Flow cytometry is an automated analysis of cells passing in the fluid suspension through a laser light beam, which react with monoclonal antibodies specific for a variety of cell surface antigens. A specimen of peripheral blood, bone marrow, or other cell suspension is incubated with fluorescent-labeled antibodies, which bind to target antigens on cell surfaces or—following cell permeabilization—to cytoplasmic and nuclear antigens. The analysis of surface antigens is performed on cells selected (gated) based on light-scatter properties. The expression of specific marker or confirmation of markers defines a specific cell population or the original of these cells. This in turn helps in diagnosis and classification of various hematological diseases and leads to choosing a specific therapy. Here, we describe a methodology for using flow cytometry with six colors for the analysis of various tissues for hematological diseases.
Key Words: Flow cytometry; leukemia; lymphoma; immunophenotyping; methodology.
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