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6th ESACP Congress, Heidelberg, April 7-11, 1999 |
A022
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.
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