Journal: PLoS ONE
Article Title: Analysis of cell surface and intranuclear markers on non-stimulated human PBMC using mass cytometry
Figure Lengend Snippet: Effects of different buffers on the detection of cell surface markers. Cells from healthy donors were labelled with antibodies targeting both cell surface and intranuclear antigens. Different permeabilization conditions are compared to the cell surface staining only” (CS) condition: ICSb (BD cytofix/cytoperm buffer), INSb 1 (eBioscience permeabilization buffer), INSb 2 (Maxpar NASB) and INSb 3 (Methanol/PFA). Here, we show the effects of different permeabilization conditions on the frequency of various cell surface markers. a.) Histograms showing the frequency and distribution of CD45+, CD19+, CD3+, CD16+, CD56+, CD14+ and HLADR+ events in the CS condition. The concentrations of antibodies used were: CD45 (0.5 mg/ml), CD19 (0.5 mg/ml), CD3 (0.08 mg/ml), CD16 (0.2 mg/ml), CD56 (0.1 mg/ml), CD14 (0.3 mg/ml) and HLADR (0.3 mg/ml). b.) Comparison of the frequency of CD45+, CD19+, CD3+, CD16+, CD56+, CD14+ and HLADR+ events between the CS condition and the different permeabilization conditions. Statistics was performed using one-way ANOVA with Bonferroni’s multiple test correction (*p
Article Snippet: Fixation and permeabilization using BD cytofix/cytoperm buffer (BD, France, Catalog # 554715) Cells were incubated at RT for 1 hour with BD Cytofix/Cytoperm buffer, then washed twice with BD wash/perm solution diluted 10X in distilled water.