[Immunofluorescence and immunocytochemical stain methods for simultaneous cytogenetic and phenotypic characterization...

Cytogenetic analysis and immunological phenotyping are well established methods for characterizing leukaemic cell populations. The simultaneous evaluation of cell phenotype and karyotype has, however, been established only recently. We present immunocytochemical methods utilizing horseradish peroxidase and alkaline phosphatase monoclonal anti-alkaline phosphatase staining procedures, as well as modified immunoflurescence methods with fluorescein- and rhodamine-conjugated antibodies. Chromosomes were either stained with Giemsa and "stains-all" (immunocytochemical techniques) or chromomycin and distamycin A/DAPI for R- and C-banding (immunofluorescence techniques), respectively. The advantages and disadvantages of these methods are compared and provide representative examples of the various preparations. These methods will contribute to defining the phenotype of cytogenically-normal cells in leukaemias and to assigning different karyotypes to heterogenous cell populations, as well as supporting the search for analysis of residual blast cells.