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The effect of delay in fixation on HER2 expression in invasive carcinoma of the breast assessed with immunohistochemistry and in situ hybridisation
  1. Andrew H S Lee1,
  2. Heather P Key1,
  3. Jane A Bell2,
  4. Patrick Kumah1,
  5. Zsolt Hodi1,
  6. Ian O Ellis1
  1. 1Department of Histopathology, Nottingham University Hospitals, City Hospital Campus, Nottingham, UK
  2. 2Source Bioscience plc, Nottingham Business Park, Nottingham, UK
  1. Correspondence to Dr A H S Lee, Department of Histopathology, Nottingham University Hospitals, City Hospital Campus, Hucknall Road, Nottingham NG5 1PB, UK; andrew.lee{at}


Aims Accurate assessment of HER2 status is essential for selection of patients for HER2-targeted treatment such as trastuzumab. This study investigated the hypothesis that delayed fixation impairs HER2 assessment.

Methods 9 carcinomas were received fresh, and samples were fixed immediately or put in fixative at time intervals up to 24 h. All carcinomas were scored as 3+ with immunohistochemistry in properly fixed tissue.

Results 2 of 9 carcinomas (95% CIs 6% to 56%) showed reduced immunohistochemical staining with delays in fixation of 1 and 8 h. One carcinoma showed low-level amplification with fluorescence in situ hybridisation (FISH) when properly fixed and was not amplified after delayed fixation. The other carcinomas were amplified at all time points.

Conclusions Delayed fixation impaired HER2 protein expression assessed using immunohistochemistry in 22% of 3+ carcinomas. HER2 amplification assessed using FISH may be less affected.

  • Breast
  • Immunohistochemistry
  • Fixation
  • In Situ Hybridisation

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