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Optimal processing of bone marrow trephine biopsy: the Hammersmith Protocol

K N Naresh, I Lampert, R Hasserjian, D Lykidis, K Elderfield, D Horncastle, N Smith, W Murray-Brown and G W Stamp

Department of Histopathology, Hammersmith Hospital, London, UK

Correspondence to:
Correspondence to:
K N Naresh
Department of Histopathology, Hammersmith Hospital, Du Cane Road, London W12 0HS, UK; k.naresh{at}imperial.co.uk

Specimens of bone marrow trephine biopsy (BMT) are transported and fixed in acetic acid–zinc–formalin fixative, decalcified in 10% formic acid–5% formaldehyde and processed with other specimens to paraffin-wax embedding. Sections, 1-µm-thick, are cut by experienced histotechnologists and used for haematoxylin and eosin, Giemsa, reticulin silver and other histological stains. Further, all immunohistochemical procedures used in the laboratory, including double immunostaining, can be used on these sections with no or minimal modifications. About 10 000 BMT specimens have been analysed using this procedure since 1997 and diseases involving the bone marrow have been classified successfully. More recently, standardised polymerase chain reaction-based analysis and mRNA in situ hybridisation studies have been conducted. Excellent morphology with good antigen, DNA and RNA preservation is offered by the Hammersmith Protocol.

Abbreviations: AZF, acetic acid–zinc–formalin; BMT, bone marrow biopsy using trephine; EBV, Epstein–Barr virus; EDTA, ethylene diamine tetra acetate; H&E, haematoxylin and eosin; ISH, in situ hybridisation

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Immunocytochemistry and In-Situ Hybridisation on bone marrow trephine biopsy

Lynne Doverty

JCP Online, 14 Feb 2007 [Full text]