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Three-dimensional micro-PTV using deconvolution microscopy

机译:使用反卷积显微镜的三维微型PTV

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摘要

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 40x, 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 mu m x 100 mu m micro-channel. The volumetric measurement resolution of the present system is equivalent to a 5.16 mu m x 5.16 mu m x 5.16 mu m cube, and the overall measurement uncertainty for single-point velocity vector detection is estimated to +/- 7.58%.
机译:使用具有解卷积显微镜的单个摄像机,提出了三维微粒跟踪测速(micro-PTV)方案。该方法通过将衍射图样环尺寸变化与小颗粒位置的散焦距离相关联,设计出对视线(z)流向量的跟踪。工作原理基于光学串行切片显微镜或等效的反卷积显微镜,可记录无限小的微粒的图像,并生成三维衍射图的点扩展函数。还开发了一种新的图像处理算法,以数字方式识别中心位置并测量衍射环的半径,从而可以同时跟踪所有三个矢量分量。先进的PTV技术使用40倍,0.75 NA干燥物镜和SG = 1.05的500-nm荧光播种粒子,并成功测量了流过紧密安装在100μmx 100μm微米内的球形障碍物上的全三维场-渠道。本系统的体积测量分辨率等于5.16μm×5.16μm×5.16μm立方体,并且单点速度矢量检测的总体测量不确定度估计为+/- 7.58%。

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