公开(公告)号：WO2012162267A2

公开(公告)日：2012-11-29

申请号：PCT/US2012/038894

申请日：2012-05-21

Applicant: FLUIDIGM CORPORATION ,  ANDERSON, Megan ,  CHEN, Peilin ,  FOWLER, Brian ,  JONES, Robert C. ,  KAPER, Fiona ,  LEBOFSKY, Ronald ,  MAY, Andrew

Inventor： ANDERSON, Megan ,  CHEN, Peilin ,  FOWLER, Brian ,  JONES, Robert C. ,  KAPER, Fiona ,  LEBOFSKY, Ronald ,  MAY, Andrew

IPC: C40B20/00 ,  C40B50/06 ,  C40B40/06 ,  C40B30/04 ,  C12P19/34 ,  C12Q1/68

CPC classification number: C12Q1/6855 ,  C12N15/10 ,  C12N15/1065 ,  C12N15/65 ,  C12N15/66 ,  C12Q1/686 ,  C12Q1/6874

Abstract: Described herein are methods useful for incorporating one or more adaptors and/or nucleotide tag(s) and/or barcode nucleotide sequence(s) one, or typically more, target nucleotide sequences. In particular embodiments, nucleic acid fragments having adaptors, e.g., suitable for use in high-throughput DNA sequencing are generated. In other embodiments, information about a reaction mixture is encoded into a reaction product. Also described herein are methods and kits useful for amplifying one or more target nucleic acids in preparation for applications such as bidirectional nucleic acid sequencing. In particular embodiments, methods of the invention entail additionally carrying out bidirectional DNA sequencing. Also described herein are methods for encoding and detecting and/or quantifying alleles by primer extension.

Abstract translation: 本文所描述的方法可用于掺入一个或多个衔接子和/或核苷酸标签和/或条形码核苷酸序列，一个或多个靶核苷酸序列。 在具体实施方案中，产生具有适配器（例如适合用于高通量DNA测序）的核酸片段。 在其它实施方案中，关于反应混合物的信息被编码成反应产物。 本文还描述了可用于扩增一种或多种靶核酸以准备应用如双向核酸测序的方法和试剂盒。 在具体实施方案中，本发明的方法还需要进行双向DNA测序。 本文还描述了通过引物延伸来编码和检测和/或定量等位基因的方法。

公开(公告)号：WO2021168439A1

公开(公告)日：2021-08-26

申请号：PCT/US2021/019101

申请日：2021-02-22

Applicant: FLUIDIGM CORPORATION

Inventor： PHELAN, Michael L. ,  KUBU, Christopher J. ,  FOWLER, Brian ,  SUN, Gang ,  PATEL, Nikita ,  RAMALINGAM, Naveen

IPC: C12P19/34 ,  B01F11/00 ,  B01F11/02

Abstract: Described herein are methods, kits and systems for sample enrichment, multi-step library preparation, sample normalization, detection of sample biomolecules and combinations thereof. Enrichment and multi-step library preparation is described in the context of microfluidic workflows. Sample barcoding methods and kits are described for increasing sample throughput while reducing background in negative samples. Integrated microfluidic devices comprising sample processing unit cells coupled to an array of reaction sites are provided for integrated workflows.

公开(公告)号：WO2010077618A1

公开(公告)日：2010-07-08

申请号：PCT/US2009/067037

申请日：2009-12-07

Applicant: FLUIDIGM CORPORATION ,  FOWLER, Brian

Inventor： FOWLER, Brian

IPC: B01L99/00

CPC classification number: B01L3/502784 ,  B01J2219/00286 ,  B01J2219/00317 ,  B01J2219/00398 ,  B01J2219/00418 ,  B01L7/525 ,  B01L2200/027 ,  B01L2300/0816 ,  B01L2300/089 ,  B01L2300/1805 ,  B01L2400/0427

Abstract: A microfluidic device includes a pressure source and a control line in fluid communication with the pressure source. The microfluidic device also includes a plurality of valves operated via the control line and an independent valve positioned adjacent the control line and between the pressure source and the plurality of valves.

Abstract translation: 微流体装置包括压力源和与压力源流体连通的控制管线。 微流体装置还包括经由控制管线操作的多个阀和邻近控制管线定位并且在压力源和多个阀之间的独立阀。

公开(公告)号：WO2013130714A1

公开(公告)日：2013-09-06

申请号：PCT/US2013/028170

申请日：2013-02-28

Applicant: FLUIDIGM CORPORATION ,  FOWLER, Brian ,  KIMBALL, Jake ,  MAUNG, Myo Thu ,  MAY, Andrew ,  NORRIS, Michael C. ,  TOPPANI, Dominique G. ,  UNGER, Marc A. ,  WANG, Jing ,  WEST, Jason A. A.

