Fig. 4.

Cell change by the different cell isolation techniques prior to flow cytometry analysis. a Lymphocytes (LYM) and monocytes (MONO) were equally affected by the two steps of density gradient centrifugation (DGC): DGC with 1300 rpm for 30 min (DGCun) and an additional washing step with 1300 rpm for 10 min (DGCw). During red blood cell lysis (RBCL), the amount of LYM lost was similar to both DGC steps, but cell gain in MONO seemed to be an artifact due to limitations of the hemocytometer used in this study. Data are displayed as geometric mean. n = 6. Repeated-measures ANOVA with Bonferroni post hoc comparisons was used for analysis. Significant losses to whole blood and differences between losses per cell type are indicated as follows: *p < 0.05, **p < 0.01, ***p < 0.001, and ^§§§p < 0.001, respectively. b Change of the lymphocyte-to-monocyte ratio between the different cell isolation steps. Individual values are displayed. n = 6. Repeated-measures ANOVA with Bonferroni post hoc comparisons was used for analysis. Significant differences are indicated as follows: *p < 0.05, **p < 0.01, ***p < 0.001