Cancer Diagnosis Program | Biorepositories and Biospecimen Research Branch

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Brief formalin fixation and rapid tissue processing do not affect the sensitivity of ER immunohistochemistry of breast core biopsies.

Author(s): Sujoy V, Nadji M, Morales AR

Publication: Am J Clin Pathol, 2014, Vol. 141, Page 522-6

PubMed ID: 24619753 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   This study compared the IHC staining intensity and frequency of breast specimens fixed for 30 min-1 week and processed using a microwave-assisted processor with specimens fixed using the American Society of Clinical Oncology/College of American Pathologists (ASCO/CAP) guidelines of 12-48 h and processed using a conventional processor. Five 1.0 x 0.2 x 0.2 cm core specimens were obtained from each of 22 mastectomy specimens from patients with infiltrating ductal carcinoma. Specimens were fixed in 10% NBF for 30 min, 60 min, 24 h, or 1 week and processed in a microwave-assisted processor or fixed following the ASCO/CAP guidelines for 12-48 h and processed in a conventional tissue processor. Paraffin-embedded blocks were stored at room temperature until all specimens could be sectioned. For each case, slides were created with 3µm sections from specimens fixed for 30 min, 60 min and 24 h and slides with sections of specimens fixed for 1 week or following the ASCO/CAP guidelines (12-48 h). A Q score was assigned by two different pathologists for IHC staining by adding an intensity score (0-3) and a proportion of positive nuclei score (0-4) resulting in scores of 0-7 with 0 being negative, 1-5 being moderately positive, and 6-7 strongly positive.

   Summary of Findings:

   The percentage of ER positive nuclei and the intensity of ER IHC staining were comparable among all 17 ER positive specimens regardless of fixation duration (30 min-1 week) or use of microwave-assisted processor rather than a conventional processor. Further, with the exception of 1 core fixed for 24 h, the Q scores assigned based on the proportion of cells stained and staining intensity were “7” for all 5 cores from each of the 17 ER positive cases, with the remaining core scoring a “6”. The 5 cases that were found to be ER negative were ER negative in all cores, regardless of fixation time. The Q scores determined by the two different observers matched in all cases.

   Biospecimens

   • Tissue - Breast

   Preservative Types

   • Formalin

   Diagnoses:

   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   Morphology	H-and-E microscopy
   Protein	Immunohistochemistry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Time in fixative	30 min 60 min 24 h 1 week 12-48 h
   Immunohistochemistry Specific	Targeted peptide/protein	ER
   Biospecimen Preservation	Duration of tissue/ specimen processing	Rapid microwave-assisted processor Overnight conventional processor

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