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Fluorescent antibodies and lectins stain intracellular structures in fixed cells treated with nonionic detergent

P Laurila, I Virtanen, J Wartiovaara and S Stenman

Nonionic detergent (NP40) treatment of paraformaldehyde-fixed normal and SV40-transformed human fibroblasts resulted in intracellular penetration of two chosen fluorescent antibodies and Concanavalin A (Con A). After the detergent treatment nuclear SV40 T antigen, cytoplasmic fibronectin glycoprotein and Con A binding sites could be visualized in fluorescence microscopy. The lowest NP40 concentration which made fixed cells permeable was 0.05%. The morphology of cells was preserved better by this new method than by conventional fixation methods, such as acetone treatment. In scanning electron microscopy the surface of the fixed NP40-treated cells had only small rugosities and fine pores. The subsurface cytoskeleton especially was well preserved and had a more distinct fine structure. The improved morphology made it possible to detect a similar distribution of fibronectin and Con A binding sites in the perinuclear endoplasmic reticulum regions.

Volume 26, Issue 4, pp. 251-257, 04/01/1978
Copyright © 1978 by The Histochemical Society


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P Laurila and I Leivo
Basement membrane and interstitial matrix components form separate matrices in heterokaryons of PYS-2 cells and fibroblasts
J. Cell Sci., January 1, 1993; 104(1): 59 - 68.
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