Figure 15-2. Selection of leukemic blast cells by flow cytometry. Cells of interest are selected based on particular criteria. Blasts typically express low side-scatter, dim CD45, and/or specific surface markers (e.g., CD34). This figure demonstrates how gating on the specific marker selects the precise population, which otherwise might be difficult to discern from neighboring cell clusters, as pictured in the "R1 alone" plot.

For body fluids other than peripheral blood or bone marrow, such as peritoneal or pleural effusions or cerebrospinal fluid, addition of anticoagulants is not necessary. Biopsies from lymph nodes, skin, or soft tissue tumors should be collected in tissue culture medium (e.g., RPMI 1640), and cell suspensions should be prepared by mechanical disruption of the tissue. The viability of the white cells in a specimen should be checked to ensure adequate quality of the sample.

Immunohistochemical evaluation of slide preparations (e.g., bone marrow core biopsies or granulocytic sarcoma sections) in acute leukemia is less informative if a distinction between normal and leukemic cells cannot be made and/or the cellular localization of antigens is equivocal because of inferior morphology after freezing or fixation.95,96 In contrast to the situation with lymphomas, preserving the tissue architecture is of no concern in the immunophenotypic diagnosis of the acute leukemias. Furthermore, if formaldehyde-fixed sections are tested, fewer antibodies are available because of the fixation.97

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