公开(公告)号：US09926521B2

公开(公告)日：2018-03-27

申请号：US14188660

申请日：2014-02-24

Applicant: Fluidigm Corporation

Inventor： Antoine Daridon

IPC: C12M3/00 ,  B01L3/00 ,  C12M1/34 ,  C12M3/06 ,  G01N33/483 ,  G02B21/32 ,  G01N15/14 ,  G01N15/00 ,  G01N15/02

CPC classification number: C12M1/34 ,  B01L3/502738 ,  B01L3/502746 ,  B01L3/502753 ,  B01L3/502761 ,  B01L2200/0636 ,  B01L2200/0647 ,  B01L2200/0652 ,  B01L2200/0668 ,  B01L2200/10 ,  B01L2200/12 ,  B01L2200/16 ,  B01L2300/06 ,  B01L2300/0636 ,  B01L2300/0645 ,  B01L2300/0681 ,  B01L2300/0861 ,  B01L2300/0867 ,  B01L2300/087 ,  B01L2300/088 ,  B01L2300/0887 ,  B01L2300/0893 ,  B01L2300/123 ,  B01L2400/0409 ,  B01L2400/0415 ,  B01L2400/0481 ,  B01L2400/0622 ,  B01L2400/0655 ,  C12M21/06 ,  C12M23/16 ,  G01N15/1484 ,  G01N33/4833 ,  G01N2015/008 ,  G01N2015/0288 ,  G01N2015/149 ,  G01N2015/1493 ,  G02B21/32

Abstract: The invention provides systems, including apparatus, methods, and kits, for the microfluidic manipulation and/or detection of particles, such as cells and/or beads. The invention provides systems, including apparatus, methods, and kits, for the microfluidic manipulation and/or analysis of particles, such as cells, viruses, organelles, beads, and/or vesicles. The invention also provides microfluidic mechanisms for carrying out these manipulations and analyses. 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 suitable number of times within a system. Accordingly, these combinations may allow particles to be sorted, cultured, mixed, treated, and/or assayed, among others, as single particles, mixed groups of particles, arrays of particles, heterogeneous particle sets, and/or homogeneous particle sets, among others, in series and/or in parallel. In addition, these combinations may enable microfluidic systems to be reused. Furthermore, 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：US09644231B2

公开(公告)日：2017-05-09

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: G01N21/64 ,  C40B30/04 ,  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.

公开(公告)号：US09587272B2

公开(公告)日：2017-03-07

申请号：US14340446

申请日：2014-07-24

Applicant: Fluidigm Corporation

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

IPC: C07H21/04 ,  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 is at least partially complementary to the regulatory sequence, amplifying the tagged target nucleic acid sequence in a PCR amplification reaction using the probe oligonucleotide as a primer, 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，提供了包含调节序列和核苷酸标签识别序列的具有解链温度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中，提供具有解链温度Tm2的调节性寡核苷酸，其包含与调节序列至少部分互补的序列片段，在PCR扩增中扩增标记的靶核酸序列 使用探针寡核苷酸作为引物的反应，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：US09506914B2

公开(公告)日：2016-11-29

申请号：US14535162

申请日：2014-11-06

Applicant: Fluidigm Corporation

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

IPC: B81B3/00 ,  G01N33/53 ,  F04B19/00 ,  F04B43/02 ,  F16K99/00 ,  G01N33/543 ,  B01L3/00

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：US09506845B2

公开(公告)日：2016-11-29

申请号：US13781318

申请日：2013-02-28

Applicant: Fluidigm Corporation

Inventor： Brian Fowler ,  Jake Kimball ,  Myo Thu Maung ,  Andrew May ,  Michael C. Norris ,  Dominique G. 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：US09440231B2

