The slow axial scanning rate in multiphoton microscopy(MPM)has traditionally limited the speed of three-dimensional(3D)imaging.Recently,a lot of techniques have been proposed to speed up the axial scan;however,there i...The slow axial scanning rate in multiphoton microscopy(MPM)has traditionally limited the speed of three-dimensional(3D)imaging.Recently,a lot of techniques have been proposed to speed up the axial scan;however,there inherently exists an upper limit of the achievable maximum scanning rate restricted by full sampling.To overcome this limitation,we developed an approach to realize multiplane compressive imaging in MPM that empowers conventional laser scanning microscopies with rapid axial scanning capacity in a sub-sampling way.To realize the technique,we achieved two technical breakthroughs:first,we proposed a concept to axially encode the beam with binary intensities;second,compressive sensing theory was introduced to the axial direction in MPM based on the axial-coded point spread function.This 3D imaging technology is termed arbitrary illumination microscopy with encoded depth(AIMED),enabling a nearly double volumetric imaging speed with subcellular resolution for mouse brain neurons in experiments and performing approximately eight times faster in simulation.The axial compressive ability of AIMED can be readily extended to other microscopic modalities for achieving axially compressive 3D imaging.Our concepts demonstrated provide insights into the entire field of advanced volumetric microscopy.展开更多
基金supported by the Research Grants Council of the Hong Kong Special Administrative Region of China(Grant Nos.HKU 17210522,HKU C7074-21G,HKU R7003-21,HKU 17205321,and HKU 17200219)the Innovation and Technology Commission of the Hong Kong SAR Government(Grant Nos.MHP/073/20,MHP/057/21,and Health@Inno HK program)+2 种基金the National Natural Science Foundation of China(Grant No.62305274)the Natural Science Foundation of Xiamen City,China(Grant No.3502Z202371001)the Fujian Provincial Natural Science Foundation(Grant No.2024J01056)。
文摘The slow axial scanning rate in multiphoton microscopy(MPM)has traditionally limited the speed of three-dimensional(3D)imaging.Recently,a lot of techniques have been proposed to speed up the axial scan;however,there inherently exists an upper limit of the achievable maximum scanning rate restricted by full sampling.To overcome this limitation,we developed an approach to realize multiplane compressive imaging in MPM that empowers conventional laser scanning microscopies with rapid axial scanning capacity in a sub-sampling way.To realize the technique,we achieved two technical breakthroughs:first,we proposed a concept to axially encode the beam with binary intensities;second,compressive sensing theory was introduced to the axial direction in MPM based on the axial-coded point spread function.This 3D imaging technology is termed arbitrary illumination microscopy with encoded depth(AIMED),enabling a nearly double volumetric imaging speed with subcellular resolution for mouse brain neurons in experiments and performing approximately eight times faster in simulation.The axial compressive ability of AIMED can be readily extended to other microscopic modalities for achieving axially compressive 3D imaging.Our concepts demonstrated provide insights into the entire field of advanced volumetric microscopy.
