Fluorescent antibodies and lectins stain intracellular structures in fixed cells treated with nonionic detergent.
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.
D000956:Antigens, Viral; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D004622:Embryo, Mammalian; D005260:Female; D005347:Fibroblasts / Q000032:analysis / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D005557:Formaldehyde; D006801:Humans; D011247:Pregnancy; D011952:Receptors, Concanavalin A / Q000032:analysis*; D011955:Receptors, Drug / Q000032:analysis*; D013539:Simian virus 40
D000956:Antigens, Viral; D011952:Receptors, Concanavalin A; D011955:Receptors, Drug; D005557:Formaldehyde