Robust high-throughput kinetic analysis of apoptosis with real-time high-content live-cell imaging

Jesse D. Gelles, Jerry Edward Chipuk

Research output: Contribution to journalArticlepeer-review

36 Scopus citations

Abstract

Quantitative and kinetic analyses of apoptotic cell death are integral components of exploring cell biology, measuring cellular stress responses, and performing high-throughput genomic/RNAi/drug screens. Here, we present a detailed method that integrates robust kinetic real-time high-content imaging with Annexin V labelling to provide a highly sensitive, accurate, simple and zero-handling approach to quantify extrinsic and intrinsic inducers of apoptosis. The sensitivity of this non-toxic method outperforms previous high-throughput methodologies using viability dyes or caspase-activation reporters. This method also incorporates a multiplex adaptation to integrate variability in cell number due to treatment-induced proliferation changes and the detachment of dying cells. Compared to Annexin V detection by flow cytometry, this method is 10-fold more sensitive, eliminates extensive sample handling and processing, and provides real-time kinetics of apoptosis at both single-cell and population-level resolutions.

Original languageEnglish
Article numbere2493
JournalCell Death and Disease
Volume7
Issue number12
DOIs
StatePublished - 1 Dec 2016

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