Hi, This is protocol for staining EVs from mouse plasma. You can use it for staining EVs in the cell's media too:

C57BL/6 mouse plasma is thawed, centrifuged for 30 min at 3000 g, and submitted to microfiltration with 0.22-mm filters to remove cell debris and apoptotic bodies. Filtered plasma is stained for 20 min at 37°C in the dark with 1 µM of DiL dye (Thermo Fisher Scientific, Waltham, MA, USA). Immediately after staining, EVs are extracted from plasma samples by size exclusion chromatography (SEC) method using the qEV columns (Izon Science, Christchurch, New Zealand). For more details see our paper : PMID: 39554866.

Thanks a lot for your answer @Vasily