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. 2023 Mar 5;13(5):1470–1489. doi: 10.7150/thno.77597

Figure 3.

Figure 3

Non-conventional flow cytometry strategy used to characterize sEVs. (A) CFDA-SE staining for identification of intact vesicles. Red areas identify CFDA-SE positive events. EVs were stained with CFDA-SE at 4 °C as "blank tube" (left), useful to define the appropriate dimensional gate when considering EVs stained with CFDA-SE at room temperature (right). (B) Size distribution of EV subtypes. Three dimensional gates were considered: EVs ≤100 nm (orange), 100 nm ≤ EVs ≤ 160 nm (blue) and 160 nm ≤ EVs ≤ 900 nm (red). (C) Quantification of CD9-, CD63- and CD81- positive events falling within the CFDA-SE gate. Data are presented as ratio between mean fluorescence intensity (MFI) of sEVs stained with a specific antibody and MFI of correspondent isotype control (relative MFI). Data are representative of at least three independent experiments.