Flow cytometers
A machine that streams cells one at a time past lasers and reads the fluorescent tags stuck to each one, counting tens of thousands of cells a second. It is how leukaemias and lymphomas are typed, how residual disease is measured after treatment, and how cell therapies are checked before infusion.
Overview
In a flow cytometer, cells labelled with fluorescent antibodies are focused into a single-file stream and cross one or more laser beams; detectors record the scattered light (size and granularity) and the fluorescence from each dye, so every cell is scored for a dozen or more surface and internal markers at once. Haematology laboratories use it to assign lineage in acute leukaemia, subtype lymphomas, count CD34 stem cells in a transplant harvest and, with standardised panels such as EuroFlow, detect measurable residual disease down to one abnormal cell in ten thousand or fewer in acute lymphoblastic leukaemia, myeloma and chronic lymphocytic leukaemia. Cell therapy manufacturers use it to characterise CAR-T products. Conventional instruments (BD FACSLyric and FACSCanto, Beckman Coulter Navios EX and DxFLEX) run eight to twelve colours; full-spectrum cytometers (Cytek Aurora, Sony ID7000) resolve forty or more; cell sorters divert chosen cells into tubes for culture or sequencing; and mass cytometry replaces fluorophores with metal tags read by a mass spectrometer.
The method needs fresh, live cells within hours of sampling, skilled gating and standardised panels for results to be comparable between laboratories, and sequencing-based residual disease tests are more sensitive for some diseases.
- Plasma
- Tumour-informed variants at < 0.01%
- Serial draws after surgery
How it works
Hydrodynamically focused cells pass single file through laser beams; scattered light and multicolour fluorescence from antibody-bound dyes are recorded per cell, so populations are enumerated and phenotyped by marker combinations.
- Tens of thousands of cells a second, many markers per cell
- Standard for leukaemia typing and flow-based residual disease
- Sorts live cells for downstream assays
- Needs fresh viable cells within hours
- Gating is operator dependent without standardised panels
- Less sensitive than sequencing for some residual disease tests
Latest papers
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