Strategies for H-score normalization of preanalytical technical variables with potential utility to immunohistochemical-based biomarker quantitation in therapeutic response diagnostics.
Author(s): Pierceall WE, Wolfe M, Suschak J, Chang H, Chen Y, Sprott KM, Kutok JL, Quan S, Weaver DT, Ward BE
Publication: Anal Cell Pathol (Amst), 2011, Vol. 34, Page 159-68
PubMed ID: 21677381 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 effects of cold ischemia time and fixation duration on IHC staining of breast cancer tissue. 6 tumors were divided into 6 sections each and subjected to different fixation durations, with or without a 24 hour delay to fixation at 4 degrees C in saline. Sections from a breast cancer tissue microarray (TMA) were used as positive controls. H-scores were generated and averaged for 3 regions of interest on whole sections and 2 TMA cores using Aperio's Image Analysis Kit.
Summary of Findings:
For all antigens, H-scores increased with increasing time in fixative, regardless of whether or not specimens were held at 4 degrees C in saline overnight prior to fixation. PARP1 and BRCA1 showed the greatest rates of increase with increasing time in fixative. H-scores between specimens fixed for the same duration with and without a 24 h delay to fixation were not statistically compared, however, lower H-scores were observed for all fixation durations following 24 h cold ischemia time at 4 degrees C compared to H-scores measured with no such delay.
Biospecimens
Preservative Types
- Formalin
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform Protein Immunohistochemistry Protein Tissue microarray Morphology H-and-E microscopy Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Time in fixative 2 h
8 h
48 h
Biospecimen Acquisition Cold ischemia time 0 h
24 h
Immunohistochemistry Specific Targeted peptide/protein BRCA1
SNRPA
SnRNP70
XPF
PARP1
pMK2
ATM