公开(公告)日：2016-09-13

申请号：US12673192

申请日：2008-08-08

Applicant: Jason A.A. West ,  Jesse Thompson

Inventor： Jason A.A. West ,  Jesse Thompson

IPC: B01L3/00 ,  G01N30/00 ,  B29C45/03 ,  B29C59/02 ,  B29C45/00 ,  B29C45/73 ,  B05D5/00 ,  B32B37/06 ,  G01N33/543 ,  B29L31/00

CPC classification number: B01L3/502707 ,  B01J19/0046 ,  B01J2219/00596 ,  B01J2219/00605 ,  B01J2219/00608 ,  B01J2219/00722 ,  B01L2200/0636 ,  B01L2200/0647 ,  B01L2200/0668 ,  B01L2200/0689 ,  B01L2300/0636 ,  B01L2300/0681 ,  B01L2300/0816 ,  B01L2300/0819 ,  B01L2300/0887 ,  B01L2300/12 ,  B29L2031/756 ,  G01N33/54386

Abstract: Provided are microfluidic devices and methods for fabricating and bonding such devices. Also provided are kits for analyzing analyte-containing samples and for lysing cells.

Abstract translation: 提供了用于制造和结合这些装置的微流体装置和方法。 还提供了用于分析含分析物的样品和裂解细胞的试剂盒。

公开(公告)号：US09381512B2

公开(公告)日：2016-07-05

申请号：US14071598

申请日：2013-11-04

Applicant: Fluidigm Corporation

Inventor： David S. Cohen ,  Jing Wang ,  Andrew May ,  Robert C. Jones ,  Hany Nassef

IPC: B01L3/00 ,  F16K99/00 ,  B01L7/00

CPC classification number: B01L3/502738 ,  B01L3/5025 ,  B01L7/52 ,  B01L2200/0684 ,  B01L2300/0816 ,  B01L2300/0864 ,  B01L2300/087 ,  B01L2300/0874 ,  B01L2300/0887 ,  B01L2300/123 ,  B01L2400/0487 ,  B01L2400/0605 ,  B01L2400/0638 ,  B01L2400/0655 ,  B33Y80/00 ,  F16K99/0001 ,  F16K99/0026 ,  F16K99/0059 ,  F16K2099/0074 ,  F16K2099/0078 ,  F16K2099/008 ,  F16K2099/0084 ,  Y10T137/0318 ,  Y10T137/85938

Abstract: New high density microfluidic devices and methods provide precise metering of fluid volumes and efficient mixing of the metered volumes. A first solution is introduced into a segment of a flow channel in fluidic communication with a reaction chamber. A second solution is flowed through the segment so that the first solution is displaced into the reaction chamber, and a volume of the second solution enters the chamber. The chamber can then be isolated and reactions within the chamber can be initiated and/or detected. High throughput methods of genetic analysis can be carried out with greater accuracy than previously available.

公开(公告)号：US09205468B2

公开(公告)日：2015-12-08

申请号：US13516146

申请日：2010-11-30

Applicant: Jing Wang ,  Timothy M. Woudenberg

Inventor： Jing Wang ,  Timothy M. Woudenberg

IPC: B08B9/027 ,  B01L3/00 ,  B01L99/00 ,  F16K99/00

CPC classification number: B08B9/027 ,  B01L3/5027 ,  B01L99/00 ,  B01L2300/123 ,  B01L2400/0481 ,  B01L2400/0655 ,  F16K99/0001 ,  F16K99/0026 ,  F16K99/0059 ,  F16K2099/0074 ,  F16K2099/0078 ,  F16K2099/008 ,  F16K2099/0084

Abstract: A method for rendering a microfluidic device suitable for reuse for nucleic acid analysis is provided. The method may include flowing a nucleic acid inactivating solution into a microfluidic channel of the device by pumping; and then flowing a wash solution into the channel by pumping, thereby displacing the nucleic acid inactivating solution from the channel, whereby any residual nucleic acid from a prior use of the device is inactivated.

