title: Measuring Apoptosis by Microscopy and Flow Cytometry.
abstract: Apoptosis is a mode of programmed cell death that plays an important role during development and in the maintenance of tissue homeostasis. Numerous physiological as well as pathological stimuli trigger apoptosis such as engagement of Fas, TRAIL, or TNF receptors, growth factor deprivation, hypoxia, or exposure to cytotoxic drugs. Apoptosis is coordinated from within by members of the caspase family of cysteine proteases that, upon activation, trigger a series of morphological changes including cell shrinkage, extensive plasma membrane blebbing, chromatin condensation, DNA hydrolysis, and nuclear fragmentation. These dramatic structural and biochemical alterations result not only in the controlled dismantling of the cell, but also in the efficient recognition and removal of apoptotic cells by phagocytes. Necrosis, which is typically nonprogrammed or imposed upon the cell by overwhelming membrane or organelle damage, is characterized by rapid plasma membrane rupture followed by organelle and cell swelling. Necrosis is often provoked by infectious agents or a severe departure from physiological conditions. This unit describes protocols for the measurement of apoptosis and for distinguishing apoptosis from necrosis.
authors.affiliation: Molecular Cell Biology Laboratory, Department of Genetics, Trinity College, Dublin, Ireland.
authors.forename: Emilie
authors.initials: E
authors.lastname: Hollville
authors.affiliation: Molecular Cell Biology Laboratory, Department of Genetics, Trinity College, Dublin, Ireland.
authors.forename: Seamus J
authors.initials: SJ
authors.lastname: Martin
mesh_terms: D017304:Annexin A5; D017209:Apoptosis; D020169:Caspases / Q000378:metabolism; D002462:Cell Membrane; D004396:Coloring Agents; D053938:DNA Fragmentation; D004801:Eosine Yellowish-(YS); D005434:Flow Cytometry / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D020287:In Situ Nick-End Labeling; D007202:Indicators and Reagents; D019169:Jurkat Cells; D008751:Methylene Blue; D008853:Microscopy / Q000379:methods*; D011419:Propidium
chemical_list: D017304:Annexin A5; D004396:Coloring Agents; D007202:Indicators and Reagents; D011419:Propidium; D020169:Caspases; D008751:Methylene Blue; D004801:Eosine Yellowish-(YS)
keywords: apoptosis; cell morphology; flow cytometry; microscopy; necrosis
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