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  Axial tomographic confocal fluorescence microscopy

Heintzmann, R., & Cremer, C. (2002). Axial tomographic confocal fluorescence microscopy. Journal of Microscopy-Oxford, 206, 7-23. Retrieved from http://onlinelibrary.wiley.com/doi/10.1046/j.1365-2818.2002.01000.x/pdf.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0012-F408-7 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0027-E581-B
Genre: Journal Article

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 Creators:
Heintzmann, R.1, Author              
Cremer, C., Author
Affiliations:
1Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society, ou_578628              

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Free keywords: acceleration; axial resolution; CLSM; combined ML- deconvolution; confocal
 Abstract: By physical rotation of the sample, axial tomography enables the acquisition of otherwise inaccessible spatial information from an object. In combination with confocal microscopy, the method can fundamentally improve the effective three- dimensional (3D) resolution. In this report we present a novel method for high resolution reconstruction of confocal axial tomographic data. The method automatically determines the relative angles of rotation, aligns the data from different rotational views and reconstructs a single high resolution 3D dataset. The reconstruction makes use of a known point spread function and is based on an unconstrained maximum likelihood deconvolution performed simultaneously from multiple (in our case three) angular views. It was applied to simulated as well as to experimental confocal datasets. The gain in resolution was quantified and the effect of choice of overrelaxation factors on the speed of convergence was investigated. A clearly improved 3D resolution was obtained by axial tomography together with reconstruction as compared with reconstruction of confocal data from only it single angular view.

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Language(s): eng - English
 Dates: 2002-04
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Degree: -

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Title: Journal of Microscopy-Oxford
Source Genre: Journal
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Publ. Info: -
Pages: - Volume / Issue: 206 Sequence Number: - Start / End Page: 7 - 23 Identifier: -