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Optical scattering poses a significant challenge to high-resolution microscopy within deep tissue. To accurately predict the performance of various microscopy techniques in thick samples, we present a computational model that efficiently solves Maxwell’s equation in highly scattering media. This toolkit simulates the deterioration of the laser beam point spread function (PSF) without making a paraxial approximation, enabling accurate modeling of high-numerical-aperture (NA) objective lenses commonly employed in experiments. Moreover, this framework is applicable to a broad range of scanning microscopy techniques including confocal microscopy, stimulated emission depletion (STED) microscopy, and ground-state depletion microscopy. Notably, the proposed method requires only readily obtainable macroscopic tissue parameters. As a practical demonstration, we investigate the performance of Laguerre–Gaussian (LG) versus Hermite–Gaussian (HG) depletion beams in STED microscopy. 

We numerically compare the null quality for STED microscopy generated by Laguerre-Gaussian beams with orbital angular momentum and donut beams generated by incoherent addition of orthogonal Hermite Gaussian beams when imaging deep biological tissue.