通过Hoechst 33342和派洛宁Y染色对活细胞中G0期进行流式细胞术检测

Flow Cytometric Detection of G0 in Live Cells by Hoechst 33342 and Pyronin Y Staining.

作者信息

Eddaoudi Ayad, Canning Stephanie Louise, Kato Itaru

机构信息

Flow Cytometry Core Facility, Camelia Botnar Laboratories, UCL Great Ormond Street Institute of Child Health, 85 Lamb's Conduit, London, WC1N 3JH, UK.

Department of Pediatrics, Kyoto University Hospital, 54 Kawaharacho, Syogoin, Sakyu-ku, Kyoto, 606-8507, Japan.

出版信息

Methods Mol Biol. 2018;1686:49-57. doi: 10.1007/978-1-4939-7371-2_3.

DOI:10.1007/978-1-4939-7371-2_3

PMID:29030811

Abstract

Hoechst 33342 and Pyronin Y double staining can be used to measure DNA and RNA content in live cells by flow cytometry. Quiescent cells at G0 phase have the same amount of DNA as cells at G1 phase but lower RNA levels compared to proliferating cells. Therefore, resting cells in G0 phase can be distinguished from proliferating cells in G1, S, and G2 M phases. This chapter describes a protocol for double staining of live cells with Hoechst 33342 and Pyronin Y. Combined with immunophenotyping of intact and live cells Hoechst 33342 and Pyronin Y staining is a powerful noninvasive method for the analysis and isolation of quiescent cells from any defined cell population.

摘要

Hoechst 33342和派洛宁Y双重染色可用于通过流式细胞术测量活细胞中的DNA和RNA含量。处于G0期的静止细胞与G1期细胞的DNA含量相同，但与增殖细胞相比，RNA水平较低。因此，G0期的静止细胞可以与G1、S和G2 M期的增殖细胞区分开来。本章介绍了用Hoechst 33342和派洛宁Y对活细胞进行双重染色的方法。结合完整活细胞的免疫表型分析，Hoechst 33342和派洛宁Y染色是一种从任何定义的细胞群体中分析和分离静止细胞的强大非侵入性方法。