1.1)中国科学院生物物理研究所脑与认知科学国家重点实验室,北京 100101;2.2)中国科学院大学,北京 100049;3.3)上海交通大学医学院附属瑞金医院卢湾分院,上海 200020;4.4)中国科学院自动化研究所,北京 100190;5.5)中国科学院神经科学研究所神经科学国家重点实验室,中国科学院脑科学与智能技术卓越创新中心,上海 200031;6.6)上海脑科学与类脑研究中心,上海 201210;7.7)中国科学院上海微系统与信息技术研究所,上海 200050
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中国科学院战略性先导科技专项(B类)(XDB32010100)
1.1)State Key Laboratory of Brain and Cognitive Science, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China;2.2)University of Chinese Academy of Sciences, Beijing 100049, China;3.3)Ruijin Hospital, Luwan Branch, Shanghai Jiao Tong University School of Medicine, Shanghai 200020, China;4.4)Institute of Automation, Chinese Academy of Sciences, Beijing 100190, China;5.5)Institute of Neuroscience, State Key Laboratory of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai 200031, China;6.6)Shanghai Center for Brain Science and Brain-Inspired Intelligence Technology, Shanghai 201210, China;7.7)Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, China
the Strategic Priority Research Program of Chinese Academy of Science, Grant No.XDB32010100
三维电子显微成像(volume electron microscopy)是微观尺度脑联结组学(micro-brain connectomics)研究的主要研究手段.自动卷带式连续超薄切片-收片系统(automated tape-collecting ultramicrotome, ATUM)及扫描电镜序列成像系统的联合应用,是三维电镜成像中一种重要的技术手段.该方法利用卷带式收集超薄切片,形成完整的样品切片库,进而在后续成像中形成样品三维电镜成像数据库.我们根据ATUM系统镜外切片-收片、保留样本切片库等特点,对相应的生物样品制备方法进行改进,包括树脂配方及树脂渗透方式改良;神经元细胞膜辣根过氧化物酶稀疏化标记与样本切片库后染色联用——从而获得符合脑联结组学所需的高质量成像数据.
For micro-brain connectomic research, volume electron microscopy is a main method in which it plays an important role for the application of automated tape-collecting ultramicrotome (ATUM) system and array imaging with scanning electron microscopy together. ATUM system enables collecting a large amount of serial sections to be imaged with SEM to form a huge dataset. With regard to the nature of non-vacuum sectioning and sample reserving, we modified and improved the sample preparation process including resin recipe, HRP labelling and post-staining method in order to obtain dataset of high quality for connectomic research.
王圣雄,陈贞强,韩华,郭爱克.辣根过氧化物酶标记与样品后染色联用在微观脑联结组学中的应用[J].生物化学与生物物理进展,2020,47(10):1090-1096
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