1 

122 Flow Cytometry - Select Topics

B 

and 351 (mut).

IPS = 0 

CD33 

CD11b 

CD16 

CD13 

CD45 

CD15 

CD64 

MPO 

**Figure 2. NPM1 mutational analysis on sorted cell fractions.** Cell compartments are shown at the top; as concerns neutrophil compartment (left), the results of clustering analysis are depicted, together with phenotypic parameters and compartment's phenotypic score (IPS), appraising the extent of dysplasia. In the corresponding plots, the cell popula‐ tion is highlighted by color: blue for neutrophil cells, red for blasts, orange for T lymphocytes. The relative data from NPM1 mutational analysis are reported below. (A) Patient #1: neutrophil compartment showed several phenotypic abnormalities (as represented by clustering analysis and IPS = 6.0) and harbored NPM1 mutation; sorted blasts and T lymphocytes were NPM1-mutated and wild-type, respectively. (B) Patient #2: neutrophil compartment showed a pre‐ served phenotypic profile (IPS = 0) and a NPM1 wild-type status. Sorted blasts and T lymphocytes were NPM1-mutat‐ ed and wild-type, respectively. Clustering analysis was performed by R software. Dot plots were created by Infinicyt software. NPM1 mutational analysis: in order to discriminate NPM1 PCR products, we used 5'- end HEX dye-labeled reverse primer (M-Medical). The amplified products were separated with a capillary electrophoresis-based system us‐ ing ABI PRISM 310 genetic analyzer (Applied Biosystems). The labeled fragment size corresponding to NPM1 wildtype gene was 347 bp. All NPM1-mutated samples were heterozygous, showing a double peak at positions 347 (wt)

CD64% 

FSC 

SSC 
