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Paraffin Section Immunotyping of Leukaemias (pp. 95-111) $25.00
Authors:  Juneja, S. (Royal Melbourne Hospital, Melbourne, Australia) Westerman, D. and Seymour, J. F. (Peter MacCallum Cancer Center, Melbourne, Australia)
Abstract:
To better understand underlying biological heterogeneity and optimize management, leukaemias are increasingly being characterized by multiple methodologies. In addition to morphology, cytogenetics and molecular studies, immunophenotyping is important for both the accurate diagnosis and the detection of residual disease. Immunophenotyping can be performed by flow cytometry (FCM) in most cases and is the preferred method whenever a fresh tissue samples is available; however, there are circumstances where the leukaemias need to be characterised immunophenotypically in paraffin tissue sections (paraffin section immunophenotyping PSI). The circumstances in which PSI is useful include, dry or blood tap of the bone marrow, difficulty in establishing lineage by FCM and detection of residual disease post-chemotherapy not identified by FCM; it is also applicable where leukaemia was not suspected in the differential diagnosis and appropriate samples for FCM were not taken. PSI also has the advantage with good preservation of cellular morphology, which can be visualised, in tissue sections unlike FCM. PSI is applicable in tissues fixed in formalin, Bouin's or B5 fixatives. The most commonly used method is the labelled streptavidin-biotin peroxidase method with diaminobenzidine or aminoethyl carbozole used as a chromogen. The panel of antibodies utilized varies with the subtype of leukaemia under investigation. In the context of an acute leukaemia it generally includes non-lineage specific markers CD34 and TdT and lineage specific markers including myeloperoxidase (granulocytic), CD3 (T lymphoid) and CD79a (B lymphoid). In chronic leukaemias depending upon whether the cell of origin is B, T or NK lineage appropriate combination of antibodies can be used. In conclusion PSI has a definite role in the diagnosis, characterization and monitoring of residual disease in selected cases of leukaemia. Its role continues to expand as the available antibody panel grows the technique is automated and becomes more widely available. 


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Paraffin Section Immunotyping of Leukaemias (pp. 95-111)