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

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Cellular Quiescence

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1686))

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.

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Author information

Authors and 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

    Ayad Eddaoudi & Stephanie Louise Canning

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

    Itaru Kato

Authors
  1. Ayad Eddaoudi
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  2. Stephanie Louise Canning
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  3. Itaru Kato
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Corresponding author

Correspondence to Ayad Eddaoudi .

Editor information

Editors and Affiliations

  1. Texas Children’s Hospital, Baylor College of Medicine, Houston, Texas, USA

    H. Daniel Lacorazza

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Eddaoudi, A., Canning, S.L., Kato, I. (2018). Flow Cytometric Detection of G0 in Live Cells by Hoechst 33342 and Pyronin Y Staining. In: Lacorazza, H. (eds) Cellular Quiescence. Methods in Molecular Biology, vol 1686. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7371-2_3

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  • DOI: https://doi.org/10.1007/978-1-4939-7371-2_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7370-5

  • Online ISBN: 978-1-4939-7371-2

  • eBook Packages: Springer Protocols

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