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Subvoxel light-sheet microscopy for high-resolution high-throughput volumetric imaging of large biomedical specimens

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Advanced Photonics 2019年第1期1卷 45-57页

作者： Peng Fei Jun Nie Juhyun Lee Yichen Ding Shuoran Li Hao Zhang Masaya Hagiwara Tingting Yu Tatiana Segura Chih-Ming Ho Dan Zhu Tzung KHsiaic Huazhong University of Science and Technology School of Optical and Electronic InformationWuhanChina Huazhong University of Science and Technology Britton Chance Center for Biomedical PhotonicsWuhan National Laboratory for OptoelectronicsWuhanChina University of California Los AngelesDepartment of BioengineeringLos AngelesCaliforniaUnited States University of Texas at Arlington Joint Department of Bioengineering of UT Arlington/UT SouthwesternArlingtonTexasUnited States University of California Los AngelesSchool of MedicineLos AngelesCaliforniaUnited States University of California Los AngelesChemical and Biomolecular Engineering DepartmentLos AngelesCaliforniaUnited States Osaka Prefecture University Nanoscience and Nanotechnology Research CenterResearch Organization for the 21st CenturyOsakaJapan University of California Los AngelesMechanical and Aerospace Engineering DepartmentLos AngelesCaliforniaUnited States

A key challenge when imaging whole biomedical specimens is how to quickly obtain massive cellular information over a large field of view(FOV).We report a subvoxel light-sheet microscopy(SLSM)method enabling high-throughput volumetric imaging of mesoscale specimens at cellular resolution.A nonaxial,continuous scanning strategy is developed to rapidly acquire a stack of large-FOV images with three-dimensional(3-D)nanoscale shifts encoded.Then,by adopting a subvoxel-resolving procedure,the SLSM method models these low-resolution,cross-correlated images in the spatial domain and can iteratively recover a 3-D image with improved resolution throughout the sample.This technique can surpass the optical limit of a conventional light-sheet microscope by more than three times,with high acquisition speeds of gigavoxels per minute.By fast reconstruction of 3-D cultured cells,intact organs,and live embryos,SLSM method presents a convenient way to circumvent the trade-off between mapping large-scale tissue(>100 mm3)and observing single cell(∼1-μm resolution).It also eliminates the need of complicated mechanical stitching or modulated illumination,using a simple light-sheet setup and fast graphics processing unit-based computation to achieve high-throughput,high-resolution 3-D microscopy,which could be tailored for a wide range of biomedical applications in pathology,histology,neuroscience,etc.

关键词： light-sheet microscopy subvoxel-resolving reconstruction large tissue imaging high-throughput volumetric imaging

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Effects of delayed repair of peripheral nerve injury on the spatial distribution of motor endplates in target muscle

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Neural Regeneration Research 2022年第2期17卷 459-464页

作者： Dong-Dong Li Jin Deng Bo Jin Shuai Han Xin-Yi Gu Xue-Feng Zhou Xiao-Feng Yin Department of Trauma and Orthopedics Peking University People’s HospitalBeijingChina Department of Orthopedics PLA Strategic Support Force Medical CenterBeijingChina

Motor endplates(MEPs) are important sites of information exchange between motor neurons and skeletal muscle, and are distributed in an organized pattern of lamellae in the muscle. Delayed repair of peripheral nerve injury typically results in unsatisfactory functional recovery because of MEP degeneration. In this study, the mouse tibial nerve was transected and repaired with a biodegradable chitin conduit, immediately following or 1 or 3 months after the injury. Fluorescent α-bungarotoxin was injected to label MEPs. Tissue optical clearing combined with light-sheet microscopy revealed that MEPs were distributed in an organized pattern of lamellae in skeletal muscle after delayed repair for 1 and 3 months. However, the total number of MEPs, the number of MEPs per lamellar cluster, and the maturation of single MEPs in gastrocnemius muscle gradually decreased with increasing denervation time. These findings suggest that delayed repair can restore the spatial distribution of MEPs, but it has an adverse effect on the homogeneity of MEPs in the lamellar clusters and the total number of MEPs in the target muscle. The study procedures were approved by the Animal Ethics Committee of the Peking University People's Hospital(approval No. 2019 PHC015) on April 8, 2019.

关键词： degeneration delayed repair lamellar cluster light-sheet microscopy motor endplates peripheral nerve injury three-dimensional distribution tissue optical clearing

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What we know about axons in Parkinson’s disease

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Aging Communications 2023年第1期5卷 20-28页

作者： Weili Cai Tingyan Wu Weidong Le Xi Chen School of Medical Technology Jiangsu College of NursingHuai'an 22305China Institute of Neurology Sichuan Provincial People’s HospitalUniversity of Electronic Science and Technology of ChinaChengdu 610072China Chinese Academy of Sciences Sichuan Translational Medicine Research Hospital Chengdu 610072China.

Tremendous research efforts have been made regarding the pathogenesis of Parkinson’s disease(PD).However,there are still no effective strategies to restore midbrain dopaminergic(mDA)innervation and prevent disease progression.One possibility is that we may have been neglecting the role of axons in mDA neuronal degeneration.This review first summarizes mDA axon development during the early stage of PD and discusses how axon guidance defects contribute to PD vulnerability.Furthermore,we review axonal transport dysregulation in the numerous PD-related genetic mutations,including Parkin,PINK1,DJ1,LRRK2 and SNCA.The evidence suggests that proper axonal transport is crucial for neuronal function and survival.Finally,advanced tools for axonal studies were evaluated,including light-sheet and super-resolution microscopy.These adapted microscopes have been used to help solve questions unanswered before.Overall,the role of axon terminals in the initiation of the degeneration cascade remains undeciphered,and more research in the related area may be conducted further to restore dopamine levels in the striatum to alleviate the motor complications of PD.

关键词： axon guidance axonal transport Parkinson’s disease light-sheet microscopy super-resolution microscopy

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