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

这个项目的目标是开发一种新类型的低温显微镜，用于观察冻结状态下的详细结构的细胞。对于光学系统来说，一束他的激光束已经完成了。激光束被引入到一个尼康Diaphoto TMD光学显微镜中，通过一个半反射镜光学玻璃纤维电缆和发光细胞的特性。辐射细胞的图像是通过光学眼睛或SIT视频摄像机通过图像增强器获得的。当激光束是由光学玻璃纤维电缆引入的时，该单元图像被严重地分散到斑点图案上。为了消除斑点模式，光学电缆在一个恒定的循环中随机振动。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 res\heater on which specimens are placed。蒸汽化的液体硝酸盐通过气体流量控制器系统流向冷阶段。该特殊温度已被测量为一个100微米铜-康斯坦丁热双，位于观察区中心。通过从可编程电压发生器引入一个小电流生成器来冷却和增强单元格的冷却和增强。温度的准确性在冷却和升温的范围内处于+0.1 ° C.通过使用这个低温显微镜，我们可以详细观察植物细胞在冰冻周期内的结构变化。在冷冻期间-原型塑料的循环，特征性的变化被观察到了两个等离子体膜和Tonoplasts上，提出了关于冷冻损伤的机制的新信息。

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.