Three-dimensional micro-PTV using deconvolution microscopy

Abstract

A three-dimensional micro-particle tracking velocimetry (micro-PTV) scheme is presented using a single camera with deconvolution microscopy. This method devises tracking of the line-of-sight (z) flow vectors by correlating the diffraction pattern ring size variations with the defocusing distances of small particle locations. The working principle is based on optical serial sectioning microscopy, or equivalently deconvolution microscopy, that records images of an infinitesimally small particle, and generates a point-spread function of the three-dimensional diffraction patterns. A new image-processing algorithm has also been developed to digitally identify the center locations and measure the radii of the diffraction rings, which allows simultaneous tracking of all three-vector components. The developed PTV technique uses a 40×, 0.75 NA dry objective lens with 500-nm fluorescent seeding particles of SG=1.05, and successfully measures the fully three-dimensional fields flowing over a spherical obstacle snuggly fitted inside a 100 μm × 100 μm micro-channel. The volumetric measurement resolution of the present system is equivalent to a 5.16 μm × 5.16 μm × 5.16 μm cube, and the overall measurement uncertainty for single-point velocity vector detection is estimated to ±7.58%.