Effect of routine paraffin wax processing on cell membrane immunoreactivity in cutaneous tissue.
Author(s): Cerio R, MacDonald DM
Publication: J Clin Lab Immunol, 1986, Vol. 20, Page 97-100
PubMed ID: 2426450 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 influence of fixation duration and discrete stages of tissue processing (dehydration, clearing, paraffin impregnation) on the quality of immunohistochemical staining of the membrane antigen OKT6 in formol saline preserved specimens.
Summary of Findings:
Fixation of divided skin biopsies in formol saline for 15 min to 4 h generated immunostaining comparable in pattern and quality to those observed in OCT embedded frozen sections; fixation for 8 h or longer yielded a reduction in the number of immunopositive cells. Dehydration in ethanol increased background staining, while clearing sections in xylene also substantially reduced immunoreactivity; although these effects were minimized when dehydration and clearing was performed with pre-cooled reagents (4 degrees C). The duration of paraffin impregnation significantly impacted immunoreactivity as well, as immunostaining was reduced after 3 h but not 1 h of paraffin impregnantion at 58 degrees C when employing pronase digestion (10 min at 37 degrees C) of tissue sections.
Biospecimens
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 15 min
30 min
1 h
2 h
4 h
8 h
12 h
24 h
48 h
96 h
Biospecimen Preservation Embedding duration/condition 15 min
30 min
1 h
2 h
3 h
Analyte Extraction and Purification Antigen retrieval Pronase, 10 min at 37 degrees C
Undigested
Biospecimen Preservation Dehydration duration/condition Room temperature
4 degrees C
Biospecimen Preservation Clearing duration/condition Room temperature
4 degrees C