Inventor： FOWLER, Brian ,  KIMBALL, Jake ,  MAUNG, Myo Thu ,  MAY, Andrew ,  NORRIS, Michael C. ,  TOPPANI, Dominique G. ,  UNGER, Marc A. ,  WANG, Jing ,  WEST, Jason A. A.

IPC: G01N15/10

CPC classification number: G01N1/28 ,  B01L3/502761 ,  B01L7/52 ,  B01L2200/0668 ,  B01L2300/0864 ,  B01L2400/0409 ,  B01L2400/0415 ,  B01L2400/043 ,  B01L2400/0487 ,  B01L2400/086 ,  B01L2400/088 ,  C12P19/34 ,  C12Q1/6813 ,  C12Q1/6844 ,  C12Q1/686 ,  C12Q1/6869 ,  G01N1/34 ,  G01N15/1484 ,  C12Q2565/629

Abstract: Methods, systems, and devices are described for multiple single-cell capturing and processing utilizing microfluidics. Tools and techniques are provided for capturing, partitioning, and/or manipulating individual cells from a larger population of cells along with generating genetic information and/or reactions related to each individual cell. Different capture configurations may be utilized to capture individual cells and then processing each individual cell in a multi-chamber reaction configuration. Some embodiments may provide for specific target amplification, whole genome amplification, whole transcriptome amplification, real-time PCR preparation, copy number variation, preamplification, mRNA sequencing, and/or haplotyping of the multiple individual cells that have been partitioned from the larger population of cells. Some embodiments may provide for other applications. Some embodiments may be configured for imaging the individual cells or associated reaction products as part of the processing. Reaction products may be harvested and/or further analyzed in some cases.

Abstract translation: 描述了利用微流体的多单细胞捕获和处理的方法，系统和装置。 提供的工具和技术用于捕获，分配和/或操纵来自较大群体的细胞的各个细胞以及产生与每个单个细胞相关的遗传信息和/或反应。 可以使用不同的捕获配置来捕获单个细胞，然后在多室反应配置中处理每个单独的细胞。 一些实施方案可以提供已经从较大群体中划分的多个单个细胞的特异性靶扩增，全基因组扩增，全转录组扩增，实时PCR制备，拷贝数变异，预扩增，mRNA测序和/或单倍型 细胞。 一些实施例可以提供其他应用。 一些实施例可以被配置为用于对作为处理的一部分的各个单元或相关联的反应产物进行成像。 在某些情况下可以收获和/或进一步分析反应产物。

公开(公告)号：WO2009126826A1

公开(公告)日：2009-10-15

申请号：PCT/US2009/040104

申请日：2009-04-09

Applicant: FLUIDIGM CORPORATION ,  FACER, Geoffrey ,  FOWLER, Brian ,  QUAN, Emerson, Cheung ,  UNGER, Marc

Inventor： FACER, Geoffrey ,  FOWLER, Brian ,  QUAN, Emerson, Cheung ,  UNGER, Marc

IPC: B81B3/00

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。

公开(公告)号：WO2003085379A3

公开(公告)日：2003-10-16

申请号：PCT/US2003/009997

申请日：2003-04-01

Applicant: FLUIDIGM CORPORATION ,  CHOU, Hou-Pu ,  DARIDON, Antoine ,  FARRELL, Kevin ,  FOWLER, Brian ,  HAO, Cunsheng (Casey) ,  JAVADI, Shervin ,  LIAU, Yish-Hann ,  MANGER, Ian, D. ,  NASSEF, Hany, Ramez ,  NORTON, Pierce ,  THRONDSET, William

Inventor： CHOU, Hou-Pu ,  DARIDON, Antoine ,  FARRELL, Kevin ,  FOWLER, Brian ,  HAO, Cunsheng (Casey) ,  JAVADI, Shervin ,  LIAU, Yish-Hann ,  MANGER, Ian, D. ,  NASSEF, Hany, Ramez ,  NORTON, Pierce ,  THRONDSET, William

