Three-Dimensional Image Restoration In Fluorescence Confocal Scanning Microscopy

M. Bertero; P. Boccacci; G. J. Brakenhoff; F. Malfanti; H. T. M. van der Voort

doi:10.1117/12.961778

28 September 1989 Three-Dimensional Image Restoration In Fluorescence Confocal Scanning Microscopy

M. Bertero, P. Boccacci, G. J. Brakenhoff, F. Malfanti, H. T. M. van der Voort

Proceedings Volume 1139, Optical Storage and Scanning Technology; (1989) https://doi.org/10.1117/12.961778
Event: 1989 International Congress on Optical Science and Engineering, 1989, Paris, France

Abstract

In the case of weakly absorbing objects and of complete incoherence of the emitted radiation, the three-dimensional image g(r) produced by a Confocal Scanning Laser Microscope (CSLM) is the convolution of the distribution function of the fluorescent material, f(r), with the impulse response function of the instrument, h(r). Since h(r) is band-limited, the image g(r) contains information only about the Fourier components of f(r) in the band of h(r). Moreover, these components are distorted because the Fourier transform of h(r) is not constant over the band. If both g(r) and h(r) are known it is possible to invert the integral relationship indicated above in order to improve the imaging fidelity and the resolution of the microscope. This deconvolution problem is ill-posed and the so-called regularization methods must be used in order to get stable and approximate solutions. In this paper we discuss the potential applications of a very simple regularization technique to the restoration of three-dimensional images produced by the confocal scanning microscope developed at the University of Amsterdam.

Citation Download Citation

M. Bertero, P. Boccacci, G. J. Brakenhoff, F. Malfanti, and H. T. M. van der Voort "Three-Dimensional Image Restoration In Fluorescence Confocal Scanning Microscopy", Proc. SPIE 1139, Optical Storage and Scanning Technology, (28 September 1989); https://doi.org/10.1117/12.961778

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