Summary
Gene expression analysis requires a sound basis of cell material to obtain realistic results. Tissue, however, consists of diverse types of cells, which often differentially express target genes, so that cell populations need to be selected. If tissue diversity is moderate and negligible, manual microdissection can be the cost-efficient method of choice. In contrast, the advantage of laser microdissection is a very exact selection down to the level of a single cell, but often with a considerable time needed to get enough material for the following analyses. The latter issue and the method of tissue preparation needed for laser microdissection are the main problems to solve if RNA, highly sensitive to degradation, shall be analyzed. This method focuses on optimized laser microdissection procedures for RNA analysis, drawing on the very heterogeneous tissue of prostatic adenocarcinoma.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Micke, P., Ostman, A., Lundeberg, J., and Ponten, F. (2005) Laser-assisted cell microdissection using the PALM system. Methods Mol.Biol. 293, 151–166.
Schutze, K., Niyaz, Y., Stich, M., and Buchstaller, A. (2007) Noncontact laser microdissection and catapulting for pure sample capture. Methods Cell. Biol. 82, 649–673.
Espina, V., Milia, J., Wu, G., Cowherd, S., and Liotta, L.A. (2006) Laser capture microdissection. Methods Mol. Biol. 319, 213–229.
Espina, V., Wulfkuhle, J.D., Calvert, V.S., VanMeter, A., Zhou, W., Coukos, G., et al. (2006) Laser-capture microdissection. Nat. Protoc. 1, 586–603.
Espina, V., Heiby, M., Pierobon, M., and Liotta, L.A. (2007) Laser capture microdissection technology. Expert Rev. Mol. Diagn. 7, 647–657.
Anslinger, K., Mack, B., Bayer, B., Rolf, B., and Eisenmenger, W. (2005) Digoxigenin labelling and laser capture microdissection of male cells. Int. J. Legal Med. 119, 374–377.
Kolble, K. (2000) The LEICA microdissection system: design and applications. J. Mol. Med. 78, B24–B25.
Vega, C.J. (2008) Laser microdissection sample preparation for RNA analyses. Methods Mol. Biol. 414, 241–252.
Murray, G.I. (2007) An overview of laser microdissection technologies. Acta Histochem. 109, 171–176.
Walker, J.M. (ed.) (2003, 2005–2008) Methods in Molecular Biology. Humana, Totowa, NJ.
Ornstein, D.K., Gillespie, J.W., Paweletz, C.P., Duray, P.H., Herring, J., Vocke, C.D., et al. (2000) Proteomic analysis of laser capture microdissected human prostate cancer and in vitro prostate cell lines. Electrophoresis 21, 2235–2242.
Richardson, A.M., Woodson, K., Wang, Y., Rodriguez-Canales, J., Erickson, H.S., Tangrea, M.A., et al. (2007) Global expression analysis of prostate cancer-associated stroma and epithelia. Diagn. Mol. Pathol. 16, 189–197.
Acknowledgments
The author is very grateful to Cornelia Stelzer and Sabine Becker for technical assistance and photography.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Rabien, A. (2009). Laser Microdissection. In: Grützmann, R., Pilarsky, C. (eds) Cancer Gene Profiling. Methods in Molecular Biology, vol 576. Humana Press. https://doi.org/10.1007/978-1-59745-545-9_3
Download citation
DOI: https://doi.org/10.1007/978-1-59745-545-9_3
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-934115-76-3
Online ISBN: 978-1-59745-545-9
eBook Packages: Springer Protocols