Influence of histochemical stains on quantitative gene expression analysis after laser-assisted microdissection

  • Authors:
    • Ali-Fuat Okuducu
    • Viktor Janzen
    • Jens Claus Hahne
    • Yon Ko
    • Nicolas Wernert
  • View Affiliations

  • Published online on: April 1, 2003     https://doi.org/10.3892/ijmm.11.4.449
  • Pages: 449-453
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Abstract

Laser-assisted microdissection (LAM) allows isolation of specific cell populations for molecular studies. The combination of LAM and of real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) enables generation of quantitative cell-specific gene expression data. Histochemical stains used to identify cells desired for LAM should provide acceptable morphology and not interfere with RNA or with subsequent molecular analysis techniques. To determine a reliable stain for analysing RNA, using the housekeeping gene, RPL13A, we performed quantitative gene expression analysis of laser microdissected cells from prostatic frozen tissues. The frozen sections were histochemically stained with hematoxylin, methyl green, toluidine blue O and May-Grunwald. After laser microdissection real-time quantitative RT-PCR was performed. Methyl green yielded more RT-PCR product than did the other dyes. The lowest yield of amplification was obtained after May-Grunwald staining. Therefore we recommend methyl green for general use in gene expression analysis, especially when handling small amounts of RNA.

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April 2003
Volume 11 Issue 4

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Okuducu A, Janzen V, Hahne JC, Ko Y and Wernert N: Influence of histochemical stains on quantitative gene expression analysis after laser-assisted microdissection. Int J Mol Med 11: 449-453, 2003.
APA
Okuducu, A., Janzen, V., Hahne, J.C., Ko, Y., & Wernert, N. (2003). Influence of histochemical stains on quantitative gene expression analysis after laser-assisted microdissection. International Journal of Molecular Medicine, 11, 449-453. https://doi.org/10.3892/ijmm.11.4.449
MLA
Okuducu, A., Janzen, V., Hahne, J. C., Ko, Y., Wernert, N."Influence of histochemical stains on quantitative gene expression analysis after laser-assisted microdissection". International Journal of Molecular Medicine 11.4 (2003): 449-453.
Chicago
Okuducu, A., Janzen, V., Hahne, J. C., Ko, Y., Wernert, N."Influence of histochemical stains on quantitative gene expression analysis after laser-assisted microdissection". International Journal of Molecular Medicine 11, no. 4 (2003): 449-453. https://doi.org/10.3892/ijmm.11.4.449