Fast single-layer reconstruction for three-dimensional structured illumination microscopy

Abstract

Although three-dimensional structured illumination microscopy (3D-SIM) that enhances the spatial resolution of widefield microscope in three dimensions has been successfully commercialized as a variety of products, the low imaging speed impedes its application in fast subcellular dynamics. To adapt to fast dynamics, most 3D-SIM setups provide an option for the users by canceling the axial scanning during the acquisition, by which two-dimensional optically-sectioned, super-resolved (OS-SR) time series can be obtained with single-layer 3D-SIM data. However, the conventional reconstruction workflow implemented in the frequency domain generally takes seconds for every single reconstruction, which nullifies real-time feedback for these 3D-SIM setups. In this paper, we establish a fast single-layer reconstruction approach for 3D-SIM by extending a high-speed reconstruction framework, joint space-frequency reconstruction (JSFR), to single-layer 3D-SIM. Consequently, the reconstruction speed is improved over the conventional method, without compromising the image quality. This approach is expected to provide a useful tool for 3D-SIM microscope manufacturers and their users to achieve real-time, single-layer imaging of living cells.