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

Ayad Eddaoudi; Stephanie Louise Canning; Itaru Kato

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

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

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

Authors

Ayad Eddaoudi¹, Stephanie Louise Canning², Itaru Kato³

Affiliations

¹ Flow Cytometry Core Facility, Camelia Botnar Laboratories, UCL Great Ormond Street Institute of Child Health, 85 Lamb's Conduit, London, WC1N 3JH, UK. a.eddaoudi@ucl.ac.uk.
² Flow Cytometry Core Facility, Camelia Botnar Laboratories, UCL Great Ormond Street Institute of Child Health, 85 Lamb's Conduit, London, WC1N 3JH, UK.
³ Department of Pediatrics, Kyoto University Hospital, 54 Kawaharacho, Syogoin, Sakyu-ku, Kyoto, 606-8507, Japan.

PMID: 29030811
DOI: 10.1007/978-1-4939-7371-2_3

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.

Keywords: Cell cycle; G0; Hoechst 33342; Live-cell sorti ng; Multiparametric flow cytometry; Pyronin Y.

MeSH terms

Benzimidazoles / chemistry*
Flow Cytometry / methods*
Humans
Pyronine / chemistry*
Resting Phase, Cell Cycle*
Staining and Labeling / methods*

Substances

Benzimidazoles
bisbenzimide ethoxide trihydrochloride
Pyronine