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. 2018 Jun 13;9:1326. doi: 10.3389/fimmu.2018.01326

Figure 1.

Figure 1

Multiplex bead-based flow cytometry assay principle for detection of extracellular vesicle (EV) surface signatures. (A) Overview of assay workflow. 39 multiplexed populations of dye-labeled antibody-coated capture beads are incubated with EV-containing samples. In this case, captured EVs are counterstained with APC-labeled detection antibodies by using a mixture of anti-CD9, anti-CD63, and anti-CD81 (pan tetraspanin) antibodies. (B) Results after analyzing HEK293T conditioned medium (CM) compared to the respective medium control, showing all 39 bead populations identified by their fluorescence in the FITC Vs. PE channel (see Table S1 in Supplementary Material) with adjunct dot plots showing respective APC-stained bead populations. Back-gating from the four bead populations with the brightest staining (CD9, CD29, CD63, and CD81) is shown as an example to underline the assay principle (bottom right). (C) Representative quantification of the median APC fluorescence values for all bead populations after background correction (medium control values subtracted from measured HEK293T CM values). See Figures S1 and S2 and Tables S1 and S2 in Supplementary Material for further details.