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Measurement of Cell Death in Mammalian Cells
Author(s) -
Cummings Brian S.,
Schnellmann Rick G.
Publication year - 2021
Publication title -
current protocols
Language(s) - English
Resource type - Journals
ISSN - 2691-1299
DOI - 10.1002/cpz1.210
Subject(s) - programmed cell death , flow cytometry , apoptosis , necrosis , dapi , microbiology and biotechnology , cytotoxicity , viability assay , lactate dehydrogenase , biology , cell , membrane integrity , chemistry , biochemistry , membrane , in vitro , genetics , enzyme
Methods for assessing mammalian cell death are presented in this article, which is divided into six sections: (1) a brief overview of cytotoxicity and pathways of cell death; (2) a method to measure cell death using lactate dehydrogenase (LDH) release as a marker of membrane integrity; (3) a flow cytometry method that simultaneously measures two types of cell death, necrosis and apoptosis; (4) use of fluorescence microscopy and nuclear morphology to assess apoptosis and necrosis; (5) the use of multi‐well plates and high‐content analysis imaging systems to assess nuclear morphology; and (6) a discussion of the use of cytotoxicity assays to determine the mechanisms of cell death. © 2021 Wiley Periodicals LLC. Basic Protocol 1 : Measurement of plasma membrane integrity and viability using LDH release Basic Protocol 2 : Measurement of necrosis and apoptosis using flow cytometry Basic Protocol 3 : Determination of nuclear morphology and membrane integrity Alternate Protocol 1 : Assessment of nuclear morphology and membrane integrity using DAPI and PI Alternate Protocol 2 : Assessment of nuclear morphology using multi‐well plates Basic Protocol 4 : Measurement of time‐dependent toxicity using cell death markers