的多核苷酸，提供具有解链温度Tm2的调节性寡核苷酸，其包含与调节序列互补的序列区段和当序列片段不与探针核苷酸杂交时的尾段 并使用探针寡核苷酸作为引物，并使用具有高链置换活性和低5'-核酸酶活性的DNA聚合酶，在PCR扩增反应中扩增标记的靶核酸序列，并检测扩增产物 ; 其中Tm1和Tm2高于与多核苷酸扩增反应相关的退火温度。

公开(公告)号：US20150307932A1

公开(公告)日：2015-10-29

申请号：US14158618

申请日：2014-01-17

Applicant: Fluidigm Corporation

Inventor： Fatih Ozsolak ,  Patrice M. Milos

IPC: C12Q1/68

CPC classification number: C12Q1/6869 ,  C12Q1/6874 ,  C12Q2565/537 ,  C12Q2525/173 ,  C12Q2521/107

Abstract: The invention generally relates to methods for analyzing cellular nucleic acid. Methods of the invention involve capturing RNA from a lysed cell onto a substrate, producing a cDNA/RNA duplex, removing the RNA from the cDNA/RNA duplex, priming the cDNA to produce a primer/cDNA duplex, exposing the primer/cDNA duplex to at least one detectably labeled nucleotide in the presence of a polymerase capable of catalyzing addition of the nucleotide to the primer/cDNA duplex, detecting incorporation of the nucleotide into the primer portion, and repeating the exposing and detecting steps at least once.

Abstract translation: 本发明一般涉及分析细胞核酸的方法。 本发明的方法涉及将RNA从裂解细胞中捕获到底物上，产生cDNA / RNA双链体，从cDNA / RNA双链体中除去RNA，引发cDNA以产生引物/ cDNA双链体，将引物/ cDNA双链体暴露于 在能够催化向引物/ cDNA双链体加入核苷酸的聚合酶存在下，检测至少一个可检测标记的核苷酸，检测核苷酸引入引物部分，并重复曝光和检测步骤至少一次。

公开(公告)号：US09090934B2

公开(公告)日：2015-07-28

申请号：US14331112

申请日：2014-07-14

Applicant: Fluidigm Corporation

Inventor： Michael Lucero ,  Marc Unger

IPC: C12P19/34 ,  C12Q1/68 ,  G01N33/53

CPC classification number: C12Q1/686 ,  C12Q1/6804 ,  C12Q1/6813 ,  C12Q1/6876 ,  C12Q2600/118 ,  G01N33/53 ,  G01N33/6845 ,  G01N2333/435 ,  G01N2458/10

Abstract: The invention provides an assay method for detection and/or quantification of a plurality of nucleic acid or protein targets in a sample. In the method probes are used to associate a detectable tag sequence with each of the selected targets present in the sample. Probes or primers sufficient to identify at least 25, and preferably at least 500, different targets are used. The method involves segregating aliquots of the sample from each other and detecting the tag sequences in each aliquot.

Abstract translation: 本发明提供了用于检测和/或定量样品中多个核酸或蛋白质靶标的测定方法。 在该方法中，探针用于将可检测标签序列与存在于样品中的每个选定靶标相关联。 使用足以鉴定至少25个，优选至少500个不同靶标的探针或引物。 该方法包括将样品的等分试样彼此分离并检测每个等分试样中的标签序列。

公开(公告)号：US20150174575A1

公开(公告)日：2015-06-25

申请号：US14477602

申请日：2015-03-02

Applicant: Fluidigm Corporation

Inventor： Marc A. Unger ,  Lincoln McBride ,  Geoffrey Facer

IPC: B01L3/00 ,  B01L7/00 ,  C12Q1/68

CPC classification number: C12Q1/686 ,  B01L3/502738 ,  B01L7/52 ,  B01L2200/147 ,  B01L2300/0636 ,  B01L2300/0816 ,  B01L2300/0864 ,  B01L2300/0867 ,  B01L2300/0887 ,  B01L2300/0893 ,  B01L2300/123 ,  B01L2300/1805 ,  B01L2300/1827 ,  B01L2400/0481 ,  B01L2400/0655 ,  G01N27/453 ,  C12Q2565/629

Abstract: Devices and methods for performing the relative concentration of a target in a sample, the sample containing both target and non-target components, the method performed by partitioning the sample into a large number of reaction volumes such that the target is concentrated relative to the non-target, and performing a detection assay upon each reaction volume to detect the target.

