Solid Tissue Flow Cytometry

Sajeed Iqbal, Haematology

Sajeed Iqbal

Flow cytometry immunophenotyping has been reported as a useful adjunct to conventional histological and immunohistochemical findings in the diagnosis and classification of lymphomas. Immunophenotyping has become a field which has had a vast impact in the diagnosis of lymphoma. There is an array of monoclonal antibodies which are available and that can differentiate between various epitopes based on the cell surface or cytoplasm. Immunophenotyping has been applied to specimens such as fine needle aspiration biopsies, effusions, and cerebrospinal fluids. This has been key to distinguishing monoclonal involvement and reactive infiltration.
The aim of this study was to assess the feasibility of using flow cytometry in the analysis of solid tissue, such as lymph nodes and tonsils. If successful this would aid in the diagnosis of haematological malignancies, such as lymphomas.

Fresh tonsils and lymph nodes from consenting patients were collected from theatre. Several methods of  mechanical and chemical cell dissociation were tested to optimise cell suspension preparation. The single cell suspension will then be analysed by 3-4 colour flow cytometry. The whole blood lysis procedure will be used to analyse the specimen. Standard lymphoma panel CD markers will be used in the analysis of the specimen.

The results of this study show that cell dissociation by using either mechanical methods (MediMachine) or the Perfusion Method, followed by flow cytometric analysis is a good method for the assessment of lymphoid cells. This method could be introduced as part of the routine work up of solid tissue sample suspicious for lymphoma