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Immunohistochemistry on old archival paraffin blocks: is there an expiry date?
  1. Federica Grillo1,2,
  2. Martina Bruzzone1,
  3. Simona Pigozzi1,2,
  4. Stefano Prosapio1,
  5. Paola Migliora3,
  6. Roberto Fiocca1,2,
  7. Luca Mastracci1,2
  1. 1Pathology Unit, Department of Surgical Science and Integrated Diagnostics (DISC), University of Genoa, Genoa, Italy
  2. 2IRCCS AOU San Martino IST, Genoa, Italy
  3. 3Pathology Unit, S. Andrea Hospital, Vercelli, Italy
  1. Correspondence to Dr Luca Mastracci, Pathology Unit, Department of Surgical Science and Integrated Diagnostics, University of Genoa, IRCCS AOU San Martino IST, Largo Rosanna Benzi, 10, 16132, Genoa, Italy; mastracc{at}


Few studies have focused on antigen preservation in formalin-fixed and paraffin-embedded (FFPE) tissue in old archival material and additional studies are required, especially considering that these samples are an irreplaceable resource for scientific and clinical research. The purpose of this study is to verify antigen preservation in FFPE tissue samples stored for several decades. From the pathology archives, FFPE blocks were selected dating back to the 1960s, 1970s, 1980s, 1990s, 2000s and 2010. A panel of 12 antibodies was applied and immunoreactivities were compared. While cytoplasmic antigens showed no reduction in immunostaining intensity over time, membrane and nuclear antigens presented reduced staining intensity in older blocks. In particular, the nuclear antigen, Ki67 and CD31 showed the most pronounced antigen decay in the oldest archival blocks. In order to test possible antigen recovery, deep sectioning and lengthening of heat pretreatment were applied. Both strategies partially recover antigenicity, but their simultaneous application shows the best results.

  • KI 67

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  • Handling editor Runjan Chetty

  • Contributors FG and LM planned the research, viewed the slides and wrote the manuscript.

    SPi and SPr performed immunohistochemistry.

    MB and PM identified the cases and provided the FFPE blocks.

    RF supervised and critically reviewed the study and manuscript.

  • Competing interests None declared.

  • Ethics approval Internal departmental ethics committee.

  • Provenance and peer review Not commissioned; externally peer reviewed.

  • Correction notice This paper has been amended since it was published Online First. Owing to a scripting error, some of the publisher names in the references were replaced with ‘BMJ Publishing Group’. This only affected the full text version, not the PDF. We have since corrected these errors and the correct publishers have been inserted into the references.