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Laser Capture Microdissection for Protein and NanoString RNA Analysis

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Cell Imaging Techniques

Part of the book series: Methods in Molecular Biology ((MIMB,volume 931))

Abstract

Laser capture microdissection (LCM) allows the precise procurement of enriched cell populations from a heterogeneous tissue, or live cell culture, under direct microscopic visualization. Histologically enriched cell populations can be procured by harvesting cells of interest directly or isolating specific cells by ablating unwanted cells. The basic components of laser microdissection technology are (a) visualization of cells via light 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. The capture and cutting methods (instruments) for laser microdissection differ in the manner by which cells of interest are removed from the heterogeneous sample. Laser energy in the capture method is infrared (810 nm), while in the cutting mode the laser is ultraviolet (355 nm). Infrared lasers melt a thermolabile polymer that adheres to the cells of interest, 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 LCM using an ArcturusXT instrument for downstream protein sample analysis and using an mmi CellCut Plus® instrument for RNA analysis via NanoString technology.

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Acknowledgements

Thanks to the staff of Pathology-Histotechnology Laboratory, SAIC-NCI-Frederick. The work was funded in part by NCI Contract HHSN261200800001E.

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Authors and Affiliations

  1. National Cancer Institute-Frederick/SAIC, Frederick, MD, USA

    Yelena Golubeva & Rosalba Salcedo

  2. Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA, USA

    Claudius Mueller, Lance A. Liotta & Virginia Espina

Authors
  1. Yelena Golubeva
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  2. Rosalba Salcedo
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  3. Claudius Mueller
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  4. Lance A. Liotta
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  5. Virginia Espina
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Corresponding author

Correspondence to Virginia Espina .

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Editors and Affiliations

  1. College of Medicine, Professor, Department of Pathology, University of Vermont, Beaumont Ave 89, Burlington, 05405-1742, Vermont, USA

    Douglas J. Taatjes

  2. , Abt. Zell- und Molekularpathologie, Universität Zürich, Schmelzbergstr. 12, Zürich, 8091, Switzerland

    Jürgen Roth

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© 2012 Springer Science+Business Media, LLC

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Golubeva, Y., Salcedo, R., Mueller, C., Liotta, L.A., Espina, V. (2012). Laser Capture Microdissection for Protein and NanoString RNA Analysis. In: Taatjes, D., Roth, J. (eds) Cell Imaging Techniques. Methods in Molecular Biology, vol 931. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-056-4_12

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  • DOI: https://doi.org/10.1007/978-1-62703-056-4_12

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-055-7

  • Online ISBN: 978-1-62703-056-4

  • eBook Packages: Springer Protocols

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