PMID: 11309809Apr 20, 2001Paper

Supravital exposure to propidium iodide identifies apoptosis on adherent cells

Cytometry
L ZamaiS Papa

Abstract

Several studies indicate that plasma membrane changes during apoptosis are a general phenomenon. Among the flow cytometric methods to measure apoptosis, the Annexin V assay that detects the membrane exposure of phosphatidylserine (PS) is one of the most commonly used. However, the various treatments used for the detachment of adherent cells generally interfere with the binding of Annexin V to membrane PS, making apoptosis measurement a technical problem. Materials and Methods Apoptosis of different cell lines was investigated by fluorescence microscopy and multiple flow assays designed to assess loss of membrane integrity, translocation of PS, DNA fragmentation, and light scatter changes. Results and Conclusions We show that supravital propidium iodide (PI) assay stains adherent apoptotic cells, allowing flow cytometric quantification. Moreover, supravital exposure to PI without prior permeabilization identifies apoptotic cells as well as Annexin V and permits the simultaneous surface staining by FITC- and PE-conjugated monoclonal antibodies. As in the case of necrotic or permeabilized cells, fluorescence microscopy has revealed that PI staining of apoptotic cells is localized in the nucleus. This suggests that the binding of P...Continue Reading

Citations

Oct 23, 2013·Graefe's Archive for Clinical and Experimental Ophthalmology = Albrecht Von Graefes Archiv Für Klinische Und Experimentelle Ophthalmologie·Jessy SchönfelderChristiane Wetzel
Mar 7, 2008·Apoptosis : an International Journal on Programmed Cell Death·Manuel T SilvaNuno M N dos Santos
Apr 22, 2014·International Journal of Molecular Sciences·Sara SalucciElisabetta Falcieri
May 30, 2014·International Journal of Nanomedicine·Yeliz Utku KoncaRonald N Zuckermann
Apr 1, 2010·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Alessandra RossiAntonio Giordano

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