An Imaging Flow Cytometry-Based Methodology for the Analysis of Single Extracellular Vesicles 3 75 Framework Criteria What to report Please complete each criterion 6.3 EV number/ concentration. State whether EV number/ concentration has been reported. If calculated, it is preferable to report EV number/ concentration in a standardized manner, stating the number/ concentration between a set detection range. Detected concentrations of fluorescent EV are described in detail in the manuscript. All concentrations reported were obtained between the calibrated detection ranges for each channel, as described above. 6.4 EV brightness. When applicable, state the method by which the brightness of EVs is reported in standardized units of scatter and/or fluorescence. EV brightness was calculated for all fluorescent populations analyzed and described in the work; MFI values were converted into standardized-ERF values. For CFSE+Tetraspanin+ EV measured in the PPP samples, we observed a mean EV brightness of 119.57 ERF (range 99.6-156) for CFSE and 65.33 ERF (range 61.369.8) for APC. For CD9+CD31+ EV measured in the PPP samples, we observed a mean EV brightness of ~7,620 (range 3,640 – 9,240) and 20.4 (range 15 – 27.9) for BV421 and APC, respectively. 7.1. Sharing of data to a public repository. Provide a link to the experimental data in a public data repository. IFCM files can be obtained by contacting the corresponding author. Supplementary Table 1 – Framework representing the Minimal Information about a Flow Cytometry (FC) experiment to allow standardized EV-FC-specific reporting (MIFlowCyt-EV template), as recommended by the Minimum Information for Studies of EVs (MISEV).
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