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

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

SUBMITTER: Gelles JD 

PROVIDER: S-EPMC5261025 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature