Simple and low-cost implementation of three-dimensional (3D) particle measurement is vital for designing and characterizing microfluidic devices that show spatiotemporally varying characteristics in three dimensions. However, the conventional 3D particle image velocimetry or particle streak velocimetry has proven difficult to address the needs, requiring complex and expensive equipment, precise alignment between optical components, and specialized image-processing algorithms. Here, we report mirror-embedded microchannels and a method of optical path-length (OPL) modulation for 3D particle measurement in the channels. The mirror, ideally at 45°, reflects the side view of the channels and enables 3D positional information to be obtained easily from two different orthogonal-axis images with different optical paths. To offset the optical path difference between two image views, we utilized a cover glass as a medium of high refractive index and placed it in the light path through which the side-view image propagates, thereby prolonging the OPL of the side view and simultaneously shifting its depth of field (DOF) range. This modulation ensures imaging of in-focus side view as well as top view. This 3D imaging principle was verified by observing 3D positions of 6 μm-sized beads in the linear and grooved microchannels. The mirror-embedded scheme can be readily fabricated with existing microfluidic designs, and offer easy and simple implementation of 3D particle measurement.
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