Permeabilization of lymphocytes with polyethylene glycol 1000. Discrimination of permeabilized cells by flow cytometry.
The toxicity of polyethylene glycol 1000 (PEG) used similarly as in cell hybridization experiments, has been studied by flow cytometry, measuring the light scattering and fluorescence distributions of PEG-treated human lymphocytes stained with propidium iodide, fluorescein diacetate and acridine orange (AO). The sensitivity of these tests to detect permeabilized, or potentially dead cells, was equal. In addition, PEG proved to interfere with AO staining most likely through the inhibition of its binding to nucleic acids. The decrease of AO fluorescence in cells killed by PEG was unexpected since intercalation of propidium iodide was the same as in alcohol fixed cells. Permeabilization of cells by PEG appears to be an all-or-none phenomenon, accompanied by entrance of PEG into the cells. The findings are described in the context of a review of the currently used flow cytometric techniques to discriminate viable and lethally affected cells; also, the problems of interpretation are discussed.
D000165:Acridine Orange; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002470:Cell Survival; D005434:Flow Cytometry; D006801:Humans; D008027:Light; D008214:Lymphocytes / Q000187:drug effects / Q000502:physiology*; D011092:Polyethylene Glycols / Q000494:pharmacology*; D012542:Scattering, Radiation; D013194:Staining and Labeling
D011092:Polyethylene Glycols; D000165:Acridine Orange