Utilizing phase-modulated Lissajous to enhance spatial and temporal resolution of laser scanning microscopy

Abstract

Abstract One of the main challenges in scanning microscopy is increasing the scanning speed without deteriorating the spatial resolution of the image. With Lissajous scanning, the image is available in a fraction of the time since it gathers scattered data from the entire field of view. Nevertheless, this method reduces the density of scan lines at the center of the image, thereby reducing the imaging resolution. This problem can be solved by phase modulation of the motion function of the scanner; however, the density of lines created by such designs is not always controllable. In this paper, simple relationships for determining the desired density over a large area are presented for selecting the appropriate phase modulation, in particular for microscopy applications. In addition, a confocal microscope was employed to image samples using the proposed relations to obtain temporal and spatial resolution two times higher than conventional Lissajous. The results demonstrated that the imaging speed can be significantly increased by using phase modulation accompanied by interpolation methods for image reconstruction.