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Titolo:
Flow cytometric analysis of apoptotic subpopulations with a combination ofannexin V-FITC, propidium iodide, and SYTO 17
Autore:
Eray, M; Matto, M; Kaartinen, M; Andersson, LC; Pelkonen, J;
Indirizzi:
Univ Helsinki, Haartman Inst, Dept Pathol, FIN-00014 Helsinki, Finland Univ Helsinki Helsinki Finland FIN-00014 ol, FIN-00014 Helsinki, Finland Univ Helsinki Hosp, Helsinki, Finland Univ Helsinki Hosp Helsinki Finland iv Helsinki Hosp, Helsinki, Finland Univ Helsinki, Haartman Inst, Dept Bacteriol & Immunol, FIN-00014 Helsinki, Finland Univ Helsinki Helsinki Finland FIN-00014 ol, FIN-00014 Helsinki, Finland Univ Kuopio, Dept Clin Microbiol, FIN-70211 Kuopio, Finland Univ Kuopio Kuopio Finland FIN-70211 icrobiol, FIN-70211 Kuopio, Finland Kuopio Univ Hosp, Dept Pediat, SF-70210 Kuopio, Finland Kuopio Univ Hosp Kuopio Finland SF-70210 ediat, SF-70210 Kuopio, Finland
Titolo Testata:
CYTOMETRY
fascicolo: 2, volume: 43, anno: 2001,
pagine: 134 - 142
SICI:
0196-4763(20010201)43:2<134:FCAOAS>2.0.ZU;2-J
Fonte:
ISI
Lingua:
ENG
Soggetto:
PROGRAMMED CELL-DEATH; B-CELLS; PHOSPHATIDYLSERINE EXPRESSION; LIVE CELLS; CD40; NECROSIS; ANTIGEN; ASSAY; ACTIVATION; INHIBITION;
Keywords:
apoptosis; B cells; annexin V-FITC; PI; SYTO 17;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
38
Recensione:
Indirizzi per estratti:
Indirizzo: Eray, M Univ Helsinki, Haartman Inst, Dept Pathol, POB 21, FIN-00014 Helsinki, Finland Univ Helsinki POB 21 Helsinki Finland FIN-00014 Helsinki, Finland
Citazione:
M. Eray et al., "Flow cytometric analysis of apoptotic subpopulations with a combination ofannexin V-FITC, propidium iodide, and SYTO 17", CYTOMETRY, 43(2), 2001, pp. 134-142

Abstract

Background: We have previously characterized apoptotic cell death induced in a follicular lymphoma cell line, HF-1, after triggering via the B-cell receptor (BCR) or treatment with Ca2+ Ionophore A23187. We analyzed the kinetics of apoptosis induced by these two treatments, as two alternative models of classical apoptosis, by flow cytometry using a novel combination of cytofluorometric stains. Methods: Cells were stained with a combination of Annexin V-FITC, propidium iodide (PI), and SYTO 17 and analyzed by a two-laser flow cytometry system using 488-nm argon and 633-nm HeNe air-cooled lasers. Results: In both apoptotic models, the first apoptotic cells were detectedby SYTO 17 staining. The alteration in SYTO 17 staining intensity was followed by an increased uptake of PI. Finally, the apoptotic cells were labeled with Annexin V in BCR-induced apoptosis. On the contrary, on treatment with Ca2+ Ionophore A23187, cells became positive for Annexin V earlier than for PI. Conclusions: The novel cytofluorometric dye, SYTO 17, discriminates apoptotic alterations before Annexin V and PI. PI also discriminates apoptotic alterations before the loss of plasma membrane asymmetry by BCR but not by Ca2+ Ionophore A23187-induced apoptosis. Finally, the combination of these three cytofluorometric dyes allows effective detection of apoptotic subpopulations and ordering of apoptotic events by flow cytometry. (C) 2001 Wiley-Liss. Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 04/07/20 alle ore 15:00:35