一种高通量大尺度生物样品电镜三维重构方法的研究

项目名称： 一种高通量大尺度生物样品电镜三维重构方法的研究

项目编号： No.31501160

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 遗传学与生物信息学、细胞生物学

项目作者： 李喜霞

作者单位： 中国科学院生物物理研究所

项目金额： 20万元

中文摘要： 随着对细胞生物学、神经生物学的深入研究，人们越来越需要获得大尺度生物样品的三维结构信息，而这离不开体电子显微学技术的发展。目前体电子显微学技术主要有：基于透射电镜成像的连续超薄切片策略，和基于扫描电镜成像的系列块表面、原位离子束切割以及刚起步尚需完善还未广泛应用的自动化带式收集超薄切片机三种策略。在过去的两年工作中，我们已率先成功研制出第一代连续切片自动收集装置，并对大尺度三维重构新方法进行了初步探讨。在此工作基础上，本项目拟选取线虫作为研究对象，进一步优化并发展以连续切片自动化收集和扫描电镜自动化成像为核心技术的高通量大尺度生物样品三维重构的新方法，并将构建一套完整的线虫三维结构数据库。项目研究内容包括：（1）适合连续切片自动收集和扫描电镜成像的样品制备；（2）高通量连续切片的自动化收集；（3）高通量扫描电镜自动化成像技术的实现；（4）大尺度三维重构及结构分析。

中文关键词： 连续超薄切片；自动化收集；扫描电子显微镜；三维重构；秀丽隐杆线虫

英文摘要： With the in-depth development of life science, the 3D structure with high resolution of large scale biological samples become more and more important. It depended on the development of volume electron microscopy (EM). Currently, there are four volume EM strategys:1)Serial section transmission electron microscopy(ssTEM); 2) Serial Block-Face scanning electron microscopy(SBF-SEM);3) Focused Ion Beam SEM(FIB-SEM);4)Automatic Tape- Collecting Ultramicrotome SEM(ATUM-SEM). And the ATUM-SEM technology has not been widely used because it still need to be improved in its start-up phase. In the past two years，we have successfully developed the first generation of serial ultrathin section automated collection device, and carried out the research on a new method of large-scale 3D reconstruction. Based on the past work，we intend to further optimize and develop the new high throughout 3D reconstruction method by selecting Caenorhabditis elegans as the sample. For this project, the serial ultrathin sections automated collection technology and the scanning electron microscopy (SEM) automated imaging technology are the core technologies. Meanwhile, a database for C.elegans 3D ultrastructure will also be established with this method. The research content includes:(1) the preparation of the suitable resin embedded block; (2) the automated collection of high throughout serial sections; (3) the automated collection of high throughout SEM imaging; (4) the large-scale 3D reconstruction and analysis.

英文关键词： serial ultrathin section;automated collection;scanning electron microscopy ;3D reconstruction;Caenorhabditis elegans