CD63 and cancer: Considering that the CD63 and miR-21 positive EV populations were only 8.5% (Figure 7C) and 20.3% (Figure 7D) in clustered normal and un-clustered cancer EVs, respectively, we concluded that the novel method for single step in situ detection of protein and miRNA biomarkers in EVs using flow cytometry can be significantly improved upon clustering the EVs using DSPE-PEG-DSPE.