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用于人类大脑基因表达谱分析的细胞类型特异性激光捕获显微切割技术

Cell Type-Specific Laser Capture Microdissection for Gene Expression Profiling in the Human Brain.

作者信息

Mauney Sarah A, Woo Tsung-Ung W, Sonntag Kai C

机构信息

Laboratory of Cellular Neuropathology, McLean Hospital, Harvard Medical School, Belmont, MA, USA.

Basic Neuroscience Division, Department of Psychiatry, McLean Hospital, Harvard Medical School, Belmont, MA, USA.

出版信息

Methods Mol Biol. 2018;1723:203-221. doi: 10.1007/978-1-4939-7558-7_11.

Abstract

Cell type-specific laser microdissection technologies in combination with molecular techniques to determine gene expression profiles have become powerful tools to gain insight into the neurobiological basis of neural circuit disturbances in various neurologic or psychiatric diseases. To identify specific cell populations in human postmortem brain tissue, one can use the inherent properties of the cells, such as pigmentation and morphology or their structural composition through immunohistochemistry (IHC). Here, we describe the isolation of homogeneous neurons and oligodendrocytes and the extraction of high-quality RNA from these cells in human postmortem brain using a combination of rapid IHC, Nissl staining, or simple morphology with Laser Capture Microdissection (LCM), or Laser Microdissection (LMD).

摘要

将细胞类型特异性激光显微切割技术与分子技术相结合以确定基因表达谱,已成为深入了解各种神经或精神疾病中神经回路紊乱的神经生物学基础的有力工具。为了在人类尸检脑组织中识别特定的细胞群体,可以利用细胞的固有特性,如色素沉着、形态或通过免疫组织化学(IHC)检测其结构组成。在这里,我们描述了使用快速免疫组织化学、尼氏染色或简单形态学与激光捕获显微切割(LCM)或激光显微切割(LMD)相结合的方法,从人类尸检脑组织中分离出均匀的神经元和少突胶质细胞,并从这些细胞中提取高质量的RNA。

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