In point-scanning microscopy, such as confocal laser scanning microscopy (CLSM), the imaging resolution is limited by the size of the focal spot. Breaking through the diffraction limit by inserting a modulation phase mask in the objective pupil plane is a compact, high efficiency and adaptable scheme. Here, we propose a feedback phase design method based on genetic algorithm to control the 3D shape of the focal spot. The lateral and axial size can be jointly designed for various usages. Simulation and experimental results present the feasibility of this method. The axial size can be decreased by 50% while the lateral size is decreased by 15% simultaneously. This method of 3D super-diffraction limit focusing may potentially be used in CLSM for 3D super resolution imaging.
Optical microscopy is an indispensable imaging tool in life science. However, light cannot be focused owing to high scattering, and hence, the imaging depth and spatial resolution are restricted. Here, we propose an imaging method that combines wavefront shaping and image scanning microscopy. The reflected signal is used as feedback to acquire an optimal phase that can refocus the scattered light behind the scattering media. The experimental results show that the proposed method works in multilayer scattering media and can improve both the resolution and imaging depth of optical microscopy.
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