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Light Microscopy Image Collection: Confocal, Widefield and Deconvolution

Published online by Cambridge University Press:  02 July 2020

J. N. Turner ,
D. H. Szarowski ,
B. Roysam  and
T. J. Holmes
J. N. Turner
Affiliation:
Wadsworth Center, New York State Department of Health, Albany, NY, 12201 Rensselaer Polytechnic Institute, Troy, NY, 12180
D. H. Szarowski
Affiliation:
Wadsworth Center, New York State Department of Health, Albany, NY, 12201
B. Roysam
Affiliation:
Rensselaer Polytechnic Institute, Troy, NY, 12180 AutoQuant Imaging Inc., Troy, NY, 12180
T. J. Holmes
Affiliation:
Rensselaer Polytechnic Institute, Troy, NY, 12180 AutoQuant Imaging Inc., Troy, NY, 12180
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Extract

Light microscopy instrumentation and biological applications continue to expand rapidly, and an important aspect of this expansion is three-dimensional (3-D) imaging and image analysis. For many biological specimens, optical sectioning, i.e. collecting images at a sequence of depths under controlled conditions, provides true 3-D data through the entire specimen. This is especially important for specimens whose thickness exceeds the depth-of-field of the microscope objective lens. This sequence of optical sections is the basis for 3-D image reconstruction providing information from all three specimen dimensions instead of just the traditional two in the image plane of the microscope. The analysis of images in 3-D provides insights into the structure and function of biological specimens that are not available through other means. However, 3-D microscopy also presents additional choices for image collection. The first of which is whether to use widefield or confocal microscopy and the second is whether to utilize digital deblurring or deconvolution methods.

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Tutorials
Information
Microscopy and Microanalysis , Volume 3 , Issue S2: Proceedings: Microscopy & Microanalysis '97, Microscopy Society of America 55th Annual Meeting, Microbeam Analysis Society 31st Annual Meeting, Histochemical Society 48th Annual Meeting, Cleveland, Ohio, August 10-14, 1997 , August 1997 , pp. 371 - 372
Copyright
Copyright © Microscopy Society of America 1997

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7. Support: NIH R43-EY11174, R43-MH53691, R43-MH53692; NSF BES-9413037.Google Scholar