Resolution enhancement by subtraction of confocal signals taken at different pinhole sizes

Heintzmann, Rainer, Sarafis, Vassilios, Munroe, Paul, Nailon, John, Hanley, Quentin S. and Jovin, Thomas M. (2003) Resolution enhancement by subtraction of confocal signals taken at different pinhole sizes. Micron, 34 6-7: 293-300. doi:10.1016/S0968-4328(03)00054-4


Author Heintzmann, Rainer
Sarafis, Vassilios
Munroe, Paul
Nailon, John
Hanley, Quentin S.
Jovin, Thomas M.
Title Resolution enhancement by subtraction of confocal signals taken at different pinhole sizes
Journal name Micron   Check publisher's open access policy
ISSN 0968-4328
Publication date 2003-10
Sub-type Critical review of research, literature review, critical commentary
DOI 10.1016/S0968-4328(03)00054-4
Volume 34
Issue 6-7
Start page 293
End page 300
Total pages 8
Place of publication Oxford, England
Publisher Pergamon-Elsevier Science Ltd.
Collection year 2003
Language eng
Subject C1
230108 Harmonic and Fourier Analysis
671401 Scientific instrumentation
Abstract Subtractive imaging in confocal fluorescence light microscopy is based on the subtraction of a suitably weighted 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 reduction of low spatial frequency contributions 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. 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. (C) 2003 Elsevier Ltd. All rights reserved.
Keyword Microscopy
Subtractive Imaging
Superresolution
Composite Image
Fourier-space Weighting
Weighted Averaging
Array Microscope Pam
Fluorescence Microscopy
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Document type: Journal Article
Sub-type: Critical review of research, literature review, critical commentary
Collections: 2004 Higher Education Research Data Collection
School of Chemistry and Molecular Biosciences
 
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Created: Wed, 15 Aug 2007, 01:51:57 EST