STANOVENÍ INTRACELULÁRNÍCH MOLEKUL BUNĚKKREVNÍCH MALIGNIT PRŮTOKOVOU CYTOMETRIÍ

Detection of the Intracellular Molecules in Malignant Cells of HematopoieticOrigin by Flow Cytometry

Background.
Detection of membrane molecules by immunophenotypisation is a routine counterpart of thelaboratory tests which are needed for the complex diagnostic procedures of hematopoetic malignancies at present.The immunochemical detection of intracellular molecules is essential in such cases, where the expression of surfacemolecules on malignant cells is poor, or where the results of other diagnostic analyses are unequivocal (cases withectopic expression of other lineage-specific molecules and cases of biphenotypic leukemias).Methods and Results. We followed the procedure using 4% solution of paraformaldehyd in phosphate bufferedsaline for fixation of cells. Lysing solution G (Becton-Dickinson) was used for permeabilisation of cells to detectnuclear and cytoplasmatic molecules. The permeabilised cell suspension was than washed with 0,5% solution ofTween 20 in phosphate buffered saline. This simple and rapid procedure is readily available for the routine detectionof intracellular molecules. This approach is important for the diagnosis of hematopoetic malignancies.

Key words:
immunophenotypisation, flow cytometry, cytoplasmatic molecules, nuclear molecules.