IN-SITU HYBRIDIZATION TO CHROMOSOMES STABILIZED IN GEL MICRODROPS

Conventional chromosome in situ hybridization procedures rely on fixat ion to glass slides followed by microscopic evaluation. This report de scribes the development of a microdrop in situ hybridization (MISH) me thod which facilitates hybridization to chromosomes in suspension. Chr omosomes encapsulated in gel microdrops (GMDs) composed of an agarose matrix withstood stringent hybridization and denaturation conditions. Because of the increased stability, hybridization to encapsulated chro mosomes was detected by how cytometry as well as conventional microsco py. Thus, the MISH method offers a means for chromosome hybridization without slides and may enable identification and isolation of chromoso mes using hybridization rather than nucleic acid binding dyes. (C) 199 5 Wiley-Liss, Inc.