Abstract translation: 用于执行样品中靶标相对浓度的装置和方法，包含靶标和非目标成分的样品，通过将样品分成大量反应体积进行的方法，使得靶相对于非目标成分集中 并且在每个反应体积上进行检测测定以检测目标。

公开(公告)号：US20150168385A1

公开(公告)日：2015-06-18

申请号：US14535162

申请日：2014-11-06

Applicant: Fluidigm Corporation

Inventor： Ian D. Manger ,  Joseph W. Barco ,  Hany R. Nassef

IPC: G01N33/53

CPC classification number: B81B3/00 ,  B01L3/5025 ,  B01L3/502707 ,  B01L3/50273 ,  B01L3/502738 ,  B01L2300/0636 ,  B01L2300/0645 ,  B01L2300/0816 ,  B01L2300/0861 ,  B01L2300/088 ,  B01L2300/0887 ,  B01L2400/0415 ,  B01L2400/0481 ,  B01L2400/0655 ,  F04B19/006 ,  F04B43/02 ,  F16K99/0001 ,  F16K99/0003 ,  F16K99/0026 ,  F16K99/0036 ,  F16K99/0046 ,  F16K99/0051 ,  F16K99/0059 ,  F16K2099/0074 ,  F16K2099/0078 ,  F16K2099/008 ,  F16K2099/0084 ,  G01N33/5304 ,  G01N33/54366 ,  Y10T436/2575

Abstract: A microfluidic device includes a plurality of first flow channels and a plurality of second flow channels, each such second flow channel intersecting multiple of the first flow channels to define intersecting volumes and a plurality of looped flow channels that each include segments of the flow channels between the intersecting volumes to define a closed loop. The microfluidic device also includes a plurality of control valves each such control valve having a control channel and a deformable segment disposed to restrict flow through a respective one of the first and second flow channels in response to an actuation force applied to the control channel to deflect the deformable segment. The microfluidic device further includes a pump operatively disposed to regulate flow through one of the looped flow channels to regulate flow by the recirculating pump.

Abstract translation: 微流体装置包括多个第一流动通道和多个第二流动通道，每个这样的第二流动通道与第一流动通道的多个相交，以限定相交的体积，以及多个环形流动通道，每个流动通道包括在 相交的卷定义一个闭环。 微流体装置还包括多个控制阀，每个这样的控制阀具有控制通道和可变形部分，可变形部分设置成响应于施加到控制通道的致动力而限制流过第一和第二流动通道中的相应一个流动通道以偏转 可变形段。 微流体装置还包括一个可操作地设置成用于调节通过环形流动通道之一的流量以调节由再循环泵流动的泵。

公开(公告)号：US20150132743A1

公开(公告)日：2015-05-14

申请号：US14477721

申请日：2014-09-04

Applicant: FLUIDIGM CORPORATION

Inventor： Camila Egidio ,  Ramesh Ramakrishnan ,  David Ruff

IPC: C12Q1/68

Abstract: Methods and reagents for detection and analysis of nucleic acids and proteins using proximity extension assays.

