fig2

Figure 2. The selected anti-CD9, anti-CD63, anti-CD66b, anti-CD82, and anti-HLA-DR antibodies enable labeling of detergent-sensitive objects in marginally processed synovial fluid. For the evaluation of background signals, antibodies were applied to PBS (1st row); for the detection of EVs, aliquots of a synovial fluid sample that had been preprocessed by 2,000 and 10,000 × g centrifugation were labeled with each of the five different antibodies (2nd row). To approve the EV nature of identifiable objects, detergent controls were performed; specifically, NP40 was added to labeled synovial samples before analysis (3rd row). Samples were analyzed by imaging flow cytometry using the gating strategy as depicted in Figure 1A. Drawing Software: CorelDraw. EV: Extracellular vesicle.