Numerical analysis of resolution enhancement in laser scanning microscopy using a radially polarized beam

The spatial resolution characteristics in confocal laser scanning microscopy (LSM) and two-photon LSM utilizing a higher-order radially polarized Laguerre-Gaussian (RP-LG) beam are numerically analyzed. The size of the point spread function (PSF) and its dependence on the confocal pinhole size are compared with practical LSM using a circularly polarized Gaussian beam on the basis of vector diffraction theory. The spatial frequency response in terms of the optical transfer function (OTF) is also evaluated for LSM using the RP-LG beam. The smaller focal spot characteristics of higher-order RP-LG beams contribute to a dramatic enhancement of the lateral spatial resolution in confocal LSM and two-photon LSM.