A low-cost flow cytometric assay for the detection and quantification of apoptosis using an anionic halogenated fluorescein dye.
We describe here a technical improvement of an established colorimetric method used to detect and measure the occurrence of apoptosis in mammalian cells during in vitro cell culture. This assay uses an anionic halogenated fluorescein dye that is taken up by apoptotic cells at the stage of phosphatidylserine externalization. We demonstrate that apoptotic cells stained with this dye can be detected by flow cytometric analysis. Furthermore, we show that the modified method compares well with the standard annexin-V-based apoptosis assay and that it is significantly more cost-effective than the annexin-V assay.
Meyer
Mervin
M
Department of Biotechnology, University of the Western Cape, Bellville, South Africa. mmeyer@mail.biotech.uwc.ac.za
Essack
Magbubah
M
Kanyanda
Stonard
S
Rees
Jasper
J
D017304:Annexin A5 / Q000378:metabolism*; D017209:Apoptosis; D001681:Biological Assay / Q000191:economics; D045744:Cell Line, Tumor; D004396:Coloring Agents / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods; D019793:Fluorescein / Q000737:chemistry / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D019169:Jurkat Cells; D012516:Osteosarcoma / Q000473:pathology; D015203:Reproducibility of Results
D017304:Annexin A5; D004396:Coloring Agents; D005456:Fluorescent Dyes; D019793:Fluorescein