FIGURE 6.

Accurate characterisation of Extracellular Vesicles subpopulations at the single‐vesicle and single‐molecule level is dependent on the technical approach used for vesicles analysis. (a) Correlation analysis of GFP levels in small EVs derived from engineered Expi293F cells, as determined by Single‐Molecule Localization Microscopy (SMLM) versus Western blotting (left panel), SMLM versus ExoView (middle panel) and SMLM versus NanoAnalyzer N30 Nanoflow cytometry (right panel). For the ExoView dataset, the average of GFP MFI measured on the anti‐CD63, ‐CD81 and ‐CD9 spots was considered. Labels in the graphs indicate the sorting protein used for EVs engineering. All datasets were normalized to the untransfected control condition. R² values for the Pearson correlation between SMLM and the different analysis techniques are also indicated in the graphs