Cross structured illumination for high speed high resolution line scanning confocal microscopy

Abstract

Previous research presented the structured illumination confocal scanning microscope (SICSM) so as to improve the lateral resolution of the confocal microscope. However, the image acquisition speed of the SICSM is very slow and also an alignment error due to the mechanical rotation of a grating and a slit can easily occur. As a theoretical study, in this paper we propose a new SI method, the cross SI method, which improves lateral resolution and image acquisition speed. Performances of the conventional SI and the proposed SI methods are compared by analysis of the modulation transfer function. The proposed SI method shows similar lateral resolution and can shorten the image acquisition time compared to the conventional SI method. The cross structured illumination confocal microscope (CSICM) is combined with the cross SI pattern optics and the line scanning confocal microscope. We have introduced a 2-D diffractive grating, four linear polarizers and four cylindrical lenses in order to create the cross SI pattern. The effects of the cross SI pattern, intensity and visibility, on the system performance are analyzed. The CSICM has double the lateral resolution of the conventional microscope, an optical sectioning ability and a fast image acquisition speed.