NIH, National Cancer Institute, Division of Cancer Treatment and Diagnosis (DCTD) NIH - National Institutes of Health National Cancer Institute DCTD - Division of Cancer Treatment and Diagnosis

The effects of progressive formaldehyde fixation on the preservation of tissue antigens.

Author(s): Leong AS, Gilham PN

Publication: Pathology, 1989, Vol. 21, Page 266-8

PubMed ID: 2483748 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine the effects of prolonged exposure to formaldehyde and trypsin digestion on immunohistochemistry staining intensity.

Conclusion of Paper

After 3 days of formaldehyde fixation, the staining intensity of some antigens was decreased, and after 7 days, 80% of the antigens tested had reduced staining intensity. Trypsin digestion for 30 min increased the staining intensity of some antigens, but reduced the staining intensity of the lymphocyte antigens. No improvement in staining intensity was found with trypsin digestion longer than 30 min. In conclusion, specimens should be fixed for no more than 6 h, and trypsin digestion for 30 min should be used discriminately.

Studies

  1. Study Purpose

    The purpose of this study was to determine the effect of prolonged exposure to formalin and trypsin on immunohistochemistry staining intensity in lung, kidney, spleen, liver, tonsil, stomach, esophagus, pancreas, thyroid, colon, and prostate. All specimens were fixed in formaldehyde.

    Summary of Findings:

    After 3 days of formaldehyde fixation, the staining intensity of the intermediate filament proteins and neurofilaments were decreased. By 7 days, 80% of the antigens tested had reduced staining intensity. The only antigens that were detectable after 14 days of fixation were S100, prostate specific antigen (PSA), thyroglobulin, and carcinoembryonic antigen (CEA). Trypsin digestion for 30 min increased the staining intensity of cytokeratins, desmin, neurofilament triple proteins, and factor viii related protein, but reduced staining of the lymphocyte antigens. Trypsin digestion beyond 30 min did not further increase the staining and by 75 min some specimens began to disintegrate. In conclusion, specimens should be fixed for no more than 6 h, and trypsin digestion for 30 min should be used discriminately.

    Biospecimens
    Preservative Types
    • Other Preservative
    Diagnoses:
    • Not specified
    Platform:
    AnalyteTechnology Platform
    Protein Immunohistochemistry
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Preservation Time in fixative 6 h
    1 d
    3 d
    7 d
    14 d
    30 d
    Analyte Extraction and Purification Cell/tissue permeabilization 30 min
    45 min
    60 min
    75 min

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