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Detection of drug-induced apoptosis by flow cytometry after alkaline extraction of ethanol fixed cells

1996; Springer Science+Business Media; Volume: 2; Issue: 1-2 Linguagem: Inglês

10.1007/bf02893956

1532-2807

Rudolf Mihalik, Ferenc Uher, Éva Pocsik, Lajos Berczi, M. Benczúr, László Kopper,

Bacteriophages and microbial interactions

A new flow cytometric method was developed to detect apoptotic cells with fragmented DNA and to determine cell cycle distribution of viable cells, in the same sample, by propidium iodide staining. Apoptosis, in HT58 human B lymphoma cells, was induced by etoposide and/or by staurosporine. Using appropriate alkaline solutions (between 1-10 mN NaOH in 150 mM saline) followed by neutralization with buffer solution, the fragmented DNA can be extracted quantitatively from ethanol fixed cells. Further, good resolution of the cell cycle distribution can be obtained in unimpaired cells without RNase treatment. Furthermore, unlike the widely used hypotonic-detergent extraction of unfixed cells, the suggested extraction method can prevent drug-induced disintegration of dead cells when karyorrhexis occurs.