Optimizing Extracellular Vesicle Biotinylation Analysis
Corresponding Organization : Nankai University
Variable analysis
- Optimization of electroporation
- Biotinylation efficiency of EVs
- The use of an Apogee A‐50 Micro Flow Cytometer (Apogee Flow Systems) equipped with 375, 405, 488, and 638 nm lasers
- The use of reference beads (ApogeeMix, Apogee Flow Systems) composed of a mixture of nonfluorescent silica beads (Si, refractive index ≈1.42) with diameters of 180, 240, 300, 590, 880, 1300, and 110 nm, 500 nm green fluorescent latex spheres (polystyrene, refractive index ≈1.59) to set the thresholds for light scattering and help to gate sEVs and lEVs
- The tubing was washed with double‐distilled water after each sampling
- Analysis was performed at a flow rate of 1.5 µL min^-1 using a 150 µL sample volume for at least total 3 × 10^5 particles
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