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          Institute: MPI für biophysikalische Chemie     Collection: Molekulare Biologie (Dr. Thomas M. Jovin)     Display Documents

ID: 42767.0, MPI für biophysikalische Chemie / Molekulare Biologie (Dr. Thomas M. Jovin)
Resolution enhancement by subtraction of confocal signals taken at different pinhole sizes
Authors:Heintzmann, R.; Sarafis, V.; Munroe, P.; Nailon, J.; Hanley, Q. S.; Jovin, T. M.
Date of Publication (YYYY-MM-DD):2003
Title of Journal:Micron
Start Page:293
End Page:300
Copyright:c*2003 Elsevier Ltd.
Review Status:Peer-review
Audience:Not Specified
Abstract / Description:Subtractive imaging in confocal fluorescence light microscopy is based on the subtraction of a suitably scaled widefield image from a confocal image. An approximation to a widefield image can be obtained by detection with an opened confocal pinhole. The subtraction of images enhances the resolution in-plane as well as along the optic axis. Due to the linearity of the approach, the effect of subtractive imaging in Fourier-space corresponds to a suppression of low spatial frequencies leading to a relative enhancement of the high frequencies. Along the direction of the optic axis this also results in an improved sectioning. Image processing can achieve a similar effect. However, a 3D volume dataset must be acquired and processed, yielding a result essentially identical to subtractive imaging but superior in signal to noise ratio (SNR). The latter can be increased further with the technique of weighted averaging in Fourier-space. A comparison of 2D and 3D experimental data analysed with subtractive imaging, the equivalent Fourier-space processing of the confocal data only, and Fourier-space weighted averaging is presented.
Free Keywords:Subtractive Imaging, Superresolution, Composite Image, Fourier-space Weigthing, Weighted Averaging
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für biophysikalische Chemie/Abt. Thomas Jovin / 060
External Affiliations:School of Computing and Electrical Engineering, University of Queensland, Qld 4072, Australia; Electron Microscopy Unit, University of New South Wales, Sydney, NSW 2052, Australia; Centre for Microscopy and Microanalysis, University of Queensland, Qld 4072, Australia; Department of Biological Sciences, University of the West Indies, Cave Hill Campus, St. Michael, Barbados
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