Regular papers

Multi-kernel deconvolution applied to confocal fluorescence microscopy with engineered point spread function

Laboratoire MIPS–Groupe Lab.El, Université de Haute–Alsace, IUT Mulhouse, 61 rue A. Camus F– 68093 Mulhouse Cedex France

^* bertrand.simon@uha.fr
^** olivier.haeberle@uha.fr

Received: 16 October 2006

Abstract

Fluorescence microscopy is a powerful technique in biology, because of the immense variety of markers now available. Compared to other methods, its resolution is however limited. In wide–field microscopy, the technique of structured illumination permits to improve the lateral resolution by a factor of two, even surpassing confocal microscopy, which permits a theoretical gain of about 40%. We propose an alternate technique, combining laterally interfering focused beams, which should permit the same gain of resolution in a confocal microscope. Furthermore, this technique, combined with multiple acquisition and multikernel deconvolution, permits a better object reconstruction than classical monokernel deconvolution using a regular excitation point spread function.