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| 1 | 前沿生物医学电子显微技术的发展态势与战略分析显示文摘电子显微成像技术(简称电镜)因其超高分辨能力,成为人类认识微观世界的最主要技术手段之一,并广泛应用于生物医学研究领域.随着物理、生物、信息等多学科技术的发展和融合,电子显微镜在分辨率、成像尺度和自动化等多个方面不断取得突破,特别是冷冻制样技术和直接电子探测相机的发展使得冷冻电镜技术能够直接解析生物大分子的原子分辨率三维结构.另一方面,对生命活动过程的研究往往跨越多个时空尺度,甚至需要追踪生物事件发生的动态过程,这就要求研究手段在分辨率、原位成像、动态、多尺度、多模态、高特异性和高通量等特性上不断拓展.针对这些需求,生物医学电镜及相关技术的主要发展方向包括:(ⅰ)高分辨冷冻电镜技术,通过更精确的三维重构理论和电子光学成像技术进一步提高信噪比和分辨率,实现对更大、更小生物分子的原子分辨率结构解析;(ⅱ)基于冷冻电子断层三维重构的多模态成像技术,通过更优化的冷冻制样减薄技术、亚区域三维重建技术等,实现对组织与细胞中蛋白质等大分子复合物的结构与相互作用的原位解析,并通过与光学成像的动态性与特异性的有效整合,实现结构与功能研究的结合;(ⅲ)大尺度三维重建的体电子显微成像技术,通过优化样品制备、成像及分析,实现整个流程的高通量系统集成,从而进行脑联结组等生物结构的图谱重建;(ⅳ)具有飞秒量级时间分辨的超快电镜技术,通过进一步发展脉冲电子源、液相样品室等,将为蛋白质分子异构、生物组织与纳米材料的物理化学相互作用等动态过程研究提供新的观测工具;(ⅴ)大数据存储和处理技术与基于人工智能的图像处理方法,将进一步推动海量电镜数据的快速、高效、自动化的处理与分析.本文重点评述了以上技术方向的发展趋势、所面临的瓶颈问题以及可能的突破点,并展望了其在未来推动生物医学基础研究和转化应用的前景. | 陶长路 张兴 韩华 梁文锡 赵经纬 毕国强 | 2020 | 中国科学:生命科学2020，50，11： | 4 |
| 2 | HSV-1 H129-Derived Anterograde Neural Circuit Tracers:Improvements, Production, and Applications显示文摘Anterograde viral tracers are powerful and essential tools for dissecting the output targets of a brain region of interest. They have been developed from herpes simplex virus 1(HSV-1) strain H129(H129), and have been successfully applied to map diverse neural circuits.Initially, the anterograde polysynaptic tracer H129-G4 was used by many groups. We then developed the first monosynaptic tracer, H129-dTK-tdT, which was highly successful, yet improvements are needed. Now, by inserting another tdTomato expression cassette into the H129-dTK-tdT genome, we have created H129-dTK-T2, an updated version of H129-dTK-tdT that has improved labeling intensity. To help scientists produce and apply our H129-derived viral tracers, here we provide the protocol describing our detailed and standardized procedures. Commonly-encountered technical problems and their solutions are also discussed in detail. Broadly, the dissemination of this protocol will greatly support scientists to apply these viral tracers on a large scale. | Hong Yang Feng Xiong Yi-Ge Song Hai-Fei Jiang Hai-Bin Qin Jing Zhou Sha Lu Steven FGrieco Xiangmin Xu Wen-Bo Zeng Fei Zhao Min-Hua Luo | 2021 | Neuroscience Bulletin2021，37，5： | 4 |
| 3 | 整体器官的光透明成像方法综述显示文摘现代光学成像技术与荧光标记技术不断发展,为高分辨地获取生物组织三维结构信息提供了重要的工具。然而,大多数生物组织具有不透明特性,限制了光在组织中的穿透深度,进而限制了光学成像技术在大组织或整体器官成像中的应用。近年兴起的组织光透明技术通过多种物理、化学手段降低组织对光的衰减,增加光穿透深度,从而提高光学成像的成像深度与成像质量,为整体组织器官的三维成像提供了全新的思路。本文从离体组织光透明方法、大组织器官标记方法、三维整体成像技术三个方面,对整体器官的光透明成像方法进行综述。 | 俞婷婷 朱 | 2020 | 中国激光2020，47，2： | 3 |
| 4 | Miniature Fluorescence Microscopy for Imaging Brain Activity in Freely-Behaving Animals显示文摘An ultimate goal of neuroscience is to decipher the principles underlying neuronal information processing at the molecular,cellular,circuit,and system levels.The advent of miniature fluorescence microscopy has furthered the quest by visualizing brain activities and structural dynamics in animals engaged in self-determined behaviors.In this brief review,we summarize recent advances in miniature fluorescence microscopy for neuroscience,focusing mostly on two mainstream solutions-miniature single-photon microscopy,and miniature two-photon microscopy.We discuss their technical advantages and limitations as well as unmet challenges for future improvement.Examples of preliminary applications are also presented to reflect on a new trend of brain imaging in experimental paradigms involving body movements,long and complex protocols,and even disease progression and aging. | Shiyuan Chen Ziehen Wang Dong Zhang Aiming Wang Liangyi Chen Heping Cheng Runlong Wu | 2020 | Neuroscience Bulletin2020，36，10： | 2 |
| 5 | Whole-brain Optical Imaging:A Powerful Tool for Precise Brain Mapping at the Mesoscopic Level显示文摘The mammalian brain is a highly complex network that consists of millions to billions of densely-interconnected neurons.Precise dissection of neural circuits at the mesoscopic level can provide important structural information for understanding the brain.Optical approaches can achieve submicron lateral resolution and achieve“optical sectioning”by a variety of means,which has the natural advantage of allowing the observation of neural circuits at the mesoscopic level.Automated whole-brain optical imaging methods based on tissue clearing or histological sectioning surpass the limitation of optical imaging depth in biological tissues and can provide delicate structural information in a large volume of tissues.Combined with various fluorescent labeling techniques,whole-brain optical imaging methods have shown great potential in the brain-wide quantitative profiling of cells,circuits,and blood vessels.In this review,we summarize the principles and implementations of various whole-brain optical imaging methods and provide some concepts regarding their future development. | Tao Jiang Hui Gong Jing Yuan | 2023 | Neuroscience Bulletin2023，39，12： | 0 |
| 6 | 全脑显微光学成像显示文摘全脑介观神经联接研究是解析脑认知功能的神经输入输出环路结构基础、普查基因表达与细胞类型,以及绘制全景立体脑图谱的科学前沿。光学成像方法在横向方向能够达到亚微米的分辨率,并可通过多种手段实现“光学切片”的效果,具备在介观水平观测神经环路的天然优势。基于组织透明或机械切削的自动化全脑显微光学成像方法,突破了光学成像在生物组织中成像深度的限制,具有在大范围内提供介观水平精细观察的技术优势。结合各类生物样本荧光标记技术,全脑显微光学成像方法在神经环路的结构和功能的研究方面有着巨大潜力,已成为剖析全脑神经及血管网络的最佳方式。为了更全面地了解和认识这种有力的工具,总结了近年来发展的各类全脑显微光学成像方法,并展望了未来的技术发展。 | 江涛 龚辉 骆清铭 袁菁 | 2023 | 中国激光2023，50，3： | 0 |
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