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Cell Cycle Mapping Using Multiplexed Immunofluorescence

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Cell Cycle Control

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2740))

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Abstract

The development of technologies that allow measurement of the cell cycle at the single-cell level has revealed novel insights into the mechanisms that regulate cell cycle commitment and progression through DNA replication and cell division. These studies have also provided evidence of heterogeneity in cell cycle regulation among individual cells, even within a genetically identical population. Cell cycle mapping combines highly multiplexed imaging with manifold learning to visualize the diversity of “paths” that cells can take through the proliferative cell cycle or into various states of cell cycle arrest. In this chapter, we describe a general protocol of the experimental and computational components of cell cycle mapping. We also provide a comprehensive guide for the design and analysis of experiments, discussing key considerations in detail (e.g., antibody library preparation, analysis strategies, etc.) that may vary depending on the research question being addressed.

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Authors and Affiliations

  1. Department of Cell Biology, Center for Biologic Imaging (CBI), University of Pittsburgh, Pittsburgh, PA, USA

    Katarzyna M. Kedziora

  2. Department of Computational and Systems Biology, UPMC Hillman Cancer Center, University of Pittsburgh, Pittsburgh, PA, USA

    Wayne Stallaert

Authors
  1. Katarzyna M. Kedziora
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  2. Wayne Stallaert
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Correspondence to Wayne Stallaert .

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Editors and Affiliations

  1. Université de Montpellier, Centre de Recherche en Biologie cellulaire de Montpellier (CRBM), CNRS UMR 5237, Montpellier Cedex 5, France

    Anna Castro

  2. Université de Montpellier, Centre de Recherche en Biologie cellulaire de Montpellier (CRBM), CNRS UMR 5237, Montpellier Cedex 5, France

    Benjamin Lacroix

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Kedziora, K.M., Stallaert, W. (2024). Cell Cycle Mapping Using Multiplexed Immunofluorescence. In: Castro, A., Lacroix, B. (eds) Cell Cycle Control. Methods in Molecular Biology, vol 2740. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3557-5_15

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  • DOI: https://doi.org/10.1007/978-1-0716-3557-5_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3556-8

  • Online ISBN: 978-1-0716-3557-5

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