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：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：EP3285062A1

公开(公告)日：2018-02-21

申请号：EP17180284.6

申请日：2013-02-28

Applicant: Fluidigm Corporation

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: 描述了使用微流体的多个单细胞捕获和处理的方法，系统和设备。 提供了用于捕获，分配和/或操纵来自更大细胞群体的个体细胞以及产生与每个个体细胞相关的遗传信息和/或反应的工具和技术。 可以使用不同的捕获配置来捕获单个细胞，然后在多腔室反应配置中处理每个单独的细胞。 一些实施例可以提供特定目标扩增，全基因组扩增，全转录扩增，实时PCR制备，拷贝数变异，预扩增的mRNA的测序，和/或单倍型分析已经从较大人口划分的多个单个细胞的 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用于将单独的单元或相关联的反应产物成像为处理的一部分。 反应产物可能在一些情况下收获和/或进一步分析。

公开(公告)号：EP2970849B1

公开(公告)日：2019-08-21

申请号：EP14763560.1

申请日：2014-03-14

Applicant: Fluidigm Corporation

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

IPC: C12M1/22 ,  G01N33/50 ,  G01N33/569

公开(公告)号：EP3627140A1

公开(公告)日：2020-03-25

申请号：EP19189921.0

申请日：2013-02-28

Applicant: Fluidigm Corporation

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 ,  C12Q1/6806

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.

公开(公告)号：EP2820394B1

公开(公告)日：2017-07-12

申请号：EP13754093.6

申请日：2013-02-28

Applicant: Fluidigm Corporation

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.

公开(公告)号：EP2970849A2

公开(公告)日：2016-01-20

申请号：EP14763560.1

申请日：2014-03-14

Applicant: Fluidigm Corporation

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

IPC: C12M1/22

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，运输和培养。 在示例性方法中，将单个细胞（和/或细胞单位）流入微流体通道，将该通道划分成多个连续片段，在至少一个片段中捕获至少一个细胞。一个或多个捕获细胞的特征 并且基于确定的特征将细胞和细胞的组合传输到指定的细胞容纳室。 还提供了用于细胞分析的装置和系统。

公开(公告)号：EP3285062B1

公开(公告)日：2019-08-07

申请号：EP17180284.6

申请日：2013-02-28

Applicant: Fluidigm Corporation

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

公开(公告)号：EP3140428A1

公开(公告)日：2017-03-15

申请号：EP15789313.2

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，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。