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The Particle Metrix ZetaView® instrument family can be equipped with a fluorescence function. Depending on the number and wavelength of the installed lasers, Brownian Motion Tracking and Electrophoretic Motion Tracking (EMT) / zeta potential of fluorescent particles can be recorded in 1, 2 or 4 fluorescence channels. In this video we show an example with extracellular vesicles stained with AlexaFluor® 488 and show how fluorescence measurements are performed by the Particle Metrix ZetaView® instruments. We show which settings are important and how they are to be carried out step by step.