Abstract
The apoptotic response to various stimuli is an important part of immune regulation, and the ability to identify apoptotic lymphocytes within a complex population is a prerequisite to a more detailed understanding of its role in vivo, We described a flow cytometric technique which utilizes viable cells and enables simultaneous identification of apoptotic cells and analyses of immunophenotype, cell cycle progression, membrane integrity and light scatter properties. It is based upon analysis of two regions of the emission spectrum of the DNA-binding vital dye hoechst 33342. We established a precise correlation between the ratio of red to blue fluorescence emission and apoptosis based upon nuclear morphology and the presence of characteristic DNA degradation patterns. In human peripheral blood lymphocytes (PBL) and mouse thymocytes we incorporated light scatter properties, cell cycle stage, relevant cell surface immunophenotypic markers (CD25 or CD4) and CD8) and a marker of plasma membrane integrity (merocyanine 540) to enable multiparametric phenotyping of apoptotic cells. We show that staurosporine-induced apoptosis of ConA-stimulated PBL is not correlated with cell cycle stage but is selective for activated cells since the fre...Continue Reading
References
Jan 1, 1976·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·S A Latt, G Stetten
Jan 1, 1992·Cytometry·Z DarzynkiewiczF Traganos
Sep 18, 1992·Journal of Immunological Methods·J A HardinP A Lopez
Jan 1, 1992·Cytometry·A B LyonsA H Maddy
Jan 1, 1990·Cytometry·J W Ellwart, P Dörmer
Feb 3, 1992·Biochimica Et Biophysica Acta·C DiveA H Wyllie
Mar 1, 1991·Journal of Immunological Methods·W SwatP Kisielow
Jul 1, 1985·Cytometry·J V WatsonP J Smith
Mar 1, 1986·Journal of Cellular Physiology·B J Del BuonoR A Schlegel
Apr 1, 1985·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·T Stokke, H B Steen
Apr 10, 1980·Nature·A H Wyllie
Jul 27, 1983·Biochimica Et Biophysica Acta·P WilliamsonR A Schlegel
Jan 1, 1994·Cytometry·I SchmidJ V Giorgi
Sep 1, 1994·Cytometry·F BellocF Lacombe
Jul 17, 1995·Journal of Immunological Methods·I VermesC Reutelingsperger
Dec 1, 1994·Immunology Today·G NúñezM González-García
Nov 1, 1993·The Journal of Experimental Medicine·S AndjelićJ Nikolić-Zugić
Nov 1, 1993·Immunology Today·A N AkbarG Janossy
Aug 1, 1993·The Journal of Experimental Medicine·A N AkbarG Janossy
Jan 1, 1993·Cytometry·M G OrmerodG M Cohen
Feb 23, 1993·Biochimica Et Biophysica Acta·W StillwellL J Jenski
Mar 1, 1993·Immunology Today·J J Cohen
Citations
Jun 10, 1996·Journal of Immunological Methods·S ReidE C Snow
Mar 28, 1997·Journal of Immunological Methods·J SindermannG Breithardt
Aug 6, 1998·Journal of Immunological Methods·C E AllsoppJ Langhorne
Oct 1, 1996·Molecular Immunology·S Reid, E C Snow
Sep 25, 1999·Human Gene Therapy·K E PollokD A Williams
May 6, 2006·Journal of Biomedical Optics·Yinghua SunLaura Marcu
Jan 28, 2006·BMC Cell Biology·Francesco FabbriMarina Marini
Dec 14, 2001·Biochemical and Biophysical Research Communications·Z ShifferM Deutsch
Jul 6, 2010·Microbial Pathogenesis·Lucie ZivnaJiri Stulik
Aug 4, 2004·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Timothy W PetersenGer van den Engh
May 8, 2004·Biochemical Pharmacology·Masaru SaitohTomoyuki Nishizaki
Jan 10, 2002·Redox Report : Communications in Free Radical Research·M G Ormerod
Sep 20, 2002·The Journal of Toxicological Sciences·Kazuyuki NagamiHiroyoshi Matsumoto
May 9, 2000·Memórias do Instituto Oswaldo Cruz·A L BerthoS G Coutinho
Dec 20, 1999·Cytometry·C MatteucciA Mastino
Aug 17, 2006·Blood·Jorg A KrugerRalph A Reisfeld
Jul 9, 1999·Journal of Biomedical Materials Research·T FabreA Durandeau
Aug 30, 2003·Archives of Pathology & Laboratory Medicine·Xinbo Zhang, Frederick L Kiechle
Oct 25, 2003·Brain Research·Masaru SaitohTomoyuki Nishizaki
Jan 11, 2001·Archives of Pathology & Laboratory Medicine·X Zhang, F L Kiechle