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How we process trephine biopsy specimens: epoxy resin embedded bone marrow biopsies

Abstract

Improved cytomorphology of semithin resin sections over paraffin wax embedded sections may be important in diagnostic haematopathology. However, resin embedding can make immunohistochemical antigen detection or DNA isolation for clonal gene rearrangement assays difficult. This review describes the processing of bone marrow biopsies using buffered formaldehyde based fixation and epoxy resin embedding, with or without EDTA decalcification. Traditional semithin resin sections are completely rehydrated after etching in home made sodium methoxide solution. Resin elimination allows high resolution staining of tissue components with common histological stains. Efficient antigen retrieval and the Envision-HRP system permit the immunohistological detection of many antigens of diagnostic relevance, with retention of high quality cytomorphology. Furthermore, DNA can be extracted for clonality analysis. The technique can be completed within a similar time period to that of paraffin wax processing with only ∼30% increase in cost. This technique has been used for diagnosis in over 4000 bone marrow biopsies over the past 14 years. By meeting traditional and contemporary demands on the haematopathologist, it offers a powerful alternative to paraffin wax processing for diagnosis and research.

  • DAB, 3,3′-diaminobenzidine tetrahydrochloride
  • EDTA, ethylenediamine tetraacetic acid
  • HRP, horseradish peroxidase
  • PCR, polymerase chain reaction
  • bone marrow trephine biopsy
  • epoxy resin embedding
  • semithin section
  • immunocytochemistry
  • polymerase chain reaction clonality test

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