Scalable volumetric imaging for ultrahigh-speed brain mapping at synaptic resolution  

作　　者：Hao Wang Qingyuan Zhu Lufeng Ding Yan Shen Chao-Yu Yang Fang Xu Chang Shu Yujie Guo Zhiwei Xiong Qinghong Shan Fan Jia Peng Su Qian-Ru Yang Bing Li Yuxiao Cheng Xiaobin He Xi Chen Feng Wu Jiang-Ning Zhou Fuqiang Xu Hua Han Pak-Ming Lau Guo-Qiang Bi 

机构地区：[1]Hefei National Laboratory for Physical Sciences at the Microscale,and School of Life Sciences,University of Science and Technology of China,Hefei 230027,China [2]CAS Key Laboratory of Brain Function and Disease,and School of Life Sciences,University of Science and Technology of China.Hefei 230027,China [3]Institute of Automation,Chinese Academy of Sciences,Beijing 100190,China [4]University of Chinese Academy of Sciences,Beijing 100049,China [5]School of Information Science and Technology,University of Science and Technology of China,Hefei 230027,China [6]National Engineering Laboratory for Brain-inspired Intelligence Technology and Application.University of Science and Technology of China,Hefei 230027,China [7]Wuhan Institute of Physics and Mathematics,Chinese Academy of Sciences,Wuhan 430071,China [8]CAS Center for Excellence in Brain Science and Intelligence Technology,Shanghai 200031,China

出　　处：《National Science Review》2019年第5期982-992,共11页国家科学评论（英文版）

基　　金：supported by the Strategic Priority Research Program of the Chinese Academy of Sciences(XDB02050000to G.-Q.B.,XDB02060000 to F.X.,XDB02030000 to J.-N.Z.);the Scientific Instrument Developing Project of the Chinese Academy of Sciences(YZ201668 to G.-Q.B.);the National Natural Science Foundation of China(91732304 to G.-Q.B.,91632303 to F.X.,91432305 to J.-N.Z.);the National Basic Research Program of China(2013CB835101 to G.-Q.B.);the Fundamental Research Funds for the Central Universities(WK6030000033 to H.W.)

摘　　要：The speed of high-resolution optical imaging has been a rate-limiting factor for meso-scale mapping of brain structures and functional circuits,which is of fundamental importance for neuroscience research.Here,we describe a new microscopy method of Volumetric Imaging with Synchronized on-the-fly-scan and Readout(VISo R)for high-throughput,high-quality brain mapping.Combining synchronized scanning beam illumination and oblique imaging over cleared tissue sections in smooth motion,the VISo R system effectively eliminates motion blur to obtain undistorted images.By continuously imaging moving samples without stopping,the system achieves high-speed 3D image acquisition of an entire mouse brain within1.5 hours,at a resolution capable of visualizing synaptic spines.A pipeline is developed for sample preparation,imaging,3D image reconstruction and quantification.Our approach is compatible with immunofluorescence methods,enabling flexible cell-type specific brain mapping and is readily scalable for large biological samples such as primate brains.Using this system,we examined behaviorally relevant whole-brain neuronal activation in 16 c-Fos-sh EGFP mice under resting or forced swimming conditions.Our results indicate the involvement of multiple subcortical areas in stress response.Intriguingly,neuronal activation in these areas exhibits striking individual variability among different animals,suggesting the necessity of sufficient cohort size for such studies.The speed of high-resolution optical imaging has been a rate-limiting factor for meso-scale mapping of brain structures and functional circuits, which is of fundamental importance for neuroscience research.Here, we describe a new microscopy method of Volumetric Imaging with Synchronized on-the-fly-scan and Readout(VISo R) for high-throughput, high-quality brain mapping.Combining synchronized scanning beam illumination and oblique imaging over cleared tissue sections in smooth motion, the VISo R system effectively eliminates motion blur to obtain undistorted images.By continuously imaging moving samples without stopping, the system achieves high-speed 3D image acquisition of an entire mouse brain within1.5 hours, at a resolution capable of visualizing synaptic spines.A pipeline is developed for sample preparation, imaging, 3D image reconstruction and quantification.Our approach is compatible with immunofluorescence methods, enabling flexible cell-type specific brain mapping and is readily scalable for large biological samples such as primate brains.Using this system, we examined behaviorally relevant whole-brain neuronal activation in 16 c-Fos-sh EGFP mice under resting or forced swimming conditions.Our results indicate the involvement of multiple subcortical areas in stress response.Intriguingly, neuronal activation in these areas exhibits striking individual variability among different animals, suggesting the necessity of sufficient cohort size for such studies.

关 键 词：fluorescence microscopy brain MAPPING tissue CLEARING IMMUNOSTAINING activity TRACE MAPPING 

分 类 号：R318[医药卫生—生物医学工程] TP391.41[医药卫生—基础医学]