Flow cytometry of apoptotic cell death.
The term apoptosis or programmed cell death defines a genetically encoded cell death program, which is morphologically and biochemically distinct from necrosis or accidental cell death. The characteristic morphological signs of apoptosis (cellular shrinkage, membrane blebbing, nuclear condensation and fragmentation) are the final results of a complex biochemical cascade of events which is an integral part of physiological homeostasis. Techniques designed to identify, quantitate and characterize apoptosis are numerous, but flow cytometry (FCM) remains the methodology of choice to study the apoptotic cascade in relation to cell type, trigger and time. This review outlines the main stages of the apoptotic cascade together with current FCM methods. All FCM apoptosis assays described have a solid experimental basis and have been used successfully in basic research on molecular and biochemical mechanisms of apoptosis. In various clinical settings the ability to follow the apoptotic process in patient samples may offer the rationale for optimal treatment schedules.
Vermes
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Department of Clinical Chemistry, Medical Spectrum Twente, P.O. Box 50000, 7500 KA, Enschede, The Netherlands. i.vermes@wxs.nl
Haanen
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Reutelingsperger
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D017209:Apoptosis / Q000502:physiology*; D002118:Calcium / Q000378:metabolism; D020169:Caspases / Q000032:analysis; D004247:DNA / Q000032:analysis; D004249:DNA Damage; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D019169:Jurkat Cells; D008247:Lysosomes / Q000378:metabolism; D008928:Mitochondria / Q000502:physiology; D010743:Phospholipids / Q000032:analysis; D017494:Proton Pumps / Q000032:analysis; D012313:RNA / Q000032:analysis
D010743:Phospholipids; D017494:Proton Pumps; D012313:RNA; D004247:DNA; D020169:Caspases; D002118:Calcium