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Analysis of Tissue-Specific Gene Expression Using Laser Capture Microdissection

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 585))

Abstract

Epithelial tissues exhibit optimal conditions for studying cellular differentiation since the differentiation status of a single cell can be determined by its distance to the basal membrane. For that reason Laser Capture Microdissection (LCM) may serve as a perfect tool to compare the characteristics of cells that have been collected from different strata of the epithelium. However, as cell boundaries are not visible in untreated tissue sections, samples have to be stained to allow for sufficient structural orientation. This usually results in a considerable reduction of RNA content in the dissected specimen. To circumvent this problem, we have established a modified hematoxylin/eosin staining protocol that concurrently allows visualization of important structures and the subsequent isolation of sufficient RNA amounts to be used for linear amplification and quantitative analyses.

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Acknowledgments

The authors would like to thank Stefan Puschmann for technical support and for providing the figures of capturing epidermal tissue samples.

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© 2010 Humana Press, a part of Springer Science+Business Media, LLC

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Ruetze, M., Lochner, K., Gallinat, S., Knott, A. (2010). Analysis of Tissue-Specific Gene Expression Using Laser Capture Microdissection. In: Turksen, K. (eds) Epidermal Cells. Methods in Molecular Biology, vol 585. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-380-0_14

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  • DOI: https://doi.org/10.1007/978-1-60761-380-0_14

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-379-4

  • Online ISBN: 978-1-60761-380-0

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