Understanding the motion of colloidal particles flowing in small spaces is a general issue in various fields such as thermal engineering and micro/nanofluidics. In the present study, we investigated the motion of fluorescent submicrometer particles in a 3-μm microchannel by defocusing nanoparticle image velocimetry. An optical measurement system with controlled spherical aberration and an algorithm for processing defocused particle images with multiple diffraction rings were developed. By detecting the centroid position and the diameter of the outermost diffraction ring, which is proportional to the distance between the focal plane and the particle, the position of particles was determined with the spatial resolutions of 154–204 nm in the streamwise direction and 76–311 nm in the depthwise direction, which are comparable to or smaller than the optical diffraction limit. A reusable microfluidic device containing a size-regulated microchannel made of glass was developed, which is suitable for optical measurements and precise flow control. By controlling the strength of low-temperature glass bonding, detachment of the bonded glass substrates, washing, and reuse were achieved. Based on this method and technology, the velocity of particles with diameters of 199, 457, and 1114 nm was successfully measured in pressure-driven laminar flow. Results suggested that for larger particles comparable to the channel size, the particle velocity is slowed from the flow velocity by particle–wall hydrodynamic interactions. Therefore, the motion of colloidal particles in 100-μm spaces is considered to be affected by particle–wall hydrodynamic interactions, as well as 102-μm spaces reported previously.
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