公开(公告)号：US09952126B2

公开(公告)日：2018-04-24

申请号：US13781313

申请日：2013-02-28

Applicant: Fluidigm Corporation

Inventor： Brian Fowler ,  Jake Kimball ,  Myo Thu Maung ,  Andrew May ,  Michael C Norris ,  Dominique Toppani ,  Marc A. Unger ,  Jing Wang ,  Jason A. A. West

IPC: C12P19/34 ,  G01N1/28 ,  C12Q1/68 ,  G01N1/34 ,  G01N15/14 ,  B01L3/00 ,  B01L7/00

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：US09643178B2

公开(公告)日：2017-05-09

申请号：US13439364

申请日：2012-04-04

Applicant: Marc Unger ,  Ian D. Manger ,  Michael Lucero ,  Yong Yi ,  Emily Miyashita-Lin ,  Anja Wienecke ,  Geoffrey Facer

Inventor： Marc Unger ,  Ian D. Manger ,  Michael Lucero ,  Yong Yi ,  Emily Miyashita-Lin ,  Anja Wienecke ,  Geoffrey Facer

IPC: B01L3/00 ,  C12Q1/68 ,  G01N35/00 ,  B01F13/00 ,  C40B60/14

CPC classification number: C12M1/38 ,  B01F13/0059 ,  B01J2219/00337 ,  B01J2219/00353 ,  B01J2219/00403 ,  B01J2219/00495 ,  B01J2219/00585 ,  B01J2219/0059 ,  B01J2219/0072 ,  B01L3/502707 ,  B01L3/502715 ,  B01L3/50851 ,  B01L2300/0627 ,  B01L2300/123 ,  B01L2400/06 ,  C12Q1/68 ,  C12Q1/6844 ,  C40B60/14 ,  F16K11/20 ,  G01N35/00029 ,  G01N35/0098 ,  G01N2035/00366 ,  Y10T137/2076 ,  Y10T137/2496 ,  Y10T137/86292 ,  Y10T137/87249 ,  Y10T436/2575 ,  C12Q2565/501

Abstract: A variety of elastomeric-based microfluidic devices and methods for using and manufacturing such devices are provided. Certain of the devices have arrays of reaction sites to facilitate high throughput analyzes. Some devices also include reaction sites located at the end of blind channels at which reagents have been previously deposited during manufacture. The reagents become suspended once sample is introduced into the reaction site. The devices can be utilized with a variety of heating devices and thus can be used in a variety of analyzes requiring temperature control, including thermocycling applications such as nucleic acid amplification reactions, genotyping and gene expression analyzes.

公开(公告)号：US09623413B2

公开(公告)日：2017-04-18

申请号：US12761917

申请日：2010-04-16

Applicant: Geoffrey Facer ,  Robert Grossman ,  Marc Unger ,  Phillip Lam ,  Hou-Pu Chou ,  Jake Kimball ,  Martin Pieprzyk ,  Antoine Daridon

Inventor： Geoffrey Facer ,  Robert Grossman ,  Marc Unger ,  Phillip Lam ,  Hou-Pu Chou ,  Jake Kimball ,  Martin Pieprzyk ,  Antoine Daridon

IPC: B01L3/00 ,  B01L7/00 ,  B01L9/00 ,  C30B7/00 ,  C30B29/58 ,  C30B35/00 ,  B01L3/02 ,  B01L3/06

CPC classification number: C12M1/00 ,  B01J2219/00317 ,  B01J2219/00495 ,  B01L3/0293 ,  B01L3/06 ,  B01L3/5025 ,  B01L3/502707 ,  B01L3/502715 ,  B01L3/50273 ,  B01L3/502738 ,  B01L3/5635 ,  B01L3/565 ,  B01L7/52 ,  B01L9/527 ,  B01L2200/027 ,  B01L2200/0605 ,  B01L2200/10 ,  B01L2300/0816 ,  B01L2300/0829 ,  B01L2300/0874 ,  B01L2300/0887 ,  B01L2300/14 ,  B01L2300/1805 ,  B01L2400/0481 ,  B01L2400/0487 ,  B01L2400/0605 ,  B01L2400/0655 ,  C30B7/00 ,  C30B29/58 ,  Y10T29/49826 ,  Y10T436/25

Abstract: Methods and systems are provided for conducting a reaction at a selected temperature or range of temperatures over time. An array device is provided. The array device contains separate reaction chambers and is formed as an elastomeric block from multiple layers. At least one layer has at least one recess that recess has at least one deflectable membrane integral to the layer with the recess. The array device has a thermal transfer device proximal to at least one of the reaction chambers. The thermal transfer device is formed to contact a thermal control source. Reagents for carrying out a desired reaction are introduced into the array device. The array device is contacted with a thermal control device such that the thermal control device is in thermal communication with the thermal control source so that a temperature of the reaction in at least one of the reaction chamber is changed as a result of a change in temperature of the thermal control source.

