STAR Protocols (Sep 2022)

Coupling live-cell imaging and in situ isolation of the same single cell to profile the transient states of predicted drug-tolerant cells

  • Benjamin Bian,
  • Agnès Paquet,
  • Marie-Jeanne Arguel,
  • Mickael Meyer,
  • Ludovic Peyre,
  • Asma Chalabi,
  • Marielle Péré,
  • Kevin Lebrigand,
  • Rainer Waldmann,
  • Pascal Barbry,
  • Paul Hofman,
  • Jérémie Roux

Journal volume & issue
Vol. 3, no. 3
p. 101600

Abstract

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Summary: Cell response variability is a starting point in cancer drug resistance that has been difficult to analyze because the tolerant cell states are short lived. Here, we present fate-seq, an approach to isolate single cells in their transient states of drug sensitivity or tolerance before profiling. The drug response is predicted in live cells, which are laser-captured by microdissection before any drug-induced change can alter their states. This framework enables the identification of the cell-state signatures causing differential cell decisions upon treatment.For complete details on the use and execution of this protocol, please refer to Meyer et al. (2020). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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