Comparison of formalin-free tissue fixatives: a proteomic study testing their application for routine pathology and research

Arch Pathol Lab Med. 2011 Jun;135(6):744-52. doi: 10.5858/2009-0676-OA.1.

Abstract

Context: Formalin-fixed, paraffin-embedded tissue is the routine processing method for diagnostics practiced in pathology departments worldwide.

Objective: To determine the potential value of non-cross-linking, formalin-free tissue fixation for diagnostics in pathology and proteomic investigations.

Design: We tested 3 commercially available, formalin-free tissue fixatives-FineFIX, RCL2, and HOPE-in lung cancer specimens from 10 patients. The fixatives were evaluated for their effects on tissue morphology, protein recovery, and immunoreactivity for a selected panel of proteins differently expressed in lung cancer, using immunohistochemistry and Western blotting.

Results: Tumor-cell analysis with hematoxylin-eosin worked equally well for all tested fixatives when compared with the standard formalin-fixed, paraffin-embedded procedure. Movat pentachrome stains showed comparable results for the different matrices and cellular proteins analyzed. The RCL2 (P = .01) and HOPE fixatives (P = .03) improved protein recovery when compared with formalin-fixed, paraffin-embedded or frozen tissues. Our data clearly show that the fixatives evaluated influenced immunoreactivity to matched, formalin-fixed, paraffin-embedded lung cancer tissue. In particular, membrane-bound proteins, such as epidermal growth factor receptor EGFR, can be detected more efficiently by immunohistochemistry and Western blotting.

Conclusion: We have demonstrated that formalin-free fixatives have the potential in routine pathology and research to replace formalin in histomorphology and protein preservation.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers, Tumor / analysis
  • Blotting, Western
  • ErbB Receptors / analysis
  • Fixatives*
  • Formaldehyde*
  • Frozen Sections
  • Humans
  • Immunohistochemistry / methods
  • Lung Neoplasms / chemistry*
  • Lung Neoplasms / diagnosis
  • Neoplasm Proteins / analysis*
  • Paraffin Embedding
  • Pathology, Surgical / methods
  • Proteomics / methods
  • Tissue Fixation / methods*

Substances

  • Biomarkers, Tumor
  • Fixatives
  • Neoplasm Proteins
  • Formaldehyde
  • EGFR protein, human
  • ErbB Receptors