lymphoma. The reproducibility of the data (scatter plots and histograms) further enhances the utility of the information in the context of disease monitoring and tracking of residual disease in hematolymphoid malignancies. The combined power of these two major components of the information significantly and exponentially enhances the diagnostic potentials of multicolour immunophenotyping. As mentioned above, multicolour flow cytometry can generate maximum amount of information from the smallest possible tissue sample. By using multiple anti- body reagents directed against different antigens, a large body of information could be obtained about the antigenic profile of cells from a single assay tube. There- fore, this approach is ideally suited for examining precious and small samples, e.g. CSF, FNA, bone marrow and lymph node biopsy as it obviates the need to put up multiple assay tubes. The savings on the cost of the rea- gents, consumables, manpower, equipment and infrastructure on account of use of less number of assay tubes per sample in multicolour flow cytometry could be enormous in the long run. With rising cost of healthcare, this advantage could be very attractive.
“It is quite apparent that most advanced clinical flow cytometry laboratories have settled for 8-10 colour analysis after a lot of trial and error”
The use of less number of assay tubes in multicolour flow cytometry makes it possible to see and acquire a larger number of cells per tube. This in turn allows clear visualization and analysis of small populations of cells of interest which are not clearly visible when one acquires a smaller number of total cells. Selection and standardization of antibody panels for diagnosis of hematol- ymphoid malignancies is a major chal- lenge since consensus among clinical flow cytometry laboratories on the number, choice and combination of various antibodies and fluorochromes for the diagnosis of these disorders has not been reached. The situation gets more compli- cated when the laboratory uses only 2-4 colour combinations. The choice of the
best possible combination of antibodies is a matter of debate under these circum- stances. The use of larger multicolour rea- gent combinations (panels) that incorpo- rate commonly used antibodies and those of diagnostic (and prognostic) importance circumvents this issue to a large extent.
SOME SPECIFIC CONTRIBUTIONS OF MULTICOLOUR FLOW CYTOMETRY The information from flow cytometric immunophenotyping of hemopoietic cells in myelodysplastic syndromes (MDS) available thus far strongly points to a variety of changes in the surface antigenic composition of these cells which deviate significantly from the normal antigenic profiles of cells at different stages of maturation and follow a predictable pattern in many instances. These changes are not seen in conditions which are associated with morphologic features similar to those seen in MDS but don’t represent a preleukemic state. The ability to use 8-10 or more reagents in each tube increases the power of the technology and allows distinction between these two sets of conditions with greater ease and clarity. Detection and monitoring of minimal residual disease has been in practice for
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