Does rapid microwave fixation improve immunohistochemistry?

Author(s): Azumi N, Joyce J, Battifora H

Publication: Mod Pathol, 1990, Vol. 3, Page 368-72

PubMed ID: 2362942 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of microwave fixation, different fixative solutions, and protease digestion on preservation of tissue specimen morphology and antigens. Skin, endometrium, myometrium, ovary, bladder, colon, ileum, and prostate specimens were examined.

   Summary of Findings:

   In general, morphology of tissues fixed in all four fixatives, through overnight immersion at room temperature or through microwave fixation, was well preserved. Optimal results were obtained with microwave-irradiated Karnovskys and Omnifix-fixed tissues or nonirradiated formaldehyde-fixed tissues. Antigen preservation was generally good for all methods of fixation, however, irradiated and non-irradiated Omnifix and saline-fixed tissues showed the best antigen preservation. Overnight formaldehyde fixation fared the worst with no immunostaining for factor VIII or collagen type IV, and only moderate staining for desmin and keratin. Microwave irradiation of formaldehyde-fixed tissues allowed for excellent immunostaining of all four, and predigestion of non-irradiated formalin-fixed tissue with appropriate proteases led to excellent immunostaining for all but desmin. In contrast, protease digestion of irradiated formalin or Karnovskys-fixed specimens, or of all Omnifix and saline-fixed specimens led to disintegration of tissues.

   Biospecimens

   • Tissue - Skin
   • Tissue - Uterus
   • Tissue - Prostate
   • Tissue - Bladder
   • Tissue - Colorectal
   • Tissue - Small Bowel
   • Tissue - Endometrium
   • Tissue - Ovary

   Preservative Types

   • Other Preservative

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   Morphology	H-and-E microscopy
   Protein	Immunohistochemistry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Method of fixative delivery	Immersion Microwaved
   Immunohistochemistry Specific	Targeted peptide/protein	Keratins Muscle-specific actin Desmin Vimentin Collagen type IV S-100 protein Factor VIII Leukocyte common antigen LN-2 LN-3
   Analyte Extraction and Purification	Protein digestion	Trypsin Pronase
   Biospecimen Preservation	Type of fixation/preservation	Formaldehyde Karnovsky's solution Omnifix Saline

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