Development of a laser optical cryomicroscope for the study of freeze-preservation of plant gene resources.

开发用于植物基因资源冷冻保存研究的激光光学冷冻显微镜。

• 批准号: 60840020
• 负责人: YOSHIDA Shizuo
• 金额: $ 15.36万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research
• 财政年份: 1985
• 资助国家: 日本
• 起止时间: 1985 至 1987
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-60840020/
• 关键词: Laser optical microscope; Cryomicroscope; Cell cooling system; Process of freeze-thawing cycle; freeze-preservation of gene resources; 凍結傷害の機構; 細胞形態; 植物遺伝子源; 凍結保存; 凍結傷害

The objective of this project is to develop a new type of cryomicroscope for the observation of detailed structure of cells under the frozen state. For the optical system, a He-Ne laser beam is equipped. The laser beam is introduced into a Nikon Diaphoto TMD optical microscope through an optical glass-fiber cable and illuminated cell specimens through a half mirror. The images of the irradiated cells are viewed through an optical eyepiece or a SIT video camera equipped with a image enhancer device. When the laser beam is introduced by the optical glass fiber cable, the cell images are severely disturbed due to the speckle patterns. To eliminate the speckle patterns, the optical cable is randomly vibrated at a constant cycle. The light microscope is also equipped with bright field, phase contrast, differential interference contrast optics, or fluorescein illustration system.Cooling and rewarming of cell specimens are effected by a "conduction heat transfer" stage consisting of a copper cold sink interfaced with a transparent electrical resistance heater on which specimens are placed. Vaporized liquid nitrogen flows to the cold stage through a gas-flow controller system. the specimen temperature is measured with a 100 um copper-constantan thermocouple positioned at the center of the view region. Cooling and rewarming of cell specimens are achieved by introducing a small current generated from programmable voltage generator. The temperature accuracy during cooling and rewarming is in the range of +0.1゜C.By using this cryomicroscope, we can observe detailed structural changes of plant cells during freeze-thaw cycle. During freeze-thaw cycle of protoplasts, characteristic changes were observed on both plasma membranes and tonoplasts, suggesting a new information on the mechanism of freezing injury of plant cells.

为了观察细胞的cetail结构，将HE-NE激光束装备到尼康透明度TMD光学显微镜中。电缆和通过半镜子的镜头来看，通过光学摄像机或配备了IIMAGE增强器设备的坐姿摄像机。 。放置样品的电阻加热器，通过液体液体氮流到冷阶段，用W冷却来测量样品温度。可编程的电压发生器。植物细胞冻结的机制。

项目成果

S.Etani ; S.Yoshida.: Plant & Cell Physiology. 28. 83-91 (1987)

埃塔尼；

菅原康剛・竹内正幸: 凍結及び乾燥研究会会誌. 33. 45-48 (1987)

Yasutaka Sukawara 和 Masayuki Takeuchi：《冷冻干燥研究会杂志》33. 45-48 (1987)。

Plant Physiology. 80-1. (1986)

植物生理学。

S.Yoshida;M.Uemura: Plant Physiology. 82. 807-812 (1986)

S.Yoshida；M.Uemura：植物生理学。

S.Yoshida.: "Resposes of the plasma membranes to freezing stress.In The Plant Plasma Membrane : Structure, Fuction and Molecular Biology" Springer Verlag, 26 (1988)

S.Yoshida.：“质膜对冷冻应激的反应。植物质膜：结构、功能和分子生物学”Springer Verlag，26 (1988)