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A Robust Two-View Method for Increasing the Imaging Depth and Correcting for Signal Attenuation in Confocal Microscope Images

Published online by Cambridge University Press:  02 July 2020

Ali Can ,
Shade Lasek ,
Donald H. Szarowski ,
James N. Turner  and
Badrinath Roysam
Ali Can
Affiliation:
Rensselaer Polytechnic Institute, 110, 8, th Street, Troy, New York, 12180
Shade Lasek
Affiliation:
Wadsworth Center, NY State Department of Health, Albany, New York, 12201
Donald H. Szarowski
Affiliation:
Rensselaer Polytechnic Institute, 110, 8, th Street, Troy, New York, 12180
James N. Turner
Affiliation:
Rensselaer Polytechnic Institute, 110, 8, th Street, Troy, New York, 12180 Wadsworth Center, NY State Department of Health, Albany, New York, 12201
Badrinath Roysam
Affiliation:
Rensselaer Polytechnic Institute, 110, 8, th Street, Troy, New York, 12180
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Extract

Confocal Microscopy is now an essential, and widely used tool for biological research. Particularly valuable is its ability to image thick sections of intact tissue. A compelling need in this field is to improve the imaging depth, as well as quantitative accuracy, and minimize the possibility of missing important structures/phenomena due to attenuation of the imaging signal. Previous methods work by amplifying the signal from deeper parts of the specimen. In these methods, amplification of noise is an unavoidable artifact. We describe a novel method that not only corrects confocal stacks for attenuation without noise amplification, but also enhances the achievable imaging depth. Interestingly, it does not require explicit estimation of the extinction factor, It relies on a synergistic combination of specimen preparation and image reconstruction algorithms.

The key idea is to image the specimen from two angles (Fig. 1), separated by 180° and then to reconstruct the complete thick image using computational methods.

Type
Confocal Microscopy
Information
Microscopy and Microanalysis , Volume 6 , Issue S2: Proceedings: Microscopy & Microanalysis 2000, Microscopy Society of America 58th Annual Meeting, Microbeam Analysis Society 34th Annual Meeting, Microscopical Society of Canada/Societe de Microscopie de Canada 27th Annual Meeting, Philadelphia, Pennsylvania August 13-17, 2000 , August 2000 , pp. 818 - 819
Copyright
Copyright © Microscopy Society of America

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References

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