Fig. 1

^EVHB-Chip design and reporter constructs. a Image of the operating COC ^EVHB-Chip. Pressure-driven flow-pumped serum or plasma through an inlet at 1 mL h⁻¹. Waste serum or plasma is collected in a single outlet (scale bar 1 cm). On the right, a SEM micrograph of the 3D herringbone features of the microfluidic device (scale bar 100 μm). b Gli36wt or Gli36-EGFRvIII glioma cells were stably infected with lentivectors encoding palmitoylated-tdTomato (PalmtdTomato) or -GFP (PalmGFP), respectively, to produce fluorescently labeled EVs. c Serum or plasma from healthy donors with spiked-in fluorescent EVs (in red and green) was run through the microfluidic device coated with tumor-specific antibodies to capture tumor EVs. d A collage of one hundred confocal microscopy images of fluorescent EVs captured on the surface of the device. Different antibodies were used: large image (Cetuximab, scale bar 50 μm); right insert (antibody cocktail); left insert (anti-EGFRvIII). e Representative image of digitally rendered signals of captured EVs shown as green (EGFRvIIIPalmGFP EV) or red spheres (WT-PalmtdTomato EV; scale bar 1 μm). f Quantification of captured wild-type PalmtdTomato and EGFRvIII PalmGFP EVs on the microfluidic device when different antibodies were used. A concentration of 20 μg mL⁻¹ were used for EGFRvIII and Cetuximab, and 10 μg mL⁻¹ for each antibody present in the cocktail (n = 3 technical replicates; ±standard error of the mean, s.e.m.). g After imaging captured EVs were lysed. Total RNA was isolated and correlated to confocal microscopy data (n = 3 technical replicates; ±s.e.m.)