Quantification of Apoptosis and Necrosis by Flow Cytometry

Apoptosis and necrosis are two important mechanisms of cell death. Several methods have recently been described for quantifying apoptotic cells by flow cytometry. We report a novel method for the quantification and separation of viable normal and apoptotic cells. We have applied this method both to...

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Bibliographic Details
Published in:Acta oncologica Vol. 32; no. 4; pp. 417 - 424
Main Authors: Ormerod, Michael G., Sun, Xiao-Ming, Brown, David, Snowden, Roger T., Cohen, Gerald M.
Format: Journal Article Conference Proceeding
Language:English
Published: Basingstoke Informa UK Ltd 1993
Taylor & Francis
Subjects:
Ageing, cell death
Animals
Apoptosis - physiology
Benzimidazoles - pharmacokinetics
Biological and medical sciences
Cell Death - physiology
Cell Membrane - metabolism
Cell physiology
DNA - analysis
Flow Cytometry
Fluorescent Dyes - pharmacokinetics
Fundamental and applied biological sciences. Psychology
Male
Mice
Molecular and cellular biology
Necrosis - pathology
Propidium - pharmacokinetics
Rats
Rats, Inbred F344
Thymus Gland - chemistry
Thymus Gland - cytology
Trypan Blue - pharmacokinetics
Flow cytometry
Vertebrata
Mammalia
Rat
Mouse
Cell death
Thymocyte
Animal
Rodentia
Hematopoietic cell
Quantization
Necrosis
ISSN:0284-186X, 1651-226X
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Summary:Apoptosis and necrosis are two important mechanisms of cell death. Several methods have recently been described for quantifying apoptotic cells by flow cytometry. We report a novel method for the quantification and separation of viable normal and apoptotic cells. We have applied this method both to immature rat thymocytes treated with a variety of agents and to a murine haemopoetic cell line after withdrawal of a growth factor. The cells were incubated with two dyes which give fluorescent complexes when bound to DNA, the bis-benzimidazole, Hoechst 33342, and propidium iodide. Three populations were identified and characterized. On excitation with UV radiation, dead cells fluoresced red due to the uptake of propidium iodide whereas apoptotic cells fluoresced bright blue; normal cells showed low blue, low red fluorescence. In this paper, we demonstrate how this method may be used to help to distinguish between cell death by apoptosis and necrosis.
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ISSN:0284-186X
1651-226X
DOI:10.3109/02841869309093620