Effect of heat-induced antigen retrieval following inconsistent formalin fixation.

Author(s): Boenisch T

Publication: Appl Immunohistochem Mol Morphol, 2005, Vol. 13, Page 283-6

PubMed ID: 16082257 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 length of formalin fixation and antigen retrieval on IHC staining of 30 tonsil antigens. Freshly frozen tonsil was thawed and placed in 10% neutral buffered formalin for a specified duration.

   Summary of Findings:

   While 26 of the 30 studied antigens were unaffected by length of formalin fixation, CD15, cytokeratin 8, lambda light chains, and p53 showed weaker staining with fixation durations of 4 and 8 d, compared to 12 h, 1 d, or 2 d. Lambda light chains and p53 IHC staining benefited from longer antigen retrieval (60 min) after extended fixation while CD15 and cytokeratin 8 did not. With the exception of CD117, formalin fixation for 3 months yielded weaker IHC staining than in specimens fixed for a shorter duration, despite antigen retrieval at 97 degrees C (20 min). However, antigen retrieval at 121 degrees C (5 min) improved staining for 5 of 7. Background staining was generally absent, although if present, increased slightly with fixation time.

   Biospecimens

   • Tissue - Tonsil

   Preservative Types

   • Formalin

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   Protein	Immunohistochemistry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Time in fixative	12 h 1 d 2 d 4 d 8 d 3 months
   Analyte Extraction and Purification	Antigen retrieval	Target Retrieval Solution 5 min 20 min 60 min
   Analyte Extraction and Purification	Temperature of heat-induced retrieval	97 degrees C 121 degrees C

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