Multi-color super-resolution microscopy with AOTF-assisted super-resolution optical fluctuation imaging

Multi-color resolution is an indispensable method in biological experiments. However, in the actual experiment, due to the wide fluorescence spectrum and the proximity of multiple fluorescence wavelengths, the crosstalk rate is high. As a result, researchers often cannot distinguish multiple different markers with the naked eye. In order to solve the problem of indistinguishable multi-target markers and limited resolution, we propose a multi-color super-resolution imaging method based on AOTF-assisted super-resolution optical fluctuation imaging. The emission intensity of each fluorophore is different under different excitation light. It has the characteristic of specific excitation spectrum. By using AOTF to scan a wide range of excitation wavelengths, different fluorophore groups are linearly superposed in the wide-field image. In this paper, the abundance distribution of each fluorophore is calculated by combining the separation algorithm for solving the linear equation with multicolor fluorescence microscope. The abundance distribution is used as the image of each color channel to achieve multicolor resolution. In addition, super resolution imaging can be achieved by combining with fluorescence microscope. Simple numerical simulation results show that the image resolution is improved by root 2 times and low spectral crosstalk rate is achieved by combining the two methods.