Development of automatic electron microscope for tomography and application to cell biology

自动断层扫描电子显微镜的研制及其在细胞生物学中的应用

• 批准号: 17500305
• 负责人: TAKAOKA Akio
• 金额: $ 1.92万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17500305/
• 关键词: Electron tomography; Automatic electron microscope; Automatic alignment; 3D observation of cells; Automatic CT; 細胞診断; トモグラフィー; 電子顕微鏡; 立体解析; 自動化; ソフト開発

We have developed an automatic electron microscope for tomogaraphy on the basis of a Hitachi H-9500 EM whose acceleration voltage is 200 or 300 kV. This EM has following features ; (a)the pole piece designed for high angle tilt up to 70deg, (b)the eucentric goniometer with five adjustable axes, (c)the high sensitivity CCD camera with 2k x 2k pixels (TemCam-F224S, TVIPS) and (d)the automatic image acquisition for a tilt-series. The main points of the development are the interface between the EM and the camera, and the software EM-menu of TVIPS enables us to operate the auto-tracking and the auto-focusing for the EM image.After acquisition of the tilt-series EM images, the tomogram is reconstructed with the software IMOD developed at Colorado University. Then, we added a function that the arraignment of tilt-series images carries out automatically without fiducial markers such as artificial gold particles. That is, we developed new software named "CA markers" that can choose the feature points and truck the each point among the tilt-series images.With the system and software, we reconstructed 3D images of a few samples such as microvillus of duodenum, HIV invaded into cell membrane and nuclear of malignant cells.

我们在加速电压为200或300 kV的日立H-9500 EM的基础上开发了一种用于断层摄影的自动电子显微镜。该EM具有以下特点：（a）极片设计用于高达70度的高角度倾斜，（B）具有五个可调轴的同心测角仪，（c）具有2k x 2k像素的高灵敏度CCD相机（TemCam-F224 S，TVIPS）和（d）倾斜系列的自动图像采集。重点是EM和摄像机的接口，TVIPS的EM菜单软件实现了EM图像的自动跟踪和自动聚焦，采集了倾斜序列的EM图像后，用科罗拉多大学的IMOD软件重建了断层图像。然后，我们添加了一个功能，即倾斜系列图像的排列自动执行，而无需人工金颗粒等基准标记。利用该系统和软件，我们对十二指肠微绒毛、HIV侵入细胞膜、恶性细胞核等几个样本进行了三维重建。

项目成果

Microscopic tomography with ultra-HVEM and application

超 HVEM 显微断层扫描及其应用

• 发表时间: 2007
• 期刊: Ultramicroscopy (submitted)
• 作者: Akio Takaoka;T.Hasegawa;K.Yoshida;H.Mori
• 通讯作者: H.Mori

Microscopic tomography with ultra-UHEM and application

超 UHEM 显微断层扫描及其应用

• 发表时间: 2007
• 期刊: Ultramicroscopy (submitted)
• 作者: Akio Takaoka;T.Hasegawa;K.Yoshida;H.Mori
• 通讯作者: H.Mori

超高圧電子顕微鏡トモグラフィーによる立体観察技術〜病理組織から半導体デバイスまで〜

利用超高压电子显微镜断层扫描的三维观察技术～从病理组织到半导体器件～

• 发表时间: 2005
• 期刊: 顕微鏡学会関東支部第30回講演会予稿集
• 作者: A.Takaoka;H.Mori;鷹岡昭夫
• 通讯作者: 鷹岡昭夫

Nano-observation of HIV entry process by electron microscopic tomography

电子显微镜断层扫描对HIV侵入过程的纳米观察

• 发表时间: 2006
• 期刊: Proc. 16^<th> International Microscopy Congress vol. 2
• 作者: Toshiyuki Goto;A. Takaoka
• 通讯作者: A. Takaoka

Feature of ultra-HVEM tomography and application

ultra-HVEM断层成像的特点及应用

• 发表时间: 2006
• 期刊: Proc. 4^<th> Int. Cong. on Electron Tomography Vol. 1
• 作者: Akio Takaoka;T.Hasegawa;K.Yoshida;H.Mori
• 通讯作者: H.Mori