Laser-assisted microdissection of membrane-mounted paraffin sections for polymerase chain reaction analysis: Identification of cell populations using immunohistochemistry and in situ hybridization

Lise Mette Gjerdrum, Ilze Lielpetere, Lars Melholt Rasmussen, Knud Bendix, Stephen Hamilton-Dutoit

Research output: Contribution to journalArticlepeer-review

33 Citations (Scopus)

Abstract

Laser microbeam microdissection (LMM) is an increasingly important method for obtaining pure cell samples for genetic and proteomic analysis. Immuno-histochemistry (IHC) and in situ hybridization (ISH) are useful techniques for targeting specific cell populations for microdissection but are difficult to apply with the tissue support membranes often used during LMM. Using detection of cytokeratins and Epstein-Barr virus gene products in head and neck carcinoma as a model, we describe optimized protocols for membrane and section preparation and for low temperature antigen retrieval that allow IHC and ISH to be used reliably on membrane mounted paraffin tissue sections. Visualization of cellular targets was markedly improved by staining and this could be further improved using a variety of optical media before microdissection. Tissue fragments thus stained were suitable for subsequent polymerase chain reaction analysis of extracted DNA using standard techniques. These IHC and ISH procedures are generally applicable and will be useful for detecting a wide range of antigens and nucleic acids in paraffin sections in conjunction with LMM.

Original languageEnglish
Article number60659
Pages (from-to)105-110
Number of pages6
JournalJournal of Molecular Diagnostics
Volume3
Issue number3
DOIs
Publication statusPublished - 2001
Externally publishedYes

Field of Science

  • 3.1 Basic medicine
  • 3.2 Clinical medicine

Publication Type

  • 1.1. Scientific article indexed in Web of Science and/or Scopus database

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