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Minimum Formalin Fixation Time for Consistent Estrogen Receptor Immunohistochemical Staining of Invasive Breast Carcinoma

Neal S. Goldstein MD, Monica Ferkowicz MT(ASCP), PathA(AAPA), Eva Odish HTL(IHQ), Anju Mani MD, Farnaz Hastah MD
DOI: http://dx.doi.org/10.1309/QPHDRB00QXGMUQ9N 86-92 First published online: 1 July 2003


To identify the minimum time necessary for consistent immunohistochemical estrogen receptor (ER) results in our laboratory, we evaluated results in timed fixation blocks and cases with disparate and similar needle core biopsy and partial mastectomy specimens. Tissue sections of 24 ER-positive, invasive breast carcinomas were fixed for 3, 6, 8, and 12 hours and 1, 2, and 7 days. ER values were quantified using the Q score (0–7).

In timed fixation blocks, the mean Q score per block was 2.46 for blocks fixed for 3 hours, 5.75 for blocks fixed for 6 hours, and 6.70 for blocks fixed for 8 hours (P < .001). The difference between the case maximum and mean block Q scores was a plateau of almost 0 at 6 to 8 hours of formalin fixation. For needle core biopsy specimen fixation times, the means for specimens with ER-disparate and ER-similar results were 1.2 and 6.3 hours, respectively (P = .01). The minimum formalin fixation time for reliable immunohistochemical ER results is 6 to 8 hours in our laboratory, regardless of the type or size of specimen.

Key Words:
  • Breast
  • Carcinoma
  • Immunohistochemistry
  • Estrogen
  • Receptor
  • Formalin
  • Fixation