Introduction of phalloidin labelled with fluorescein isothiocyanate into living polymorphonuclear leukocytes by electroporation

Journal of Biochemical and Biophysical Methods
K HashimotoK Okuda

Abstract

To examine the mechanism by which polymorphonuclear leukocytes (PMNs) move, phalloidin labelled with fluorescein isothiocyanate was introduced into freshly sampled cells by use of an electric-cell fusion system. The best conditions for treatment were three pulses of direct current at 100 V for a pulse duration of 3 microseconds. The treated cells retained their usual motility when observed under a microscope, so the method was suitable for the analysis of motile living cells. We used the method to study PMNs during locomotion, spreading and phagocytosis. In locomotion, fluorescence first appeared at the head of the cell and shifted gradually along the cell margin from head to tail. In spreading, diffuse fluorescence around the marginal part of the cytoplasm was strongest near both the attachment sites and the perinuclear area of the cell and spots of fluorescence appeared in the cytoplasm. In phagocytosis, fluorescence developed from the attachment sites, spread to the entire phagocytizing area of the cytoplasm and disappeared when phagocytosis ended. Cells treated with cytochalasin B were randomly spotted with fluorescence. Freshly sampled cells had diffuse and scattered fluorescence, without the lines observed in fixed cells.

References

Aug 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·T E KreisW Birchmeier
May 1, 1978·The Journal of Cell Biology·S H Zigmond
May 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·J W Sanger
Jun 12, 1987·Biochimica Et Biophysica Acta·H StopperU Zimmermann
Jan 1, 1987·Journal of Cell Science. Supplement·P C Wilkinson
Jun 1, 1986·Cell Structure and Function·H Hama-InabaM Kasai
Jan 1, 1984·Cell and Tissue Research·J J Wolosewick
May 1, 1984·The Anatomical Record·M I RyderR Niederman
Nov 30, 1982·Biochimica Et Biophysica Acta·U Zimmermann
Jan 1, 1982·The Journal of Membrane Biology·U Zimmermann, J Vienken

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Citations

Nov 13, 2008·Cytotechnology·Ti-Fei Yuan
Jan 5, 2000·Journal of Immunological Methods·M Torres, T D Coates

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