Enhanced 3-D reconstruction from confocal scanning microscope images 1: Deterministic and maximum likelihood reconstructions

Abstract

Significant improvement in the longitudinal (optical axis) resolution of a microscope has previously been obtained either by posterior digital processing of optical sections collected with a conventional microscope, or by collecting sections with a confocal scanning microscope. In this paper we report the feasibility of obtaining longitudinal resolution comparable to the lateral diffraction limit by posterior processing of confocal sections. A confocal through-focus image series was simulated numerically and restored by constrained iterative deconvolution and by maximum likelihood. Several typical imaging situations were simulated. The results support the possibility of achieving equal longitudinal and lateral resolution.