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。

公开(公告)号：US10221443B2

公开(公告)日：2019-03-05

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 ,  C12Q1/6806 ,  C12M1/00 ,  G01N1/28 ,  C12Q1/6848 ,  B01L3/02 ,  B01L3/00 ,  C12Q1/6841 ,  C12Q1/686

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.

公开(公告)号：US09658142B2

公开(公告)日：2017-05-23

申请号：US15099121

申请日：2016-04-14

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

Inventor： David Richardson ,  Thai Tran ,  Dmitry Derkach ,  Colleen Ziegler

IPC: G01N1/28 ,  B01L3/00

CPC classification number: G01N1/286 ,  B01L3/5027 ,  B01L3/502715 ,  B01L2200/025 ,  B01L2200/0647 ,  B01L2300/0822 ,  B01L2300/0864 ,  B01L2300/0867 ,  B01L2300/0887 ,  B01L2400/0688 ,  G01N2001/2886

Abstract: Methods and apparatuses are provided for extraction and collection of sets of one or more cells from a formalin-fixed paraffin embedded tissue sample. A laser is used to remove sets of one or more cells from an immobilized sample within a microfluidic device, and a fluid flow element washes the removed set of one or more cells away from the immobilized sample, preferably to be collected in a downstream sample collection device. Morphological and/or positional information may be preserved to permit at least one technique (e.g., amplification, sequencing, and/or analytical, optionally performed after cells are lysed downstream of the microfluidic device) to be related to the original spatial location of the set of one or more cells in the immobilized sample.