title: Laser capture microdissection: should an ultraviolet or infrared laser be used?
abstract: Laser capture microdissection (LCM) is a well-established cell separation technique. It combines microscopy with laser beam technology and allows targeting of specific cells or tissue regions that need to be separated from others. Consequently, this biological material can be used for genome or transcriptome analyses. Appropriate methods of sample preparation, however, are crucial for the success of downstream molecular analysis. The aim of this study was to objectively compare the two main LCM systems, one based on an ultraviolet (UV) laser and the other based on an infrared (IR) laser, on different criteria ranging from user-friendliness to sample quality. The comparison was performed on two types of samples: peripheral blood mononuclear cells and blastocysts. The UV laser LCM system had several advantages over the IR laser LCM system. Not only does the UV system allow faster and more precise sample collection, but also the obtained samples-even single cell samples-can be used for DNA extraction and downstream polymerase chain reaction (PCR) applications. RNA-based applications are more challenging for both LCM systems. Although sufficient RNA can be extracted from as few as 10 cells for reverse transcription quantitative PCR (RT-qPCR) analysis, the low RNA quality should be taken into account when designing the RT-qPCR assays.
authors: Laboratory for Pharmaceutical Biotechnology, Faculty of Pharmaceutical Sciences, Ghent University, B-9000 Ghent, Belgium.
authors: Mado
authors: M
authors: Vandewoestyne
authors: Karen
authors: K
authors: Goossens
authors: Christian
authors: C
authors: Burvenich
authors: Ann
authors: A
authors: Van Soom
authors: Luc
authors: L
authors: Peelman
authors: Dieter
authors: D
authors: Deforce
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology; D002417:Cattle; D004247:DNA; D007259:Infrared Rays; D061346:Laser Capture Microdissection / Q000295:instrumentation* / Q000379:methods; D007834:Lasers / Q000145:classification*; D007963:Leukocytes, Mononuclear / Q000166:cytology; D016133:Polymerase Chain Reaction / Q000379:methods; D012313:RNA; D014466:Ultraviolet Rays
chemical_list: D012313:RNA; D004247:DNA
keywords: DNA; IR; Laser capture microdissection; RNA; UV