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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| Vydáno v: | Acta oncologica Ročník 32; číslo 4; s. 417 - 424 |
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| Médium: | Journal Article Konferenční příspěvek |
| Jazyk: | angličtina |
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Basingstoke
Informa UK Ltd
1993
Taylor & Francis |
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| ISSN: | 0284-186X, 1651-226X |
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| Abstract | 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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| AbstractList | 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. 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.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. |
| Author | Cohen, Gerald M. Sun, Xiao-Ming Ormerod, Michael G. Brown, David Snowden, Roger T. |
| Author_xml | – sequence: 1 givenname: Michael G. surname: Ormerod fullname: Ormerod, Michael G. organization: 1MRC Toxicology Unit. Carshalton, Surrey, England – sequence: 2 givenname: Xiao-Ming surname: Sun fullname: Sun, Xiao-Ming organization: 1MRC Toxicology Unit. Carshalton, Surrey, England – sequence: 3 givenname: David surname: Brown fullname: Brown, David organization: 1MRC Toxicology Unit. Carshalton, Surrey, England – sequence: 4 givenname: Roger T. surname: Snowden fullname: Snowden, Roger T. organization: 1MRC Toxicology Unit. Carshalton, Surrey, England – sequence: 5 givenname: Gerald M. surname: Cohen fullname: Cohen, Gerald M. organization: 1MRC Toxicology Unit. Carshalton, Surrey, England |
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| SubjectTerms | 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 |
| Title | Quantification of Apoptosis and Necrosis by Flow Cytometry |
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