A novel concept for continuous biosensing is introduced. Continuous biosensing is achieved by the mechanism named "Particle Cross Over" within microfluidic channels. The functionality of the device is demonstrated by quantifying the binding events of biotin to streptavidin coated beads. The device is designed to have precisely controlled flow structures which allow cytometric beads to pass from one flow region to another without mixing the carrier and sample fluids. The mean and standard deviation of the cytometric bead pixel intensities are 43.52 and 19.39, respectively for 1 μg/ml of biotinylated Fluorescein IsoThioCyanate concentration with a binding time of 0.464 seconds and shows a Gaussian distribution. It is expected that the particle cross over mechanism introduced in this report will be very useful mechanism for applications in continuous biosensing.

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