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volumetric imaging of Neural Activity by Light Field Microscopy

Neuroscience Bulletin 2022年第12期38卷 1559-1568页

作者： Lu Bai Zhenkun Zhang Lichen Ye Lin Cong Yuchen Zhao Tianlei Zhang Ziqi Shi Kai Wang Institute of Neuroscience State Key Laboratory of NeuroscienceCenter for Excellence in Brain Science and Intelligence TechnologyChinese Academy of SciencesShanghai 200031China University of Chinese Academy of Sciences Beijing 100049China Shanghai Center for Brain Science and Brain-Inspired Intelligence Technology Shanghai 201210China

Recording the highly diverse and dynamic activities in large populations of neurons in behaving animals is crucial for a better understanding of how the brain works.To meet this challenge,extensive efforts have been devoted to developing functional fluorescent indicators and optical imaging techniques to optically monitor neural activity.Indeed,optical imaging potentially has extremely high throughput due to its non-invasive access to large brain regions and capability to sample neurons at high density,but the readout speed,such as the scanning speed in two-photon scanning microscopy,is often limited by various practical considerations.Among different imaging methods,light field microscopy features a highly parallelized 3D fluorescence imaging scheme and therefore promises a novel and faster strategy for functional imaging of neural activity.Here,we briefly review the working principles of various types of light field microscopes and their recent developments and applications in neuroscience studies.We also discuss strategies and considerations of optimizing light field microscopy for different experimental purposes,with illustrative examples in imaging zebrafish and mouse brains.

关键词： Light feld microscopy volumetric imaging Brain activity Calcium imaging Voltage imaging

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Single-cell volumetric imaging with light field microscopy: Advances in systems and algorithms

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Journal of Innovative Optical Health Sciences 2023年第2期16卷 58-74页

作者： Beibei Gao Lu Gao Fu Wang School of Biomedical Engineering Shanghai Jiao Tong University Shanghai 200240 P.R.China

Single-cell volumetric imaging is essential for researching individual characteristics of cells.As a nonscanning imaging technique,lighteld microscopy(LFM)is a critical tool to achieve realtime three-dimensional imaging with the advantage of single-shot.To address the inherent limits including nonuniform resolution and block-wise artifacts,various modied LFM strategies have been developed to provide new insights into the structural and functional information of cells.This review will introduce the principle and development of LFM,discuss the improved approaches based on hardware designs and 3D reconstruction algorithms,and present the applications in single-cell imaging.

关键词： Light field microscopy single-cell imaging volumetric imaging 3D reconstruction

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In vivo volumetric imaging of fast biological dynamics in deep tissue via wavefront engineering

In vivo volumetric imaging of fast biological dynamics in de...

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中国化学会第十三届全国分析化学年会

作者： Lingjie Kong Department of Precision Instruments Tsinghua University

High-speed volumetric imaging of biological dynamics in deep tissue in vivo is highly desired in biomedical studies, such as cellular interactions and signaling dynamics across large cell populations. Here we exploit wavefront engineering as the solution of volumetric imaging of fast biological dynamics in deep tissue in vivo. To compensate the wavefront distortion induced by the aberration and random scattering in heterogeneous tissues, we have developed the Iterative MultiPhoton Adaptive Compensation Technique for wavefront measurement and compensation, and demonstrated functional imaging of dendritic spines through intact mouse skull at high resolution, for the first time. Recently, we developed Multi-Pupil Adaptive Optics for simultaneously compensating wavefront distortions over a large field-of-view(FOV). With wavefront compensation over 9 pupils in parallel, we demonstrated high-speed functional imaging of neural network over a 450 × 450 μm FOV at large depth in mouse cerebral cortex. On high-speed volumetric imaging, we developed an optical phase-locked ultrasound lens to generate oscillating defocusing wavefront at ～450 kHz and achieved MHz axial scanning, several orders faster than that of other techniques, which enabled volumetric imaging at volume rate up to hundreds of Hz. We also integrated the wavefront engineering techniques proposed above, and demonstrated functional imaging of neuron network in awake, behaving animals and dynamical imaging of immune cells trafficking in the circulation system in vivo.

