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29 March 2005 Improved 3D cellular imaging by multispectral focus assessment
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Biological specimens are three-dimensional structures. However, when capturing their images through a microscope, there is only one plane in the field of view that is in focus, and out-of-focus portions of the specimen affect image quality in the in-focus plane. It is well-established that the microscope’s point spread function (PSF) can be used for blur quantitation, for the restoration of real images. However, this is an ill-posed problem, with no unique solution and with high computational complexity. In this work, instead of estimating and using the PSF, we studied focus quantitation in multi-spectral image sets. A gradient map we designed was used to evaluate the sharpness degree of each pixel, in order to identify blurred areas not to be considered. Experiments with realistic multi-spectral Pap smear images showed that measurement of their sharp gradients can provide depth information roughly comparable to human perception (through a microscope), while avoiding PSF estimation. Spectrum and morphometrics-based statistical analysis for abnormal cell detection can then be implemented in an image database where the axial structure has been refined.
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Tong Zhao, Yizhi Xiong, Alice P. Chung, Elliot S. Wachman, and Daniel L. Farkas "Improved 3D cellular imaging by multispectral focus assessment", Proc. SPIE 5699, Imaging, Manipulation, and Analysis of Biomolecules and Cells: Fundamentals and Applications III, (29 March 2005);

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