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
Conclusion of Paper
Studies
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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
- Tissue - Lung
- Tissue - Spleen
- Tissue - Liver
- Tissue - Tonsil
- Tissue - Prostate
- Tissue - Esophagus
- Tissue - Pancreas
- Tissue - Thyroid Gland
- Tissue - Colorectal
- Tissue - Kidney
- Tissue - Stomach
Preservative Types
- Other Preservative
Diagnoses:
- Not specified
Platform:
Analyte Technology Platform Protein Immunohistochemistry Pre-analytical Factors:
Classification Pre-analytical Factor Value(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