Delay to formalin fixation effect on breast biomarkers.
Author(s): Khoury T, Sait S, Hwang H, Chandrasekhar R, Wilding G, Tan D, Kulkarni S
Publication: Mod Pathol, 2009, Vol. 22, Page 1457-67
PubMed ID: 19734848 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to investigate the effect of cold ischemia duration (0-24 h) prior to fixation on FISH staining for HER-2 and on ER, PR and HER-2 protein levels as determined by immunohistochemistry. Breast carcinomas were procured via lumpectomy from 10 patients, processed immediately and fixed as soon as possible (within 2-5 min) or placed in 0.9% NaCl and stored for varying durations (10 min-8 h) at room temperature or at 4°C overnight. Specimens were fixed in 10% neutral buffered formalin for 12-32 h and were uniformly processed. FFPE specimens were then assembled into tissue microarrays using 0.6 mm cores.stored post surgical excision in 0.9% NaCl for the specified cold ischemia time.
Summary of Findings:
Cold ischemia time of more than 1 hour resulted in weakened or non-uniform HER-2 fluorescent in situ hybridization (FISH) staining or poor nuclear /cellular resolution such that at 8 h only 1 sample was uncompromised. This effect was not observed for the chromosome 17 centromere signal potentially due to variability of DNA to autolysis. Cold ischemia prior to immunohistochemistry of estrogen (ER) and progesterone receptors (PR) resulted in a non significant trend toward decreased staining intensity and decreased percentage of immunopositive cells. The effect was first noted at 2 h for ER and 1 h for PR, and in both cases resulted in a declining Q score that altered prognosis from good to intermediate for several cases. HER-2 immunostaining was variable in most cases and consequently unscored. Storage overnight at 4 degrees C resulted in similar finding to storage for 8 h at room temperature. Based on these finding the authors suggest all specimens should be fixed within 1 h of surgical excision for proper hormone receptor and HER-2 status determination.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform DNA FISH DNA Tissue microarray Protein Immunohistochemistry Protein Tissue microarray Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Acquisition Cold ischemia time 0
10 min
30 min
1 h
2 h
4 h
8 h
24 h
Storage Storage temperature Room temperature
4 degrees C
Immunohistochemistry Specific Targeted peptide/protein Estrogen receptor
Progesterone receptor
HER-2