Abstract translation: 使用接近延伸测定法检测和分析核酸和蛋白质的方法和试剂。

公开(公告)号：US20150119298A1

公开(公告)日：2015-04-30

申请号：US14500399

申请日：2014-09-29

Applicant: Fluidigm Corporation

Inventor： Gang Sun ,  Greg Harris ,  Andy May ,  Kyle Self ,  Kevin Farrell ,  Paul Wyatt

IPC: G01N35/00

CPC classification number: G01N35/00029 ,  B01L3/5027 ,  B01L3/502715 ,  B01L2200/10 ,  B01L2300/06 ,  B01L2300/0654 ,  B01L2300/0829 ,  C12Q1/686 ,  G01N15/1475 ,  G01N21/6428 ,  G01N35/00732 ,  G01N35/00871 ,  G01N35/0092 ,  G01N35/0099 ,  G01N35/1065 ,  G01N2035/00148 ,  G01N2035/00237 ,  G01N2035/00752 ,  G01N2035/0091 ,  G01N2035/1034 ,  G01N2201/025 ,  G01N2201/0446 ,  Y10S901/01 ,  Y10T436/11

Abstract: A system for performing one or more microfluidic processes includes an integrated fluidic device comprising a plurality of well regions and a plurality of control valves and a workflow manager. The system also includes a transfer robot adapted to transfer the integrated fluidic device between a plurality of stations in response to a series of instructions from the workflow manager and a first station comprising a dispensing robot adapted to dispense at least one of a plurality of sample solutions and at least one of a plurality of reagents into the integrated fluidic device. The system further includes a second station comprising a fluidic controller unit and a third station comprising an inspection station.

Abstract translation: 用于执行一个或多个微流程过程的系统包括包括多个井区域和多个控制阀以及工作流程管理器的集成流体设备。 该系统还包括传送机器人，其适于响应于来自工作流程管理器的一系列指令而在多个站之间传送集成流体设备，第一站包括分配机器人，其适于分配多个样品溶液中的至少一个样品溶液 和多个试剂中的至少一种进入集成流体装置。 该系统还包括第二站，其包括流体控制器单元和包括检查站的第三站。

公开(公告)号：US20140318633A1

公开(公告)日：2014-10-30

申请号：US14091342

申请日：2013-11-27

Applicant: Fluidigm Corporation

Inventor： Geoffrey Facer ,  Brian Fowler ,  Emerson Cheung Quan ,  Marc Unger

IPC: F16K99/00

CPC classification number: F16K99/0015 ,  B01F5/02 ,  B01F13/0059 ,  B01L3/502707 ,  B01L3/502738 ,  B01L2200/10 ,  B01L2200/12 ,  B01L2200/16 ,  B01L2300/0627 ,  B01L2300/0816 ,  B01L2300/0874 ,  B01L2300/0887 ,  B01L2300/123 ,  B01L2400/0481 ,  B01L2400/0638 ,  B33Y80/00 ,  C12Q1/686 ,  F16K99/0026 ,  F16K99/0059 ,  F16K2099/008 ,  F16K2099/0084 ,  G01N2021/0346 ,  Y10T137/0329 ,  Y10T137/2164 ,  Y10T137/2169 ,  Y10T137/2559 ,  Y10T137/2655 ,  Y10T137/2688 ,  Y10T137/87249

Abstract: Multilevel microfluidic devices include a control line that can simultaneously actuate valves for both sample and reagent lines. Microfluidic devices are configured to contain a first reagent in a first chamber and a second reagent in a second chamber, where either or both of the first and second reagents are contained at a desired or selected pressure. Operation of a microfluidic device includes transmitting second reagent from the second chamber to the first chamber, for mixing or contact with the first reagent. Microfluidic device features such as channels, valves, chambers, can be at least partially contained, embedded, or formed by or within one or more layers or levels of an elastomeric block.

Abstract translation: 多级微流体装置包括一个控制管线，可同时为样品和试剂管线起动阀门。 微流体装置被配置为在第二室中的第一室和第二试剂中含有第一试剂，其中第一试剂和第二试剂中的任一种或两者以所需或选择的压力包含。 微流体装置的操作包括将第二试剂从第二室传输到第一室，用于与第一试剂混合或接触。 诸如通道，阀，腔室的微流体装置特征可以至少部分地包含，嵌入或由弹性体块的一个或多个层或层形成或形成。