METHODS FOR IMPROVING THE YIELD AND QUALITY OF METAPHASE PREPARATIONSFOR FISH PROBING OF HUMAN LYMPHOCYTE CHROMOSOMES

Procedures are described for the in vitro culture of human lymphocytes , which have been concentrated by density gradient centrifugation, and for a modified slide-making technique for the Fixed cells. The method yields improved percentages of mitotic cells which are largely synchr onized at harvest. Controlled placement of fixed cells on slides produ ces well-spread metaphase preparations with little background material to interfere with Fluorescence in situ hybridization (FISH) probe pro cedures. The FISH reagents and microscope scanning time required are m inimized by concentrating cells in a defined area of the slide. (C) 19 97 Wiley-Liss, Inc.