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ID: 17040.0, MPI für biophysikalische Chemie / Molekulare Biologie (Dr. Thomas M. Jovin)
Axial tomographic confocal fluorescence microscopy
Authors:Heintzmann, R.; Cremer, C.
Language:English
Date of Publication (YYYY-MM-DD):2002-04
Title of Journal:Journal of Microscopy-Oxford
Volume:206
Start Page:7
End Page:23
Review Status:Peer-review
Audience:Not Specified
Abstract / Description: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.
Free Keywords:acceleration; axial resolution; CLSM; combined ML- deconvolution; confocal
Comment of the Author/Creator:Date: 2002, APR
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für biophysikalische Chemie/Abt. Thomas Jovin / 060
External Affiliations:Univ Heidelberg, Kirchhoff Inst Phys, D-69120 Heidelberg, Germany
Identifiers:URL:http://onlinelibrary.wiley.com/doi/10.1046/j.1365-...
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