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[荧光激活细胞分选后mRNA定量分析的发展]

[Development of mRNA quantification after fluorescence activated cell sorting].

作者信息

Yamada Hiroya

机构信息

Department of Hygiene, Fujita Health University School of Medicine, Toyoake 470-1192, Japan.

出版信息

Rinsho Byori. 2012 Feb;60(2):139-44.

Abstract

Recent studies using stem cells or cancer stem cells have revealed the importance of detecting minor populations of cells in blood or tissue and analyzing their biological characteristics. These cells can be separated from other cells by fluorescence-activated cell sorting (FACS) using antibodies that bind to surface antigen; however, the majority of these cells do not have specific surface antigens. Furthermore, the cells must be kept alive throughout the procedure to analyze their biological characteristics. To improve these limitations, we have established a novel laboratory test named mRNA quantification after fluorescence-activated cell sorting(FACS-mQ). Using this method, cells are sorted by a specific gene expression pattern, and the gene expression profile in the sorted cells can be easily analyzed. In this feature, we will introduce the typical applications of FACS-mQ targeting intracellular mRNA and intracellular nuclear antigens. These methods will hopefully contribute to the accumulation of knowledge regarding human stem cells, cancer stem cells, and small populations of cells, the biological characteristics of which are mostly unknown.

摘要

最近使用干细胞或癌症干细胞的研究揭示了检测血液或组织中少量细胞群体并分析其生物学特性的重要性。这些细胞可以使用与表面抗原结合的抗体通过荧光激活细胞分选(FACS)从其他细胞中分离出来;然而,这些细胞中的大多数没有特异性表面抗原。此外,在整个过程中必须保持细胞存活以分析其生物学特性。为了改善这些局限性,我们建立了一种名为荧光激活细胞分选后mRNA定量(FACS-mQ)的新型实验室检测方法。使用这种方法,细胞通过特定的基因表达模式进行分选,并且可以轻松分析分选细胞中的基因表达谱。在本专题中,我们将介绍针对细胞内mRNA和细胞核内抗原的FACS-mQ的典型应用。这些方法有望有助于积累有关人类干细胞、癌症干细胞和少量细胞群体的知识,而这些细胞群体的生物学特性大多未知。

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