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Proceedings Paper

Three-Dimensional Microscopy: Image Processing For High Resolution Subcellular Imaging
Author(s): David A. Agard; Yasushi Hiraoka; John W. Sedat
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Paper Abstract

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.

Paper Details

Date Published: 22 December 1989
PDF: 7 pages
Proc. SPIE 1161, New Methods in Microscopy and Low Light Imaging, (22 December 1989); doi: 10.1117/12.962684
Show Author Affiliations
David A. Agard, University of California at San Francisco (United States)
Yasushi Hiraoka, University of California at San Francisco (United States)
John W. Sedat, University of California at San Francisco (United States)


Published in SPIE Proceedings Vol. 1161:
New Methods in Microscopy and Low Light Imaging
John E. Wampler, Editor(s)

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