Optimal processing of bone marrow trephine biopsy: the Hammersmith Protocol.

Area of Science:

• Histopathology
• Molecular Biology
• Biomedical Science

Background:

• Bone marrow trephine biopsy (BMT) analysis is crucial for diagnosing hematological malignancies.
• Standard fixation and processing methods can compromise tissue morphology and molecular preservation.

Purpose of the Study:

• To present the Hammersmith Protocol for processing BMT specimens.
• To evaluate the protocol's effectiveness in preserving morphology and molecular analytes for various downstream analyses.

Main Methods:

• BMT specimens are fixed in acetic acid-zinc-formalin, decalcified in formic acid-formaldehyde, and embedded in paraffin wax.
• 1-micrometer sections are stained using H&E, Giemsa, reticulin silver, and immunohistochemistry.
• The protocol supports polymerase chain reaction (PCR)-based analysis and mRNA in situ hybridization.

Main Results:

• The Hammersmith Protocol has been used to analyze approximately 10,000 BMT specimens since 1997.
• Successful classification of bone marrow diseases has been achieved using this method.
• Excellent morphology with good antigen, DNA, and RNA preservation is consistently observed.

Conclusions:

• The Hammersmith Protocol provides superior preservation of bone marrow biopsy specimens.
• It enables comprehensive histopathological and molecular analyses for accurate disease diagnosis and classification.