公开(公告)号：US09593698B2

公开(公告)日：2017-03-14

申请号：US14072243

申请日：2013-11-05

Applicant: Fluidigm Corporation

Inventor： David Fernandes ,  Hou-Pu Chou ,  Marc A. Unger

IPC: F16C3/00 ,  F16K99/00 ,  B01L3/00 ,  F15C3/00 ,  F15C3/04 ,  G06F17/50 ,  H01H29/00

CPC classification number: F15C3/00 ,  A61M2206/22 ,  B01L3/502707 ,  B01L3/50273 ,  B01L3/502738 ,  B01L2200/0621 ,  B01L2200/14 ,  B01L2300/123 ,  B01L2400/0487 ,  B01L2400/0638 ,  B01L2400/0655 ,  F15C3/04 ,  F16K99/0001 ,  F16K99/0009 ,  F16K99/0015 ,  F16K99/0034 ,  F16K99/0059 ,  F16K2099/0074 ,  F16K2099/008 ,  F16K2099/0082 ,  F16K2099/0084 ,  G06F17/509 ,  G06F2217/16 ,  H01H2029/008 ,  Y10T137/206 ,  Y10T137/2202 ,  Y10T137/2218 ,  Y10T137/2224 ,  Y10T137/7879 ,  Y10T137/7891 ,  Y10T137/7892

Abstract: A microfabricated fluidic unidirectional valve includes a microfabricated elastomer material having a flow through channel. The microfabricated fluidic unidirectional valve also includes an elastomer flap attached to the elastomer material in the flow through channel. The elastomer flap forms a seal in the flow through channel to prevent fluid from flowing in a first direction through the flow through channel and to allow fluid flow in a second direction through the flow through channel.

Abstract translation: 微制造的流体单向阀包括具有流通通道的微加工弹性体材料。 微制造的流体单向阀还包括在流动通道中附接到弹性体材料的弹性体翼片。 弹性翼片在流通通道中形成密封，以防止流体沿第一方向流过通过通道并允许流体沿第二方向流过通过通道的流体。

公开(公告)号：US09498776B2

公开(公告)日：2016-11-22

申请号：US14720305

申请日：2015-05-22

Applicant: Fluidigm Corporation

Inventor： David Cohen ,  Andrew May ,  Martin Pieprzyk ,  Brian Fowler ,  Kim Huat Lee ,  Jun Yan ,  Ming Fang Zhou ,  Seng Beng Ng

IPC: B01L99/00 ,  B01L3/00 ,  B01L9/00 ,  B01L7/00 ,  G01N1/40

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.

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：US20150273735A1

公开(公告)日：2015-10-01

申请号：US13813625

申请日：2009-07-23

Applicant: David S. Cohen

Inventor： David S. Cohen

IPC: B29C41/02 ,  B32B38/00 ,  B01L3/00 ,  B32B37/18

CPC classification number: B29C41/02 ,  B01L3/502 ,  B01L3/502707 ,  B01L2300/0861 ,  B01L2300/123 ,  B01L2400/0481 ,  B01L2400/0655 ,  B29K2083/00 ,  B29L2009/00 ,  B29L2031/755 ,  B32B37/182 ,  B32B38/0008 ,  B32B2309/105 ,  B32B2383/00 ,  G01N30/6095

Abstract: An integrated fluidic chip includes a substrate defined by a lateral surface area greater than 28 square inches. The integrated fluidic chip also includes a first elastomeric layer having a mold surface and a top surface. The mold surface of the first elastomeric layer is joined to a portion of the substrate. The first elastomeric layer includes a plurality of first channels extending normally from the substrate to a first dimension inside the first elastomeric layer. The integrated fluidic chip further includes a second elastomeric layer having a mold surface and a top surface. The mold surface of the second elastomeric layer is joined to at least a portion of the top surface of the first elastomeric layer.

Abstract translation: 集成流体芯片包括由大于28平方英寸的侧表面积限定的基板。 集成流体芯片还包括具有模具表面和顶表面的第一弹性体层。 第一弹性体层的模具表面连接到基底的一部分。 第一弹性体层包括多个第一通道，其从基底正常延伸到第一弹性体层内的第一尺寸。 集成流体芯片还包括具有模具表面和顶表面的第二弹性体层。 第二弹性体层的模具表面连接到第一弹性体层的顶表面的至少一部分。

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。