Implementation of flow cytometry in the diagnostic work-up of myelodysplastic syndromes in a multicenter approach: Report from the Dutch Working Party on Flow Cytometry in MDS

Standardization of flow cytometry in myelodysplastic syndromes: a report from an international consortium and the European LeukemiaNet Working Group

“…12 CD33 and CD64 are useful in distinguishing monocytes and hypogranular neutrophils. 31 Otherwise, a computer-assisted multidimensional, instead of simply multiparameter, analysis might overcome problems encountered when subpopulations overlap.…”

“…As noted above, maturing neutrophils with an abnormally low SSC can interfere in the analysis of the monocytic population, depending on the gating strategies. CD33 or the combination of CD64 and CD24 might be useful to separate these two subpopulations (examples in Westers et al 31 ). Of note, doublets of cells from both populations might also hamper analysis and should be excluded in an FSC-height versus FSC-area dot plot.…”

Immunophenotypic analysis of erythroid dysplasia in myelodysplastic syndromes. A report from the IMDSFlow working group

“…Examples of antibody combinations and panels have been described previously. 3,26,27 Nuclear dyes were not routinely included in the panels, and only one center applied the live/dead stain 7-AAD. The flow cytometers used included: FACSCalibur (BD Biosciences; n=3); FACS CANTO-II (BD Biosciences; n=10); a combination of FACSCalibur and FACS CANTO-II (both BD Biosciences; n=2); and Navios (Beckman Coulter; n=4).…”

Loss of Blast Heterogeneity in Myelodysplastic Syndrome and Other Chronic Myeloid Neoplasms

“…17 There are two approaches in FCIP for myeloid neoplasms (as well as in other FCIP applications): ones that define abnormalities quantitatively using mean fluorescence intensity of individual antigens, 7,8,15,16,18,19 and ones that define abnormalities qualitatively, looking at the pattern of expression. 4,6,10,[20][21][22][23][24][25] These two approaches differ in test performance characteristics and validation requirements. 26 Quantitative approaches have the advantage of being more reproducible because of no interobserver variability, and they are easily automated using specialized software for data analysis 13 ; however, quantitative approaches rely on very stringent specimen and instrument settings and quality control, which is difficult to standardize among laboratories.…”

“…[3][4][5][6][7][8][9][10] However, FCIP for MDS also has many limitations preventing it from routine clinical use: (1) no single antigen expression is diagnostic, (2) different research groups use different sets of markers and/or reagents with poor interlaboratory reproducibility, (3) the clinical usefulness of FCIP is uncertain because most studies examine only specimens with morphologically and/or cytogenetically obvious dysplasia, and (4) analytic approaches vary from looking at differentiated myeloid cells to others that focus on blast populations. Most approaches for distinguishing normal from dyspoietic myeloid maturation use a large number of antibodies, combined with either a point-based system for defining the level of abnormality, 3,11 or a specialized analytic software for multidimensional data analysis.…”