IPC: C12M1/34

Abstract: The invention provides systems (2000), including microfluidic mechanisms, methods, and kits, for the microfluidic manipulation and/or detection of particles, such as cells and/or beads. These mechanisms may enable controlled input, movement/positioning, retention/localization, treatment, measurement, release, and/or output of particles. Furthermore, these mechanisms may be combined in any suitable order and/or employed for any number of suitable times in the system to allow particles to be sorted, cultured, mixed, treated, and/or assayed, among others. These combinations may allow the response of particles to treatment to be measured on a shorter time scale than was previously possible. Therefore, systems of the invention may allow a broad range of cell and particle assays, such as drug screens, cell characterizations, research studies, and/or clinical analyses, among others, to be scaled down to microfluidic size. Such scaled-down assays may use less sample and reagent, may be less labor intensive, and/or may be more informative than comparable macrofluidic assays.

公开(公告)号：WO2015172080A1

公开(公告)日：2015-11-12

申请号：PCT/US2015/029986

申请日：2015-05-08

Applicant: FLUIDIGM CORPORATION

Inventor： WEST, Jason A. A. ,  FOWLER, Brian ,  CHARN, Tze-Howe ,  JOHNSON, Christian F. ,  UNGER, Marc A.

IPC: C12Q1/68

CPC classification number: C12Q1/6855 ,  C12N15/1065 ,  C12Q1/6806 ,  C12Q2565/501 ,  C12Q2521/301 ,  C12Q2521/501 ,  C12Q2525/155 ,  C12Q2537/162 ,  C12Q2565/543 ,  C12Q2535/122

Abstract: This disclosure provides a method of forming tagged nucleic acid sequences. A target polynucleotide is immobilized on a solid support; a recognition-oligonucleotide is hybridized thereto; the recognition-oligonucleotide-target polynucleotide hybrid is cleaved; and an adapter nucleic acid is ligated to the cleaved target polynucleotide, thereby forming a tagged nucleic acid sequence. Also provided is a method of forming a tagged single stranded cDNA; a method of forming a plurality of tagged heterogeneous nucleic acid sequences; a library of recognition-oligonucleotides; and methods for amplifying a cDNA sequence immobilized on a solid support. These methods and products can be used alone or in combination for integrated single cell sequencing, and can be adapted for use in a microfluidic apparatus or device.

Abstract translation: 本公开提供了形成标记的核酸序列的方法。 靶多核苷酸固定在固体支持物上; 识别寡核苷酸与其杂交; 识别寡核苷酸靶多核苷酸杂交体被切割; 并将衔接子核酸连接到切割的靶多核苷酸上，从而形成标记的核酸序列。 还提供了形成标记的单链cDNA的方法; 形成多个标记的异源核酸序列的方法; 识别寡核苷酸文库; 以及用于扩增固定在固体支持物上的cDNA序列的方法。 这些方法和产品可以单独使用或组合使用以用于集成的单细胞测序，并且可以适用于微流体装置或装置。

公开(公告)号：WO2014144789A2

公开(公告)日：2014-09-18

申请号：PCT/US2014/029344

申请日：2014-03-14

Applicant: FLUIDIGM CORPORATION

Inventor： WEST, Jason A. A. ,  FOWLER, Brian

IPC: C12Q1/00

CPC classification number: G01N35/08 ,  B01L3/502738 ,  B01L2200/027 ,  B01L2200/0652 ,  B01L2200/0668 ,  B01L2200/10 ,  B01L2300/0816 ,  B01L2300/0864 ,  B01L2300/0867 ,  B01L2300/10 ,  B01L2300/12 ,  B01L2400/0487 ,  C12M23/24 ,  C12M29/04 ,  G01N15/1056 ,  G01N15/1484 ,  G01N33/5005 ,  G01N33/5097 ,  G01N33/56961 ,  G01N33/56966 ,  G01N2015/1081

Abstract: Methods for cell analysis are provided, comprising cell capturing, characterization, transport, and culture. In an exemplary method individual cells (and/or cellular units) are flowed into a microfluidic channel, the channel is partitioned into a plurality of contiguous segments, capturing at least one cell in at least one segment, A characteristic of one or more captured cells is determined and the cell(s) and combinations of cells are transported to specified cell holding chamber(s) based on the determined characteristic(s). Also provided are devices and systems for cell analysis.

