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Instant square lattice structured illumination microscopy: an optimal strategy towards photon-saving and real-time super-resolution observation
preprintposted on 2024-02-07, 17:00 authored by Tianyu Zhao, Zhaojun Wang, Manming Shu, Jingxiang Zhang, Yansheng Liang, Shaowei Wang, Ming Lei
Over the past decade, structured illumination microscopy (SIM) has found its niche in super-resolution (SR) microscopy due to its fast imaging speed and low excitation intensity. However, due to the significantly higher light dose compared to wide-field microscopy and the time-consuming post-processing procedures, long-term, real-time, super-resolution observation of living cells is still out of reach for most SIM setups, which inevitably limits its routine use by cell biologists. Here, we describe square lattice SIM (SL-SIM) for long-duration live cell imaging by using the square lattice optical field as illumination, which allows continuous super-resolved observation over long periods of time. In addition, by extending the previous joint spatial-frequency reconstruction concept to SL-SIM, a high-speed reconstruction strategy is validated in the GPU environment, whose reconstruction time is even shorter than image acquisition time, thus enabling real-time observation. We have demonstrated the potential of SL-SIM on various biological applications, ranging from microtubule cytoskeleton dynamics to the interactions of mitochondrial cristae and DNAs in COS7 cells. The inherent lower light dose and user-friendly workflow of the SL-SIM could help make long-duration, real-time and super-resolved observations accessible to biological laboratories.