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Laser Capture Microdissection: ArcturusXT Infrared Capture and UV Cutting Methods

  • Rosa I. GallagherEmail author
  • Steven R. Blakely
  • Lance A. Liotta
  • Virginia Espina
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 823)

Abstract

Laser capture microdissection (LCM) is a technique that allows the precise procurement of enriched cell populations from a heterogeneous tissue under direct microscopic visualization. LCM can be used to harvest the cells of interest directly or can be used to isolate specific cells by ablating the unwanted cells, resulting in histologically enriched cell populations. The fundamental components of laser microdissection technology are (a) visualization of the cells of interest via microscopy, (b) transfer of laser energy to a thermolabile polymer with either the formation of a polymer–cell composite (capture method) or transfer of laser energy via an ultraviolet laser to photovolatize a region of tissue (cutting method), and (c) removal of cells of interest from the heterogeneous tissue section. Laser energy supplied by LCM instruments can be infrared (810 nm) or ultraviolet (355 nm). Infrared lasers melt thermolabile polymers for cell capture, whereas ultraviolet lasers ablate cells for either removal of unwanted cells or excision of a defined area of cells. LCM technology is applicable to an array of applications including mass spectrometry, DNA genotyping and loss-of-heterozygosity analysis, RNA transcript profiling, cDNA library generation, proteomics discovery, and signal kinase pathway profiling. This chapter describes the unique features of the ArcturusXT laser capture microdissection instrument, which incorporates both infrared capture and ultraviolet cutting technology in one instrument, using a proteomic downstream assay as a model.

Key words

Cancer Laser capture microdissection Tissue Infrared Ultraviolet DNA Molecular profiling Protein RNA Tissue heterogeneity 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Rosa I. Gallagher
    • 1
    Email author
  • Steven R. Blakely
    • 2
  • Lance A. Liotta
    • 1
  • Virginia Espina
    • 1
  1. 1.Center for Applied Proteomics and Molecular MedicineGeorge Mason UniversityManassasUSA
  2. 2.Applied Biosystems/Life Technologies CorporationFoster CityUSA

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