Parallel illumination for depletion microscopy through acousto-optic spatial light modulation

Abstract

State-of-the-art super-resolution microscopy techniques, including Stimulated Emission Depletion (STED), Reversible Saturable Optical Fluorescence Transitions (RESOLFT), and Switching Laser Mode (SLAM) microscopies, implement Laguerre-Gaussian beams, also known as vortex or doughnut beams to capture fluorescence information within a sub-wavelength area of the observed sample, effectively surpassing the diffraction limit and significantly improving the quality of the image. However, these techniques typically operate at point by point basis, involving time-consuming scanning of the sample to construct a complete, meaningful image. Therefore, for real-time live cell imaging purposes, the parallelization of illumination is crucial. In this study, we demonstrate the parallel generation of arbitrary arrays of Gaussian and Laguerre-Gaussian laser foci suitable for super-resolution microscopy. We achieve rapid scanning through the sample using acousto-optic spatial light modulation, a technique we have previously pioneered across various fields. By employing parallelized illumination with both Gaussian and doughnut beams, we aim to capture super-resolution images.