Effect of formalin tissue fixation and processing on immunohistochemistry

Am J Surg Pathol. 2000 Jul;24(7):1016-9. doi: 10.1097/00000478-200007000-00014.


Although immunohistochemistry is routinely performed by many pathology laboratories, its standardization still lags behind. A major cause of variation in the reproducibility of immunohistochemical staining is induced by tissue fixation and, to a lesser degree, tissue processing. This report, stemming from the first meeting of the International Consensus Group on Standardization and Quality Control (ICGSQC) in Nice, France, summarizes the problem and suggests solutions to begin to achieve standardization of fixation and processing. Most laboratories use neutral-buffered formalin (10%) for tissue fixation which introduces cross-links, whereas coagulative fixatives are less popular. Problems with formalin fixation comprise delay of fixation and variations in the duration of the fixation mainly. Solutions to these problems could be to start fixation soon (<30 min) after surgical removal of the tissue and to avoid overfixation (>24-48 hrs). For tissue processing, the most important problem is inadequate tissue dehydration prior to paraffin embedding. This can be prevented by preparing all solutions freshly every week, depending on the volume of tissue processed. If consistently applied, these procedures could eliminate some of the sources of variation in immunohistochemical stains.

Publication types

  • Review

MeSH terms

  • Artifacts
  • Consensus Development Conferences as Topic
  • Cross-Linking Reagents
  • Fixatives*
  • Formaldehyde*
  • Humans
  • Immunohistochemistry / methods*
  • Paraffin Embedding
  • Tissue Fixation / standards*


  • Cross-Linking Reagents
  • Fixatives
  • Formaldehyde