Journal: Journal of Extracellular Vesicles
Article Title: A subset of extracellular vesicles carries the bulk of microRNAs in commercial dairy cow’s milk
Figure Lengend Snippet: High-sensitivity flow-cytometry analysis of fractionated IDG revealed the peculiar enrichment of EVs in the 12,000 g pellet. (a) A pool of all the IDG fractions was labelled with deep red CellTracker fluorescent dye and the gates set for fluorescent EV (green) detection, compared with unlabelled EVs (Red) and CellTracker in PBS-loaded (empty) IDG fractions (PBS, Blue). (b–c) Each IDG fraction of the 12,000 g (b) and 35,000 g (c) was labelled and analysed for the presence of esterase-positive EVs. Results are expressed as a count of events (mean ± SD; n = 3). (d) After setting the gates for size, using beads of known diameter, we analysed the CellTracker positive population in each fraction to determine the approximate size of the observed events. (e–f) Representative results obtained for IDG fractions F7 and F8 from 12,000 g (e) and 35,000 g (f) EVs are displayed. A green colour represents CellTracker positive events. The associated quantification results are available as Supplementary File 5.
Article Snippet: To this end, 2 µL of each IDG fraction was diluted in 98 µL of 0.2 µm filtered sterile PBS (Gibco, ThermoFisher scientific) containing the fluorescent CellTracker™ deep red dye [ ] (C34565; Life Technologies, ThermoFisher Scientific) at a final concentration of 1 µM, for 20 min at 37°C, in the dark, following the manufacturer’s recommendations.
Techniques: Flow Cytometry, Cytometry