Immunophenotyping (flow cytometry)

Scientist in Charge:        Dr David Bloxham

Telephone Number:        (01223) 217132

 

Immunophenotyping is mainly based on 8-colour flow cytometry, which also includes the use of the physical properties (forward scatter and side scatter) of the cells, together with their antigenic make-up. Antibody panels are designed to identify the main cell populations present in the sample –‘immunophenotypic differential’ - and to focus on particular populations of interest with respect to lineage assignment and differentiation stage. Aberrant antigen expression can be a key marker of abnormality. In addition detection of light chain restriction can be a very powerful tool for the analysis of clonality in B lymphoid populations.

Bone marrow and tissue samples are usually reported as a percentage of the population with the phenotype of interest. Peripheral blood usually includes absolute values.

Antibody panels

Based on the initial results other antibody combinations may be utilised. Immunocytochemistry can be performed on cell smears, when required.

 Specimen required

  • Blood (10ml EDTA) and/or
  • Bone marrow (1-2ml EDTA)
  • Bone marrow trephine (block or sections) for immunohistochemistry, if required
  • Fresh tissue biopsy or fine needle aspirate (FNA) in RPMI
  • Body fluids (in sterile tube) e.g. CSF, pleural fluid, Lavag

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

Sample storage  

Samples for flow cytometry should be analysed within 48 hours of collection. Samples should be kept at 4 degrees celcius 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. Scatter plots may be provided. An overall interpretation and a conclusion will be provided. Phenotypes, which may be useful for monitoring of minimal residual disease or response evaluation will be indicated.

Anticipated turnaround time

Results will be available within 48 hours of specimen collection, and urgent results can be obtained within a shorter time-frame if the laboratory is notified in advance.