Fluorescence in situ hybridization (fish): A user's guide to optimal preparation of cytologic specimens

✍ Scribed by Andrea Abati; Jeffrey S. Sanford; Patricia Fetsch; Francesco M. Marincola; Sandra R. Wolman

• Book ID: 102817586
• Publisher: John Wiley and Sons
• Year: 1995
• Tongue: English
• Weight: 703 KB
• Volume: 13
• Category: Article
• ISSN: 8755-1039
• DOI: 10.1002/dc.2840130518

✦ Synopsis

Fluorescence in situ hybridization (FISH) is a reliable method for tagging centromeric regions of specixc chrornosome.r in interphase nuclei. Not only is FISH useful for chromosome enumeration, but as region-specij7c chromosome probes are developed, the clinical upplicutions and potentials for use by pathologists are extensive, This technique lends itselfparticularly to use in cytology preparations because the cells are disaggregated and monolayer preparations yield excellent technical hybridization results. Over a 7-mo period we processed cytologic samples in an attempt to define and outline a method for optimal specimen processing for FISH use in cell suspensions, techniques applicable to all fresh cytology specimens which can also be used for the processing of surgical pathology aspirates and other material. All samples should be promptly processed to ensure specimen viability, and triaged on an individual basis to ensure preparation of moderately cellular monolayered cytospins. Equivalent nuclear probe signals have been obtained with several sample &ofion methods: air-drying, 95% ethanol, methanol (Dif-Quik fixative), and Carnoy ' s solution. No direrenee was noted in the nuclear probe signals or specimen adhe.sion on positively charged or noncharged slides.

After initial jixation our slides remained at room temperature until FISH was performed, without any adverse effects. A short digestion with proteinase K and subsequent rehybridization yielded positive results on samples that originally yielded poor nuclear probe signals. Diagn Cytopathol 1995;13:486-492. @J 1905 Wilcy-l.iss. he.