PMID: 8613668Feb 5, 1996Paper

Flow cytometric ratio analysis of the Hoechst 33342 emission spectrum: multiparametric characterization of apoptotic lymphocytes

Journal of Immunological Methods
L ChiuJ F Dunne

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

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Citations

May 6, 2006·Journal of Biomedical Optics·Yinghua SunLaura Marcu
Dec 14, 2001·Biochemical and Biophysical Research Communications·Z ShifferM Deutsch
Aug 4, 2004·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Timothy W PetersenGer van den Engh
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
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

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