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    SYSTEM AND METHOD FOR LASER LYSIS
    4.
    发明申请
    SYSTEM AND METHOD FOR LASER LYSIS 审中-公开
    激光雷达系统和方法

    公开(公告)号：US20160237476A1

    公开(公告)日：2016-08-18

    申请号：US15135150

    申请日：2016-04-21

    Applicant: ARIZONA BOARD OF REGENTS ON BEHALF OF ARIZONA STATE UNIVERSITY

    Inventor： Deirdre Meldrum ,  Shih-Hui (Joseph) Chao ,  Thai Tran ,  Laimonas Kelbauskas ,  Jeff Houkal ,  Andrew Hatch ,  Weimin Gao ,  David Richardson

    IPC: C12Q1/68

    CPC classification number: C12Q1/6806 ,  B01L3/0293 ,  B01L3/502761 ,  B01L2200/0668 ,  B01L2300/0816 ,  B01L2300/0829 ,  B01L2300/0864 ,  C12M47/06 ,  C12Q1/6841 ,  C12Q1/6848 ,  C12Q1/686 ,  G01N1/286 ,  G01N2001/2886 ,  C12Q2547/101

    Abstract: Systems and methods for in situ laser lysis for analysis of biological tissue (live, fixed, frozen or otherwise preserved) at single cell resolution in 3D. For example, a system and method for lysing individual cells in situ, including the steps of capturing a tissue sample comprising a cellular content, subjecting the tissue sample to a stream of continuous fluid flow, lysing a selected area of the tissue sample with a laser, thereby releasing at least a portion of the cellular content from the tissue sample, recovering at least one target molecule from the cellular content in the stream, and processing at least one target molecule is provided. The system collects cellular contents, performs highly multiplexed (RT-qPCR or RNA-seq), and sequentially (cell-by-cell) reconstructs a 3D spatial map of mRNA expression of the tissue with a large number of genes. A 3D spatial map of the DNA, RNA, and/or proteins can be generated for each cell in the tissue.

    Abstract translation: 用于原位激光裂解的系统和方法，用于分析3D中单细胞分辨率的生物组织（活，固定，冷冻或以其他方式保存）。 例如，用于原位裂解单个细胞的系统和方法，包括以下步骤：捕获包含细胞内容物的组织样品，使组织样品经受连续流体流的流动，用激光裂解组织样品的选定区域 从而从组织样品释放至少一部分细胞含量，从流中的细胞含量回收至少一种靶分子，并且提供至少一种靶分子的加工。 系统收集细胞内容，进行高度多重化（RT-qPCR或RNA-seq），并依次（逐个细胞）重构具有大量基因的组织的mRNA表达的3D空间图。 可以为组织中的每个细胞产生DNA，RNA和/或蛋白质的3D空间图。

    Systems and methods for continuous flow digital droplet polymerase chain reaction bioanalysis
    5.
    发明授权
    Systems and methods for continuous flow digital droplet polymerase chain reaction bioanalysis 有权

    公开(公告)号：US11618030B2

    公开(公告)日：2023-04-04

    申请号：US17389253

    申请日：2021-07-29

    Applicant: Arizona Board of Regents on Behalf of Arizona State University

    Inventor： Cody Youngbull ,  Andrew Hatch ,  Tathagata Ray ,  Andrew Larsen ,  Matthew Underhill

    IPC: B01L7/00 ,  G01N35/10 ,  B01L3/00 ,  C12Q1/686 ,  G01N1/10 ,  G01N35/00 ,  B01F33/302

    Abstract: Systems and methods for continuous flow polymerase chain reaction (PCR) are provided. The system comprises an injector, a mixer, a coalescer, a droplet generator, a detector, a digital PCR system, and a controller. The injector takes in a sample, partitions the sample into sample aliquots with the help of an immiscible oil phase, dispenses waste, and sends the sample aliquot to the mixer. The mixer mixes the sample aliquot with a PCR master mix and diluting water, dispenses waste, and sends the sample mixture (separated by an immiscible oil) to the coalescer. The coalescer coalesces the sample mixture with primers dispensed from a cassette, dispenses waste, and sends the reaction mixture (separated by an immiscible oil) to the droplet generator. The droplet generator converts the sample mixture into an emulsion where aqueous droplets of the reaction mixture are maintained inside of an immiscible oil phase and dispenses droplets to the digital PCR system. The digital PCR system amplifies target DNAs in the droplets. The detector detects target DNAs in the droplets. The controller controls the system to run automatically and continuously.

    Systems and methods for continuous flow digital droplet polymerase chain reaction bioanalysis
    7.
    发明授权
    Systems and methods for continuous flow digital droplet polymerase chain reaction bioanalysis 有权

    公开(公告)号：US11123740B2

    公开(公告)日：2021-09-21

    申请号：US15739318

    申请日：2016-06-29

    Applicant: ARIZONA BOARD OF REGENTS ON BEHALF OF ARIZONA STATE UNIVERSITY

    Inventor： Cody Youngbull ,  Andrew Hatch ,  Tathagata Ray ,  Andrew Larsen ,  Matthew Underhill

    IPC: B01L7/00 ,  G01N35/10 ,  B01L3/00 ,  C12Q1/686 ,  G01N1/10 ,  G01N35/00 ,  B01F13/00

    Abstract: Systems and methods for continuous flow polymerase chain reaction (PCR) are provided. The system comprises an injector, a mixer, a coalescer, a droplet generator, a detector, a digital PCR system, and a controller. The injector takes in a sample, partitions the sample into sample aliquots with the help of an immiscible oil phase, dispenses waste, and sends the sample aliquot to the mixer. The mixer mixes the sample aliquot with a PCR master mix and diluting water, dispenses waste, and sends the sample mixture (separated by an immiscible oil) to the coalescer. The coalescer coalesces the sample mixture with primers dispensed from a cassette, dispenses waste, and sends the reaction mixture (separated by an immiscible oil) to the droplet generator. The droplet generator converts the sample mixture into an emulsion where aqueous droplets of the reaction mixture are maintained inside of an immiscible oil phase and dispenses droplets to the digital PCR system. The digital PCR system amplifies target DNAs in the droplets. The detector detects target DNAs in the droplets. The controller controls the system to run automatically and continuously.

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