关键词： multiphoton microscopy wavefront engineering adaptive optics volumetric imaging three-dimensional microscopy functional imaging

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A simple Bessel module with tunable focal depth and constant resolution for commercial two-photon microscope

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创新光学健康科学杂志(英文) 2023年第4期16卷 150-159页

作者： Ting Mo Yiran Liu Fatao Bie Zimin Dai Jin Chang Hui Gong Wei Zhou Britton Chance Center and MoE Key Laboratory for Biomedical Photonics Wuhan National Laboratory for Optoelectronics Huazhong University of Science and Technology Wuhan P.R.China MoE Key Laboratory for Biomedical Photonics Collaborative Innovation Center for Biomedical Engineering School of Engineering Sciences Huazhong University of Science and Technology Wuhan Hubei 430074P.R.China Research Unit of Multimodal Cross Scale Neural Signal Detection and imaging Chinese Academy of Medical Sciences HUST-Suzhou Institute for Brainsmatics JITRI Suzhou P.R.China

The volumetric imaging of two-photon microscopy expands the focal depth and improves the throughput,which has unparalleled superiority for three-dimension samples,especially in neuroscience.However,emerging in volumetric imaging is still largely customized,which limits the integration with commercial two-photon systems.Here,we analyzed the key parameters that modulate the focal depth and lateral resolution of polarized annular imaging and proposed a volumetric imaging module that can be directly integrated into commercial two-photon systems using conventional optical elements.This design incorporates the beam diameter adjustment settings of commercial two-photon systems,allowing flexibility to adjust the depth of focus while maintaining the same lateral resolution.Further,the depth range and lateral resolution of the design were verified,and the imaging throughput was demonstrated by an increase in the number of imaging neurons in the awake mouse cerebral cortex.

关键词： volumetric imaging Bessel beam two-photon microscopy

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Multiscale label-free volumetric holographic histopathology of thick-tissue slides with subcellular resolution

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Advanced Photonics 2021年第2期3卷 98-105页

作者： Herve Hugonnet Yeon Wook Kim Moosung Lee Seungwoo Shin Ralph H.Hruban Seung-Mo Hong YongKeun Park Korea Advanced Institute of Science and Technology Department of PhysicsDaejeonRepublic of Korea KAIST Institute for Health Science and Technology KAISTDaejeonRepublic of Korea Asan Institute for Life Science Asan Medical CenterSeoulRepublic of Korea Johns Hopkins Medical Institutions Sol Goldman Pancreatic Cancer Research CenterDepartment of PathologyBaltimoreMarylandUnited States University of Ulsan College of Medicine Asan Medical CenterDepartment of PathologySeoulRepublic of Korea Tomocube Inc. DaejeonRepublic of Korea

Histopathology relies upon the staining and sectioning of biological tissues,which can be laborious and may cause artifacts and distort tissues.We develop label-free volumetric imaging of thick-tissue slides,exploiting refractive index distributions as intrinsic imaging contrast.The present method systematically exploits label-free quantitative phase imaging techniques,volumetric reconstruction of intrinsic refractive index distributions in tissues,and numerical algorithms for the seamless stitching of multiple three-dimensional tomograms and for reducing scattering-induced image distortion.We demonstrated label-free volumetric imaging of thick tissues with the field of view of 2 mm×1.75 mm×0.2 mm with a spatial resolution of 170 nm×170 nm×1400 nm.The number of optical modes,calculated as the reconstructed volume divided by the size of the point spread function,was∼20 giga voxels.We have also demonstrated that different tumor types and a variety of precursor lesions and pathologies can be visualized with the present method.

关键词： optical diffraction tomography histology volumetric imaging label-free imaging wide field of view

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