One hundred microliters of samples (i.e., conditioned culture medium or 10-fold diluted plasma) and 30 μl of diluted SERS nanotags were subsequently run in each microfluidic channel for 40 min under the field condition of 100 mV and 1 kHz. Under these previously optimized conditions (20), the stimulated nanoscopic fluid flow was the most efficient for EV capture and labeling due to the increased collision frequency of EVs with capture antibodies and SERS nanotags, and the minimum nonspecific binding.

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