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激光捕获显微切割:从基因组到染色体,从复杂组织到单细胞分析。

Laser Capture Microdissection: From Genomes to Chromosomes, from Complex Tissue to Single-Cell Analysis.

机构信息

Department of Plant Developmental Genetics, Institute of Biophysics of the Czech Academy of Sciences, Brno, Czech Republic.

出版信息

Methods Mol Biol. 2023;2672:163-175. doi: 10.1007/978-1-0716-3226-0_9.

Abstract

Laser microdissection (LM) is a powerful tool for various molecular analyses providing pure samples for genomic, transcriptomic, and proteomic studies. Cell subgroups, individual cells, or even chromosomes can be separated via laser beam from complex tissues, visualized under the microscope, and used for subsequent molecular analyses. This technique provides information on nucleic acids and proteins, keeping their spatiotemporal information intact. In short, the slide with tissue is placed under the microscope, imaged by a camera onto a computer screen, where the operator selects cells/chromosomes based on morphology or staining and commands the laser beam to cut the specimen following the selected path. Samples are then collected in a tube and subjected to downstream molecular analysis, such as RT-PCR, next-generation sequencing, or immunoassay.

摘要

激光显微切割(LM)是一种强大的工具,可用于各种分子分析,为基因组、转录组和蛋白质组学研究提供纯样品。通过激光束,可以从复杂的组织中分离出细胞亚群、单个细胞甚至染色体,在显微镜下观察,并用于随后的分子分析。该技术提供了关于核酸和蛋白质的信息,同时保持其时空信息的完整性。简而言之,将载有组织的载玻片置于显微镜下,通过相机将组织图像成像到计算机屏幕上,操作人员根据形态或染色选择细胞/染色体,并命令激光束沿着选定的路径切割标本。然后将样品收集在管中,并进行下游分子分析,如 RT-PCR、下一代测序或免疫测定。

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