Flow cytometry of apoptosis.
Common methods applicable to flow cytometry make it possible to: (1) identify and quantify dead or dying cells, (2) reveal a mode of cell death (apoptosis or necrosis), and (3) study mechanisms involved in cell death. Gross changes in cell morphology and chromatin condensation, which occur during apoptosis, can be detected by analysis with laser light beam scattering. Early events of apoptosis, dissipation of the mitochondrial transmembrane potential and caspase activation, can be detected using either fluorochrome reporter groups or appropriate antibodies. Exposure of phosphatidylserine on the exterior surface of the plasma membrane can be detected by the binding of fluoresceinated annexin V. Another apoptotic event, DNA fragmentation based on DNA content of cells with fractional ("sub-G1") or DNA strand-break labeling, TUNEL; or In Situ End Labeling, ISEL;. Still another hallmark of apoptosis is the activation of tissue transglutaminase (TGase), the enzyme that crosslinks protein and thereby makes them less immunogenic. The major advantage of flow cytometry in these applications is that it provides the possibility of multiparametric measurements of cell attributes.
Pozarowski
Piotr
P
School of Medicine, Lublin, Poland.
Grabarek
Jerzy
J
Darzynkiewicz
Zbigniew
Z
D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects; D002103:Cadaverine; D002232:Carbocyanines / Q000378:metabolism; D020169:Caspases / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D053938:DNA Fragmentation / Q000187:drug effects; D003902:Detergents / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism; D018922:HL-60 Cells; D006801:Humans; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D007211:Indoles; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008856:Microscopy, Fluorescence; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D011419:Propidium / Q000378:metabolism; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D020112:Rhodamine 123 / Q000378:metabolism; D011503:Transglutaminases / Q000378:metabolism
D017304:Annexin A5; D002232:Carbocyanines; D003902:Detergents; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D007211:Indoles; D020112:Rhodamine 123; D011419:Propidium; C007293:DAPI; C007392:3,3'-dihexyl-2,2'-oxacarbocyanine; D004247:DNA; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D011065:Poly(ADP-ribose) Polymerases; D020169:Caspases; D019204:GTP-Binding Proteins; D002103:Cadaverine