Three-dimensional three-component particle velocimetry for microscale flows using volumetric scanning

We present a diagnostic platform for measuring three-dimensional three-component (3D3C) velocity fields in microscopic volumes. The imaging system uses high-speed Nipkow spinning disk confocal microscopy. Confocal microscopy provides optical sectioning using pinhole spatial filtering which rejects light originating from out-of-focus objects. The system accomplishes volumetric scanning by rapid translation of the high numerical aperture objective using a piezo objective positioner. The motion of fluorescent microspheres is quantified using 3D3C super resolution particle-imaging velocimetry with instantaneous spatial resolutions of the order of 5 mu m or less in all three dimensions. We examine 3D3C flow in a PDMS microchannel with an expanding section at 3D acquisition rates of 30 Hz, and find strong agreement with a computational model. Equations from the PIV and PTV literature adapted for a scanning objective provide estimates of maximum measurable velocity. The technique allows for isosurface visualization of 3D particle motion and robust high spatial resolution velocity measurements without requiring a calibration step or reconstruction algorithms.