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

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.