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Immunophenotyping (Flow Cytometry)

 

Scientist in Charge

  • David Bloxham

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About

Flow cytometry is performed primarily using 8 colour panels (progression to 12 colour panels is underway) in addition to the forward and side scatter properties of the cells. Antibody panels are designed to identify the main cell populations present in the sample and to focus on particular populations of interest with respect to lineage assignment and stage of differentiation. Aberrant antigen expression or asynchrony can be a marker of abnormality. Additionally, detection of light chain restriction within B-cell populations can be a very powerful tool for the detection of clonal B-lymphoid populations.

Bone marrow and tissue samples are usually reported with the percentage of a population of interest together with their phenotype. When reporting peripheral blood lymphoid populations this usually includes absolute values.

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Specimens required

  • Blood (10ml EDTA) and/or bone marrow (1–2 mL EDTA) – this can be the sample provided for morphology:

  • If sending peripheral blood or bone marrow, please enclose an unstained/stained smear preparation for evaluation

  • If bone marrow aspirates are unobtainable or inadequate, bone marrow trephines (placed in sterile saline or RPMI) can be disaggregated for flow cytometric analysis*

  • Fresh tissue biopsy* or fine needle aspirate (FNA) in sterile saline or RPMI

  • Body fluids (in sterile tube) e.g. CSF, pleural fluid, lavage

  • Apheresis collection and/or cord blood in EDTA or ACD

*This must always be performed as a secondary investigation. The first bone marrow trephine biopsy or tissue biopsy must always be placed in formalin for primary haematopathology reporting.

Sample storage

  • Analyse within 48 hours of collection

  • Keep at 4C before despatch


Reporting

  • The immunophenotyping report will include the clinical details from the request form and will state the sample type and cell population analysed.

  • The percentage of the cells with the phenotype of interest will be reported with respect to the whole population.

  • An overall interpretation and conclusion will be provided. Phenotypes, which may be useful for monitoring of minimal residual disease or response evaluation, will be indicated.

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