Improvement and application of high-vacuum evaporater with condensed-light beam heater for deep-etching replication of TEM specimen

聚光束加热器高真空蒸发器深蚀复制TEM样品的改进及应用

• 批准号: 02559005
• 负责人: TONOSAKI Akira
• 金额: $ 11.39万
• 依托单位: Yamagata University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02559005/
• 关键词: Clean-vacuum evaporater; Freeze-fracturing replica; Deep-etching replica; Surface-sublimation by condensed-light beam; Biological membranes; 真空蒸着装置; ディ-プ・エッチング法; 網膜; 光照射エッチング

A high-vacuum evaporater has been proposed and completed in the form of JFD-9010S (JOEL) by this project. It is characterized by 'clean vecuum' with a turbomolecular and a cryo-absorption pump, keeping out the traditional oil-diffusion pump. For facilitating the deep-etching or sublimation process on the fractered surface of the specimen maintained at the low-temperature stage, a condensed-light heating device has been adopted. Using this innovated device, adequate deep-etching is obtained after 40-60 min irradiation during which the specimen is maintained at -110 to -120ﾟC, limiting the sublimation to the very surface to be replicated.Application to the biological membrane structure, including of photoreceptor cells, halobacteria, and diabetic neural fibers, is going on. The final data will be quatitatively analyzed with the special soft-ware arranged for counting the intramembrane particles and other deep-etching fine structures on TEM pictures projected to CRT.

本项目提出并完成了JFD-9010S（JOEL）高真空蒸发器。它的特点是具有涡轮分子泵和低温吸收泵的“清洁真空”，排除了传统的油扩散泵。为了便于在低温阶段对样品的破碎表面进行深蚀刻或升华过程，采用了聚光加热装置。使用这种创新设备，在40-60分钟的照射后获得充分的深度蚀刻，在此期间将样本保持在-110至-120°C，将升华限制在要复制的表面。目前正在将其应用于生物膜结构，包括感光细胞、嗜盐细菌和糖尿病神经纤维。最终数据将使用专门的软件进行定量分析，该软件用于对投影到 CRT 的 TEM 图片上的膜内颗粒和其他深蚀刻精细结构进行计数。

项目成果

外崎 昭: "特集 光受容膜によせて" 細胞. 22. 2-2 (1990)

Akira Tonosaki：“光感受器膜的特殊功能”细胞。22. 2-2 (1990)

Hiroshi Watanabe: "Intramembrane particles in the postsynaptic membranes of the S-, F-, and C-type synapses by freeze-fracturing and deepetching studies on the Xenopus spinal cord." Adv. Exp. Med. Biol.287. 249-254 (1991)

Hiroshi Watanabe：“通过对非洲爪蟾脊髓的冷冻断裂和深蚀刻研究，发现了 S、F 和 C 型突触后膜中的膜内颗粒。”

Hiroshi Watanabe: "Nicotinic and muscarinic postsynaptic membranes:Ultrathin-section and freeze-fracture studies." J.Autonom.Nerv.Sys.33. 144-145 (1991)

Hiroshi Watanabe：“烟碱和毒蕈碱突触后膜：超薄切片和冷冻断裂研究。”

渡辺 皓: "「電子顕微鏡チャ-トマニュアル」レプリカ法:解説" 二重作 豊、安達 公一、朝倉 健太郎(編者)、医学出版センタ-, (1992)

渡边晃：《电子显微镜图表手册》复制方法：评论，Yutaka Yutaka，Koichi Adachi，Kentaro Asakura（编辑），医学出版中心，（1992）

Masahiro Hara: "Fine structure and volumetric changes of lamprey photoreceptor cells during light and dark perilds." Cell Tiss Res.259. 33-41 (1990)

Masahiro Hara：“七鳃鳗感光细胞在光照和黑暗时期的精细结构和体积变化。”