IN-SITU HYBRIDIZATION OF CYTOSPIN PREPARATIONS - A RAPID NONISOTOPIC SCREENING METHOD FOR ISOLATED CELLS

A simple and rapid method for light microscopic in situ hybridization on cytospin preparations is described and demonstrated for detection o f viral nucleic acid in a virus-infected cell line. Cells were fixed b y acetone followed by chloroform, denatured by heat, hybridized at 37 C, and hybridized sites detected with a multiple step procedure (prima ry anti-biotin antibody, biotinylated second antibody, streptavidin-pe roxidase). This method can be used for screening studies at the light microscope level, and offers a useful and simple way to determine opti mum hybridization conditions for subsequent electron microscopic inves tigations.