Structured illumination microscopy(SIM)is an established optical superresolution imaging technique.However,conventional SIM based on wide-field image acquisition is generally limited to visualizing thin cellular sampl...Structured illumination microscopy(SIM)is an established optical superresolution imaging technique.However,conventional SIM based on wide-field image acquisition is generally limited to visualizing thin cellular samples.We propose combining one-dimensional image rescan and structured illumination in the orthogonal direction to achieve superresolution without the need to rotate the illumination pattern.The image acquisition speed is consequently improved threefold,which is also beneficial for minimizing photobleaching and phototoxicity.Optical sectioning in thick biological tissue is enhanced by including a confocal slit in the system to significantly suppress the out-of-focus background and the associated noise.With all the technical improvements,our method captures threedimensional superresolved image stacks of neuronal structures in mouse brain tissue samples for a depth range of more than 200μm.展开更多
基金supported by the Ministry of Education-Singapore(Grant Nos.MOE2019-T2-2-094 and MOE Tier I R-397-000-327-114)Shenzhen Science and Technology Program(Grant No.GJHZ20210705141805015).
文摘Structured illumination microscopy(SIM)is an established optical superresolution imaging technique.However,conventional SIM based on wide-field image acquisition is generally limited to visualizing thin cellular samples.We propose combining one-dimensional image rescan and structured illumination in the orthogonal direction to achieve superresolution without the need to rotate the illumination pattern.The image acquisition speed is consequently improved threefold,which is also beneficial for minimizing photobleaching and phototoxicity.Optical sectioning in thick biological tissue is enhanced by including a confocal slit in the system to significantly suppress the out-of-focus background and the associated noise.With all the technical improvements,our method captures threedimensional superresolved image stacks of neuronal structures in mouse brain tissue samples for a depth range of more than 200μm.
