Electropermeabilization and fluorescent tracer exchange: the role of whole-cell capacitance

Cytometry
V L SukhorukovU Zimmermann

Abstract

Transmembrane crossing of charged fluorescent tracers such as propidium iodide (PI) and carboxyfluorescein+ (CF) can be used to quantitate membrane permeabilization. Murine myeloma Sp2/0-Ag14 cells were loaded with CF (0.1 fmol/cell) before electropulsation (0.5-3.0 kV/cm, 40 microseconds) in medium containing 25-50 micrograms/ml PI at 21-23 degrees C. Cytograms of PI vs. CF fluorescence showed three readily distinguishable subpopulations: 1) intact living cells with CF but without PI (these form > 95% of the prepulsed population), 2) transiently electropermeabilized but resealed cells showing both CF and low-level PI fluorescence, and 3) permanently permeabilized cells without CF but with very high PI fluorescence. Despite the ready influx of PI, the efflux of CF from transiently permeabilized cells was negligible and was insensitive to pulse parameters; however, electrically killed cells (subpopulation 3) lost all CF fluorescence and probably lost their cytoplasm. This difference in transmembrane passage of the dyes is best explained by binding of intracellular CF to macromolecules (and/or organelles). In isotonic "pulse medium," the membranes resealed after electropulsing with a time constant (tau R) of about 2 min. In 150 m...Continue Reading

References

Jul 16, 1979·The Journal of Membrane Biology·R BenzU Zimmermann
Sep 1, 1975·The Journal of Cell Biology·S KnuttonC A Pasternak
Mar 1, 1989·International Journal of Cardiology·A M Scammell, R Arnold
Jun 13, 1986·Biochimica Et Biophysica Acta·Q F Ahkong, J A Lucy
Jun 27, 1985·Biochimica Et Biophysica Acta·K Schwister, B Deuticke
Jan 1, 1966·Proceedings of the National Academy of Sciences of the United States of America·B Rotman, B W Papermaster
Feb 1, 1980·Quarterly Reviews of Biophysics·J Seelig, A Seelig
Jan 1, 1981·Cytometry·K K Meisingset, H B Steen
Sep 1, 1980·Cytometry·E HamoriT M Jovin
Oct 1, 1994·The Journal of Membrane Biology·V L SukhorukovU Zimmermann
Feb 1, 1993·The Journal of Membrane Biology·V L SukhorukovU Zimmermann
Jun 8, 1993·Biochimica Et Biophysica Acta·S Orlowski, L M Mir

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Citations

Dec 18, 2001·Cytometry·H MussauerU Zimmermann
Aug 14, 2003·Experimental Cell Research·Kilian J MüllerVladimir L Sukhorukov
Apr 18, 2008·Journal of Biological Engineering·Liju YangRashid Bashir
Aug 14, 2001·Biochemical and Biophysical Research Communications·C S Djuzenova, M Flentje
Jul 11, 2014·Expert Opinion on Drug Delivery·Maša Kandušer, Marko Ušaj
Oct 13, 2009·Journal of Controlled Release : Official Journal of the Controlled Release Society·Z FanC X Deng
Nov 30, 2013·Scientific Reports·Lea RemsGorazd Pucihar
Jun 24, 2004·Experimental Cell Research·Yulia AntovRafi Korenstein
Sep 8, 2012·Biochimica Et Biophysica Acta·Jianbo LiHao Lin

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