Abstract translation: 提供了一种用于制备适于重新用于核酸分析的微流体装置的方法。 该方法可以包括通过泵送将核酸灭活溶液流动到装置的微流体通道中; 然后通过泵送将洗涤溶液流入通道，​​从而使来自通道的核酸灭活溶液移位，由此来自先前使用该装置的任何残留核酸被灭活。

公开(公告)号：US20140308178A1

公开(公告)日：2014-10-16

申请号：US14091344

申请日：2013-11-27

Applicant: Fluidigm Corporation

Inventor： Tim Woudenberg ,  Jing Wang ,  Hou-Pu Chou

IPC: G01N1/28

CPC classification number: B01L3/502715 ,  B01F13/0059 ,  B01F13/0094 ,  B01F13/1013 ,  B01F13/1022 ,  B01J2219/00317 ,  B01J2219/00479 ,  B01J2219/00585 ,  B01L3/5025 ,  B01L3/502723 ,  B01L3/502738 ,  B01L3/502746 ,  B01L7/52 ,  B01L2200/027 ,  B01L2200/0605 ,  B01L2300/0829 ,  B01L2300/0861 ,  B01L2300/0864 ,  B01L2300/0867 ,  B01L2300/0877 ,  B01L2300/0887 ,  B01L2300/123 ,  B01L2300/1805 ,  B01L2400/0487 ,  B01L2400/0605 ,  B01L2400/0655 ,  B01L2400/082 ,  B33Y80/00 ,  G01N1/28 ,  H04W4/70 ,  H04W76/30

Abstract: Microfluidic devices are described that include a rigid base layer, and an elastomeric layer on the base layer. The elastomeric layer may include at least part of a fluid channel for transporting a liquid reagent, and a vent channel that accepts gas diffusing through the elastomeric layer from the flow channel and vents it out of the elastomeric layer. The devices may also include a mixing chamber fluidly connected to the fluid channel, and a control channel overlapping with a deflectable membrane that defines a portion of the flow channel, where the control channel may be operable to change a rate at which the liquid reagent flows through the fluid channel. The devices may further include a rigid plastic layer on the elastomeric layer.

Abstract translation: 描述了包括刚性基层和基层上的弹性体层的微流体装置。 弹性体层可以包括用于输送液体试剂的流体通道的至少一部分，以及排气通道，其接纳从流动通道穿过弹性体层扩散的气体并将其从弹性体层排出。 装置还可以包括流体连接到流体通道的混合室，以及与限定流动通道的一部分的可偏转膜重叠的控制通道，其中控制通道可操作以改变液体试剂流动的速率 通过流体通道。 所述装置还可包括弹性体层上的刚性塑料层。

公开(公告)号：US08849037B2

公开(公告)日：2014-09-30

申请号：US14071423

申请日：2013-11-04

Applicant: Fluidigm Corporation

Inventor： Simant Dube ,  Gang Sun ,  Lian-She Zhao

IPC: G06K9/36 ,  G06K9/40 ,  G06T7/00 ,  G01N21/64

CPC classification number: G01N21/64 ,  G06T7/0012 ,  G06T7/74 ,  G06T2207/10064 ,  G06T2207/30004

Abstract: A method of processing data associated with fluorescent emissions from a microfluidic device. The method includes performing an auto-focus process associated with a first image of the microfluidic device and performing an auto-exposure process associated with the first image of the microfluidic device. The method also includes capturing a plurality of images of the microfluidic device. The plurality of images are associated with a plurality of thermal cycles. The method further includes performing image analysis of the plurality of captured images to determine a series of optical intensities and performing data analysis of the series of optical intensities to provide a series of change in threshold values.

Abstract translation: 处理与微流体装置的荧光发射有关的数据的方法。 该方法包括执行与微流体装置的第一图像相关联的自动聚焦处理，并且执行与微流体装置的第一图像相关联的自动曝光处理。 该方法还包括捕获微流体装置的多个图像。 多个图像与多个热循环相关联。 该方法还包括执行多个拍摄图像的图像分析以确定一系列光强度并执行该系列光强度的数据分析以提供一系列阈值变化。