Flow cytometry immunophenotyping

Abstract:
Flow cytometry immunophenotyping is an important component in the diagnosis and evaluation of the course of  different diseases (see list below).

The method is based on the combination of flow cytometry and immunophenotyping with monoclonal antibodies. In flow cytometry, single cells of a suspension are analyzed by hydrodynamic focusing. At the measuring point, laser light strikes the cell and gives information on the size and inner structure of this cell via the scattering of light. Immunophenotyping is based on fluorochrome-tagged monoclonal antibodies against superficial, cytoplasmatic or intranuclear antigens. Modern, routinely used flow cytometry allows for the simultaneous examination of up to 10 differently tagged antibodies. The evaluation of 10'000 to over 1'000'000 analyzed cells is performed by a special software. Via electronic gating, even small cell populations can be looked at individually and be characterized. By examining high numbers of cells, the method can be used to detect minimal residual disease (MRD) under therapy. A sensitivity of 10^-5 provides comparable results as the PCR-method.

Indications in hematology:

• Diagnosis of acute myelocytic leukemia
• Diagnosis of acute lymphocytic leukemia
• Diagnosis of chronic lymphocytic leukemia
• Diagnosis of hairy cell leukemia
• Diagnosis of other mature B- and T-lymphocytic neoplasms
• Follow-up under therapy in hematological neoplasms including MRD-diagnosis (MRD = minimal residual disease)
• Quantification of blasts in myelodysplastic syndromes
• Diagnosis of myelodysplastic syndromes
• Mastocytosis
• Paroxysmal nocturnal hemoglobinuria
• Diagnosis of disorders of platelet function (Glanzmann's thrombasthenia , Bernard-Soulier syndrome)
• Quantification of CD34-positive stem cells for hematopoietic stem cell transplantation
• Diagnosis of hereditary spherocytosis (Eosin-5-Maleimid-binding-test)