Laser-cytofluorescence microscopic image restoration by iterative deconvolution

Details Laser-cytofluorescence microscopic image restoration by iterative deconvolution Laser-cytofluorescence microscopic image restoration by iterative deconvolution

Jan 1995

adsabs.harvard.edu/cgi-bin/nph-data_query?bibcode=1995spie.2324..215x&link_type=abstract

Proc. SPIE Vol. 2324, p. 215-222, Optical Biopsy and Fluorescence Spectroscopy and Imaging, Rinaldo Cubeddu; Renato Marchesini;

Computer Science

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Scientific paper

The aim of our study is to improve the performances of an optical microscope by image deconvolution technique to attain that of a confocal one in the field of laser cytofluorescence. The fluorescence of antigen lymphocyte sites marked by rhodamine is induced by a laser ((lambda) equals 543 nm) or a mercury vapor lamp. A set of scanned fluorescence images is acquired at different focal planes by a SIT camera, fitted on an optical ZEISS microscope (N.A. 1.25, X100). The entire system is controlled by a PC equipped with a MATROX-MVP-AT image processing card. Since an optical microscope has a more important focal depth than a confocal one, an improved iterative deconvolution algorithm of Van Cittert's has been used to artificially reduce the focal depth. In order to ensure the convergence of such an iterative algorithm, a new criterion for relaxation coefficient is defined through a theoretical study. The experiments show an improvement of 50% in the spatial x-y resolution of 30% in optical z axis gain. As regards our application, this processing brings our system to a comparable performance level as a confocal microscope.

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