Optimal detection of apoptosis by flow cytometry depends on cell morphology.
Flow cytometry has recently become a choice technique for the quantitative analysis of apoptosis. Monoparametric DNA analysis usually allows identification of apoptotic cells as a "subdiploid" peak. Progression through apoptosis leads to chromatin condensation, nuclear fragmentation and eventually to cell disruption. Thus, a major problem for the flow cytometric analysis of apoptotic populations is discrimination between debris and apoptotic cells. Here we demonstrate that the best parameter on which to make such a distinction is the DNA content, no matter what type of cell is studied. In contrast, discrimination between apoptotic, non-apoptotic cells, and debris is possible on the basis of scattering signals only in few selected cases, depending on the morphology of the intact cells.
Zamai
L
L
Laboratorio Biologia Cellulare e Microscopia Elettronica, Istituto Ortopedico Rizzoli, Bologna, Italy.
Falcieri
E
E
Zauli
G
G
Cataldi
A
A
Vitale
M
M
D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D002843:Chromatin / Q000737:chemistry / Q000648:ultrastructure; D004247:DNA / Q000032:analysis; D004273:DNA, Neoplasm / Q000032:analysis; D005434:Flow Cytometry / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000166:cytology* / Q000648:ultrastructure; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000235:genetics / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D013950:Thymus Gland / Q000737:chemistry / Q000166:cytology* / Q000648:ultrastructure; D014407:Tumor Cells, Cultured
D002843:Chromatin; D004273:DNA, Neoplasm; D004247:DNA