Abstract translation: 提供了用于细胞分析的方法，包括细胞捕获，表征，转运和培养。 在一个示例性方法中，个体细胞（和/或细胞单位）流入微流体通道，通道被划分成多个连续区段，捕获至少一个区段中的至少一个细胞，一个或多个捕获细胞的特征 并且基于所确定的特性将单元和单元的组合输送到指定的单元容纳室。 还提供了用于细胞分析的装置和系统。

公开(公告)号：WO2013177206A2

公开(公告)日：2013-11-28

申请号：PCT/US2013/042086

申请日：2013-05-21

Applicant: FLUIDIGM CORPORATION ,  ANDERSON, Megan ,  CHEN, Peilin ,  FOWLER, Brian ,  KAPER, Fiona ,  LEBOFSKY, Ronald ,  MAY, Andrew

Inventor： ANDERSON, Megan ,  CHEN, Peilin ,  FOWLER, Brian ,  KAPER, Fiona ,  LEBOFSKY, Ronald ,  MAY, Andrew

IPC: C12Q1/68

CPC classification number: C12Q1/6813 ,  B01D15/08 ,  C12Q1/6806 ,  C12Q2563/159 ,  C12Q2563/179 ,  C12Q2565/629

Abstract: In certain embodiments, the invention provides methods and devices for assaying single particles in a population of particles, wherein at least two parameters are measured for each particle. One or more parameters can be measured while the particles are in the separate reaction volumes. Alternatively or in addition, one or more parameters can be measured in a later analytic step, e.g., where reactions are carried out in the separate reaction volumes and the reaction products are recovered and analyzed. In particular embodiments, one or more parameter measurements are carried out "in parallel," i.e., essentially simultaneously in the separate reaction volumes.

Abstract translation: 在某些实施方案中，本发明提供用于测定颗粒群体中的单个颗粒的方法和装置，其中针对每个颗粒测量至少两个参数。 当颗粒处于分开的反应体积中时，可以测量一个或多个参数。 或者或另外，可以在稍后的分析步骤中测量一个或多个参数，例如其中在分开的反应体积中进行反应并回收和分析反应产物。 在具体实施方案中，“并行地”执行一个或多个参数测量，即基本上同时在单独的反应体积中进行。

公开(公告)号：WO2011040884A2

公开(公告)日：2011-04-07

申请号：PCT/SG2010/000369

申请日：2010-09-29

Applicant: FLUIDIGM CORPORATION ,  COHEN, David ,  MAY, Andrew ,  PIEPRZYK, Martin ,  FOWLER, Brian ,  LEE, Kim Huat ,  YAN, Jun ,  ZHOU, Ming Fang ,  NG, Seng Beng

Inventor： COHEN, David ,  MAY, Andrew ,  PIEPRZYK, Martin ,  FOWLER, Brian ,  LEE, Kim Huat ,  YAN, Jun ,  ZHOU, Ming Fang ,  NG, Seng Beng

IPC: G01N35/08 ,  G01N33/48

CPC classification number: B01L3/5027 ,  B01L3/502738 ,  B01L7/52 ,  B01L9/527 ,  B01L2200/12 ,  B01L2300/045 ,  B01L2300/0887 ,  B01L2300/123 ,  B01L2400/0655 ,  G01N1/405

Abstract: The present invention includes microfluidic systems having a microfabricated cavity that may be covered with a removable cover, where the removable cover allows at least part of the opening of the microfabricated cavity to be exposed or directly accessed by an operator. The microfluidic systems comprise chambers, flow and control channels formed in elastomeric layers that may comprise PDMS. The removable cover comprises a thermoplastic base film bonded to an elastomer layer by an adhesive layer. When the removable cover is peeled off, the chamber is at least partially open to allow sample extraction from the chamber. The chamber may have macromolecular crystals formed inside or resulting contents from a PCR reaction. The invention also includes a method for making vias in elastomeric layers by using the removable cover. The invention further includes methods and devices for peeling the peelable cover or a removable component such as Integrated Heater Spreader.

Abstract translation: 本发明包括具有微型空腔的​​微流体系统，其可以被可移除的盖覆盖，其中可移除的盖允许微加工腔的至少一部分开口被操作者暴露或直接访问。 微流体系统包括在可包括PDMS的弹性体层中形成的腔室，流动和控制通道。 可移除的盖包括通过粘合剂层结合到弹性体层的热塑性基底膜。 当可移除的盖被剥离时，室至少部分地打开以允许从室中取样。 该室可能在PCR反应内部形成大分子晶体或产生内含物。 本发明还包括通过使用可移除盖制造弹性体层中的通孔的方法。 本发明还包括用于剥离可剥离盖或诸如集成式加热器撒布机的可移除部件的方法和装置。