Parallel and flexible fluorescent imaging using two-photon RESOLFT super-resolution microscopy with spatial light modulator control

Abstract

High resolution imaging in three dimension is important for biological research. The RESOLFT (Reversible Saturable Optical Fluorescence Transitions) fluorescent microscopy is one technique which can achieve lateral super-resolution imaging. Two-photon microscopy naturally generate high resolution in the longitudinal direction with less background compared to single photon excitation. We combine these two methods to realize three-dimensional high-resolution imaging. This super-resolution method also is limited in imaging speed. We use a spatial light modulator (SLM) as a flexible phase mask of the microscopy. It is used to compensate the system aberration, as well as increasing the imaging speed. The parallel scanning generates multiple super-resolution focuses as an array or in arbitrary positions by phase retrieval calculation. This microscopy combined with SLM control could applied to high throughput 3D imaging or multiple spots tracking in high-resolution.