Flow cytometry

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Flow cytometry is an automated technique used to examine cells that is widely used for the diagnosis and characterization of lymphoma.

Principles

  1. Label cells with fluorescent antibodies.
  2. Passed through a channel one-at-a-time (using hydrodynamic focusing[1]) and interrogated with a laser light.

Scatter of light

Types:

  1. Forward scatter (FS) -- indicative of cell size.
  2. Side scatter (SS) -- indicative of cell complexity (granularity).

Scatter in a table

Selected white blood cells & scatter:[2]

Forward scatter Side scatter
PMNs high very high
Monocytes high moderate/low
Lymphocytes low low

Practical considerations

  • Tissue must be fresh, i.e. cannot be formalin fixed.
    • Viable material may be salvaged from the centre of a large specimen placed in formalin for a short time.
  • 2 mm x 2 mm x 2 mm piece of tissue is sufficient.
    • 1 mm3 is considered the minimum by Mayo Medical Laboratories.[3]

Interpretation

See Cytometry.

See also

References

External links