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Cell-Based High Content Analysis of Cell Proliferation and Apoptosis

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High Content Screening

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1683))

Abstract

High content imaging-based cell cycle analysis allows multiplexing of various parameters including DNA content, DNA synthesis, cell proliferation, and other cell cycle markers such as phosho-histone H3. 5′-Ethynyl-2′-deoxyuridine (EdU) incorporation is a thymidine analog that provides a sensitive method for the detection of DNA synthesis in proliferating cells that is a more convenient method than the traditional BrdU detection by antibody. Caspase 3 is activated in programmed cell death induced by both intrinsic (mitochondrial) and extrinsic factors (death ligand). Cell cycle and apoptosis are common parameters studied in the phenotypic analysis of compound toxicity and anti-cancer drugs. In this chapter, we describe methods for the detection of s-phase cell cycle progression by EdU incorporation, and caspase 3 activation using the CellEvent caspase 3/7 detection reagent.

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Authors and Affiliations

  1. Thermo Fisher Scientific, 29851 Willow Creek Road, Eugene, OR, 97402, USA

    Bhaskar S. Mandavilli, Michelle Yan & Scott Clarke

Authors
  1. Bhaskar S. Mandavilli
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  2. Michelle Yan
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  3. Scott Clarke
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Corresponding author

Correspondence to Bhaskar S. Mandavilli .

Editor information

Editors and Affiliations

  1. Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, Pennsylvania, USA

    Paul A. Johnston

  2. Bahama Bio, LLC, Bahama, North Carolina, USA

    Oscar J. Trask

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Mandavilli, B.S., Yan, M., Clarke, S. (2018). Cell-Based High Content Analysis of Cell Proliferation and Apoptosis. In: Johnston, P., Trask, O. (eds) High Content Screening. Methods in Molecular Biology, vol 1683. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7357-6_4

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  • DOI: https://doi.org/10.1007/978-1-4939-7357-6_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7355-2

  • Online ISBN: 978-1-4939-7357-6

  • eBook Packages: Springer Protocols

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