6th ESACP Congress, Heidelberg, April 7-11, 1999

A022
LASER CAPTURE MICRODISSECTION - A VERSATILE TOOL FOR THE MOLECULAR ANALYSIS OF TISSUE MICROHETEROGENEITY
Fend F 1,2, Bonner R 3, Emmert-Buck MR 1, Raffeld M 1

(1) Laboratory of Pathology, National Cancer Institute, NIH, Bethesda, MD and (2) Dept. of Pathology, Technical University Munich, Munich, Germany, and (3) Laboratory for Integrated and Medical Biophysics, NICHHD, NIH, Bethesda, MD

Since the first description of laser capture microdissection (LCM) in 1996, technical refinements and the introduction of a commercially available LCM microscope have made this technique an attractive alternative to currently established microdissection methods. Based on the adherence of visually selected cells or tissue areas to an overlying, thermoplastic membrane by focal melting with a low energy infrared laser pulse, LCM allows extremely fast, contact-free isolation of target cells, without requiring manual dexterity or time-consuming micromanipulation. LCM can be applied to a wide range of both fixed and frozen tissues and cytological preparations, employing both routine as well as immunohistochemical stains. High quality DNA, mRNA and proteins can be isolated from appropriately prepared, captured tissue. The detection of genetic alterations including loss of heterozygosity in neoplastic and preneoplastic lesions and the generation of expression libraries and hybridization probes for cDNA microarrays are among the successful applications of LCM. An overview on the methodical aspects of LCM will be given, including practical tips, troubleshooting and new technical developments.