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Laser-Assisted Microdissection of Membrane-Mounted Tissue Sections

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Laser Capture Microdissection

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 293))

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Abstract

Biological tissues (in particular those affected by disease) are inherently complex mixtures of different cell types and matrices. This heterogeneity can complicate the interpretation of molecular biological studies performed on whole-tissue extracts if the precise cellular origin of the molecules being tested is not known. Laser-assisted micro dissection. (LAM) has emerged as a leading histological technique for obtaining samples enriched for specific target cell populations or tissue components for subsequent molecular (especially polymerase chain reaction-based) analysis. This method allows the identification and study of target-specific molecular alterations in heterogeneous specimens, and enables more accurate detection and quantification of target molecules. In this chapter, we focus on tissue microdissection performed with an ultraviolet laser system and describe protocols for the basic procedure and for handling of the samples.

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References

  1. Bohm, M., Wieland, I., Schutze, K., and Rubben, H. (1997) Microbeam MOMeNT: non-contact laser microdissection of membrane-mounted native tissue. Am. J. Pathol. 151, 63–67.

    PubMed  CAS  Google Scholar 

  2. Aubele, M., Zitzelsberger, H., Schenck, U., Walch, A., Hofler, H., and Werner, M. (1998) Distinct cytogenetic alterations in squamous intraepithelial lesions of the cervix revealed by laser-assisted microdissection and comparative genomic hybridization. Cancer 84, 375–379.

    Article  PubMed  CAS  Google Scholar 

  3. Luo, L., Salunga, R. C., Guo, H., et al. (1999) Gene expression profiles of laser-captured adjacent neuronal subtypes. Nat. Med. 5,117–122.

    Article  PubMed  CAS  Google Scholar 

  4. Weber, R. G., Scheer, M., Born, I. A., et al. (1998) Recurrent chromosomal imbalances detected in biopsy material from oral premalignant and malignant lesions by combined tissue microdissection, universal DNA amplification, and comparative genomic hybridization. Am. J. Pathol. 153, 295–303.

    Article  PubMed  CAS  Google Scholar 

  5. Bohm, M. and Wieland, I. (1997) Analysis of tumour-specific alterations in native specimens by PCR: How to procure the tumour cells! (review). Int. J. Oncol. 10, 131–139.

    Google Scholar 

  6. Emmert-Buck, M. R., Bonner, R. F., Smith, P. D., et al. (1996) Laser capture microdissection. Science 274, 998–1001.

    Article  PubMed  CAS  Google Scholar 

  7. Schutze, K. and Lahr, G. (1998) Identification of expressed genes by laser-mediated manipulation of single cells. Nat. Biotechnol. 16, 737–742.

    Article  PubMed  CAS  Google Scholar 

  8. Lahr, G. (2000) RT-PCR from archival single cells is a suitable method to analyze specific gene expression. Lab. Invest. 80, 1477–1479.

    PubMed  CAS  Google Scholar 

  9. Banks, R. E., Dunn, M. J., Forbes, M. A., et al. (1999) The potential use of laser capture microdissection to selectively obtain distinct populations of cells for proteomic analysis-preliminary findings. Electrophoresis 20, 689–700.

    Article  PubMed  CAS  Google Scholar 

  10. Gjerdrum, L. M., Lielpetere, I., Rasmussen, L. M., Bendix, K., and Hamilton-Dutoit, S. (2001) Laser-assisted microdissection of membrane-mounted paraffin sections for polymerase chain reaction analysis: identification of cell populations using immunohistochemistry and in situ hybridization. J.Mol. Diagn. 3,105–110.

    Article  PubMed  CAS  Google Scholar 

  11. Specht, K., Richter, T., Muller, U., Walch, A., Werner, M., and Hofler, H. (2001) Quantitative gene expression analysis in microdissected archival formalin-fixed and paraffin-embedded tumor tissue. Am. J. Pathol. 158, 419–429.

    Article  PubMed  CAS  Google Scholar 

  12. Burton, M. P., Schneider, B. G., Brown, R., Escamilla-Ponce, N., and Gulley, M. L. (1998) Comparison of histologic stains for use in PCR analysis of microdissected, paraffin-embedded tissues. Biotechniques 24, 86–92.

    PubMed  CAS  Google Scholar 

  13. Murase, T., Inagaki, H., and Eimoto, T. (2000) Influence of histochemical and immunohistochemical stains on polymerase chain reaction. Mod. Pathol. 13, 147–151.

    Article  PubMed  CAS  Google Scholar 

  14. Serth, J., Kuczyk, M. A., Paeslack, U., Lichtinghagen, R., and Jonas, U. (2000) Quantitation of DNA extracted after micropreparation of cells from frozen and formalin-fixed tissue sections. Am. J. Pathol. 156,1189–1196.

    Article  PubMed  CAS  Google Scholar 

  15. Ehrig, T., Abdulkadir, S. A., Dintzis, S. M., Milbrandt, J., and Watson, M. A. (2001) Quantitative amplification of genomic DNA from histological tissue sections after staining with nuclear dyes and laser capture microdissection. J. Mol. Diagn. 3, 22–25.

    Article  PubMed  CAS  Google Scholar 

  16. Kuppers, R., Zhao, M., Hansmann, M. L., and Rajewsky, K. (1993) Tracing B cell development in human germinal centres by molecular analysis of single cells picked from histological sections. EMBO J. 12,4955–4967.

    PubMed  CAS  Google Scholar 

  17. Lehmann, U. and Kreipe, H. (2001) Real-time PCR analysis of DNA and RNA extracted from formalin-fixed and paraffin-embedded biopsies. Methods 25, 409–418.

    Article  PubMed  CAS  Google Scholar 

  18. Dietmaier, W., Hartmann, A., Wallinger, S., et al. (1999) Multiple mutation analyses in single tumor cells with improved whole genome amplification. Am. J. Pathol. 154, 83–95.

    Article  PubMed  CAS  Google Scholar 

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© 2005 Humana Press Inc.

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Mette, L., Hamilton-Dutoit, S. (2005). Laser-Assisted Microdissection of Membrane-Mounted Tissue Sections. In: Murray, G.I., Curran, S. (eds) Laser Capture Microdissection. Methods in Molecular Biology™, vol 293. Humana Press. https://doi.org/10.1385/1-59259-853-6:127

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  • DOI: https://doi.org/10.1385/1-59259-853-6:127

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-260-5

  • Online ISBN: 978-1-59259-853-3

  • eBook Packages: Springer Protocols

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