22 December 1989 Three-Dimensional Microscopy: Image Processing For High Resolution Subcellular Imaging
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Recent technological advances now make it practical to record three-dimensional data from biological specimens using fluorescence light microscopy. When three-dimensional images are collected using a conventional microscope, each observed section contains in-focus information from the parts of the sample at the focal plane and out-of-focus information from the remainder of the sample. The imaging process can be characterized as a convolution of the sample with the point spread function (PSF) of the microscope. We have experimentally determined the PSF for an epifluorescence microscope using high numerical aperture oil and water immersion lenses. Several methods for the processing of the observed data are discussed, with the best results obtained by constrainediterative deconvolution methods.
© (1989) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
David A. Agard, David A. Agard, Yasushi Hiraoka, Yasushi Hiraoka, John W. Sedat, John W. Sedat, } "Three-Dimensional Microscopy: Image Processing For High Resolution Subcellular Imaging", Proc. SPIE 1161, New Methods in Microscopy and Low Light Imaging, (22 December 1989); doi: 10.1117/12.962684; https://doi.org/10.1117/12.962684


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