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Immunophenotyping (Flow Cytometry)

• Immunophenotyping team

• About our flow cytometry service

• Specimen requirements

• Antibody panels

Scientist in Charge

• David Bloxham (01223 586546)

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About our flow cytometry service

We use flow cytometry to analyse cell populations in detail. Our current process includes:

• 8-colour panels (with plans to introduce 12-colour panels)

• forward and side scatter properties of cells

Our antibody panels help us:

• identify main cell populations in samples

• focus on specific populations to determine:

  • lineage assignment

  • stage of differentiation

We look for signs of abnormality, including:

• aberrant antigen expression

• asynchronous antigen expression

• light chain restriction in B-cell populations (helps detect clonal B-lymphoid populations)

How we report results:

• For bone marrow and tissue samples:

  • percentage of populations of interest

  • phenotype description

• For peripheral blood lymphoid populations:

  • typically includes absolute values

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Specimens required for flow cytometry

We accept the following specimens:

Blood (10 mL EDTA) and/or bone marrow (1–2 mL EDTA)

• Include an unstained or stained smear preparation for evaluation

• This can be the same sample provided for morphology

Bone marrow trephines (in sterile saline or RPMI)*

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

Fresh tissue biopsy* or fine needle aspirate (FNA) (1 mL sterile saline or RPMI)

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

Apheresis collection and/or cord blood (in EDTA or ACD)

Important

*For bone marrow trephine or tissue biopsy, always place the first obtained sample in formalin for primary haematopathology reporting. Use flow cytometry only as a secondary investigation.

Sample preparation and storage

To ensure reliable results:

• Analyse samples within 48 hours of collection

• Store samples at 4°C before despatch

• Process fresh tissue biopsies and body fluids as soon as possible after collection

These steps help preserve marker expression for immunophenotyping and maximise cell viability.

Important

Delayed processing may lead to unreliable or uninterpretable results.

Please phone the laboratory in advance when fresh tissue biopsies or body fluid samples are expected.

Flow cytometry reporting

Our immunophenotyping reports include:

• Clinical details from the request form

• Sample type and cell population analysed

• Percentage of cells with the phenotype of interest, relative to the whole population

• Overall interpretation and conclusion

• Phenotypes useful for monitoring minimal residual disease or evaluating response

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Routine antibody panels

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