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High-resolution restoration of 3D structures from widefield images with extreme low signal-to-noise-ratio

High-resolution restoration of 3D structures from widefield images with extreme low signal-to-noise-ratio
High-resolution restoration of 3D structures from widefield images with extreme low signal-to-noise-ratio

Four-dimensional fluorescence microscopy--which records 3D image information as a function of time--provides an unbiased way of tracking dynamic behavior of subcellular components in living samples and capturing key events in complex macromolecular processes. Unfortunately, the combination of phototoxicity and photobleaching can severely limit the density or duration of sampling, thereby limiting the biological information that can be obtained. Although widefield microscopy provides a very light-efficient way of imaging, obtaining high-quality reconstructions requires deconvolution to remove optical aberrations. Unfortunately, most deconvolution methods perform very poorly at low signal-to-noise ratios, thereby requiring moderate photon doses to obtain acceptable resolution. We present a unique deconvolution method that combines an entropy-based regularization function with kernels that can exploit general spatial characteristics of the fluorescence image to push the required dose to extreme low levels, resulting in an enabling technology for high-resolution in vivo biological imaging.

Algorithms, Animals, Cell Line, Entropy, Imaging, Three-Dimensional/methods, Microscopy, Fluorescence/methods, Models, Molecular, Models, Theoretical, Nuclear Proteins/chemistry, Protein Conformation, Reproducibility of Results, Saccharomyces cerevisiae/cytology, Saccharomyces cerevisiae Proteins/chemistry, Signal-To-Noise Ratio
0027-8424
17344-17349
Arigovindan, Muthuvel
3b1f076d-191e-4268-9283-689e48b77a33
Fung, Jennifer C.
8617f84c-7e4a-4d1b-898c-b51a600d46d9
Elnatan, Daniel
1065ca8c-5c21-4144-b131-a33417312e02
Mennella, Vito
43c60e29-c0a7-4ab8-8e5c-fcb59f70a28a
Chan, Yee-Hung Mark
9068ef12-a2cf-4d5b-bc12-717a5a239c22
Pollard, Michael
f8b42aac-3373-4a69-9622-cbc5c52f363b
Branlund, Eric
ef835f73-729c-4105-b372-9ec8b50602e1
Sedat, John W
d8c9fd2d-0f08-4e8e-bb16-3fb4a6375434
Agard, David A
c0733eec-3b98-4a05-9cac-4a965769f5b1
Arigovindan, Muthuvel
3b1f076d-191e-4268-9283-689e48b77a33
Fung, Jennifer C.
8617f84c-7e4a-4d1b-898c-b51a600d46d9
Elnatan, Daniel
1065ca8c-5c21-4144-b131-a33417312e02
Mennella, Vito
43c60e29-c0a7-4ab8-8e5c-fcb59f70a28a
Chan, Yee-Hung Mark
9068ef12-a2cf-4d5b-bc12-717a5a239c22
Pollard, Michael
f8b42aac-3373-4a69-9622-cbc5c52f363b
Branlund, Eric
ef835f73-729c-4105-b372-9ec8b50602e1
Sedat, John W
d8c9fd2d-0f08-4e8e-bb16-3fb4a6375434
Agard, David A
c0733eec-3b98-4a05-9cac-4a965769f5b1

Arigovindan, Muthuvel, Fung, Jennifer C., Elnatan, Daniel, Mennella, Vito, Chan, Yee-Hung Mark, Pollard, Michael, Branlund, Eric, Sedat, John W and Agard, David A (2013) High-resolution restoration of 3D structures from widefield images with extreme low signal-to-noise-ratio. Proceedings of the National Academy of Sciences of the United States of America, 110 (43), 17344-17349. (doi:10.1073/pnas.1315675110).

Record type: Article

Abstract

Four-dimensional fluorescence microscopy--which records 3D image information as a function of time--provides an unbiased way of tracking dynamic behavior of subcellular components in living samples and capturing key events in complex macromolecular processes. Unfortunately, the combination of phototoxicity and photobleaching can severely limit the density or duration of sampling, thereby limiting the biological information that can be obtained. Although widefield microscopy provides a very light-efficient way of imaging, obtaining high-quality reconstructions requires deconvolution to remove optical aberrations. Unfortunately, most deconvolution methods perform very poorly at low signal-to-noise ratios, thereby requiring moderate photon doses to obtain acceptable resolution. We present a unique deconvolution method that combines an entropy-based regularization function with kernels that can exploit general spatial characteristics of the fluorescence image to push the required dose to extreme low levels, resulting in an enabling technology for high-resolution in vivo biological imaging.

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More information

e-pub ahead of print date: 8 October 2013
Published date: 22 October 2013
Keywords: Algorithms, Animals, Cell Line, Entropy, Imaging, Three-Dimensional/methods, Microscopy, Fluorescence/methods, Models, Molecular, Models, Theoretical, Nuclear Proteins/chemistry, Protein Conformation, Reproducibility of Results, Saccharomyces cerevisiae/cytology, Saccharomyces cerevisiae Proteins/chemistry, Signal-To-Noise Ratio

Identifiers

Local EPrints ID: 434039
URI: http://eprints.soton.ac.uk/id/eprint/434039
ISSN: 0027-8424
PURE UUID: 1c53a0dc-ed79-456e-8afe-afafd5e4e398
ORCID for Vito Mennella: ORCID iD orcid.org/0000-0002-4842-9012

Catalogue record

Date deposited: 11 Sep 2019 16:30
Last modified: 16 Mar 2024 04:04

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Contributors

Author: Muthuvel Arigovindan
Author: Jennifer C. Fung
Author: Daniel Elnatan
Author: Vito Mennella ORCID iD
Author: Yee-Hung Mark Chan
Author: Michael Pollard
Author: Eric Branlund
Author: John W Sedat
Author: David A Agard

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