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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.<\/jats:p>","DOI":"10.1038\/cddis.2016.332","type":"journal-article","created":{"date-parts":[[2016,12,1]],"date-time":"2016-12-01T08:47:52Z","timestamp":1480582072000},"page":"e2493-e2493","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":63,"title":["Robust high-throughput kinetic analysis of apoptosis with real-time high-content live-cell imaging"],"prefix":"10.1038","volume":"7","author":[{"given":"Jesse D","family":"Gelles","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jerry Edward","family":"Chipuk","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2016,12,1]]},"reference":[{"key":"BFcddis2016332_CR1","doi-asserted-by":"crossref","first-page":"1415","DOI":"10.1182\/blood.V84.5.1415.bloodjournal8451415","volume":"84","author":"G Koopman","year":"1994","unstructured":"Koopman G, Reutelingsperger C, Kuijten G, Keehnen R, Pals S, van Oers M . 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