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Fig. 4 | Biological Procedures Online

Fig. 4

From: Improved cytometric analysis of untouched lung leukocytes by enzymatic liquefaction of sputum samples

Fig. 4

Cytometric gating strategy for the evaluation of frequencies and activation status of sputum leukocytes. Singlets (A), viable cells (B), total leukocytes (C) and total granulocytes/lymphocytes (D) are sequentially selected and then total lymphocytes and granulocytes are plotted in a SSC-A/CD11b-FITC dot-plot (E). Next, total granulocytes (red gate in E) are plotted in a CD16-PCy5/CD11b-FITC dot-plot and gates for selecting CD16High mature neutrophils and CD16Low neutrophils + eosinophils are defined (F). Finally, CD16Low neutrophils + eosinophils (red gate in F) are plotted in a SSC-A/FSC-A dot-plot and SSCHigh eosinophils are selected (G). Cell frequencies and the MFI of membrane markers (percentages and histograms, respectively, in panels E to G) are evaluated in total lymphocytes (pink) mature neutrophils (green) and eosinophils (blue) gates

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