Analysis on the internal structure and transport of molecules of the cell by x-ray microscope

本研究的目的是观察细胞的细胞内结构，探索用于X射线显微镜监测的特定部位的特定标记技术。本研究使用的X射线显微镜首先在UVSOR安装，然后转移到RITS SR中心。目前的部分工作是在BESSY(柏林)1)建造X射线站。我们一直在改进X射线站：(1)用光学显微镜和X射线显微镜观察相同的标本。(2)在空气中放置标本。为了提高探测器的量子效率，探测器改用了冷却的电荷耦合器件。利用该系统，我们在干湿状态下观察到了硅藻、海鞘血细胞、脊索藻、肌纤维、肿瘤细胞、原生质体和染色体等多种生物。2)用特殊标记技术对细胞内结构和细胞内运输进行了分析。在这个问题上，我们已经做了3项研究。(1)用Au标记的抗体对微管进行染色，观察到细胞角质形成。(2)COS细胞高锰酸钾染色，湿法观察。(3)X线显微镜下细胞吞噬功能分析。这三项研究都表明，在湿状态下，特异性标记对于阐明细胞内结构及其相关功能是非常有用的。

The purpose of the present research is to observe intracellular structure of the cells and to search specific labeling techniques of the specific site for monitoring by x-ray microscopy.The x-ray microscope used in the present research is firstly set up at UVSOR,and then moved to RITS SR center. A part of the present works were done at BESSY (Berlin).1) Construction of the x-ray station. We have been improving the x-ray station by ; (1) To observe the same specimens by the light microscope as well as by x-ray microscope. (2) To set specimens in the air. The detector was changed to cooled CCD to improve its quantum efficiecy. With this syustem ; we have observed many biospecimens such as diatoms, blood cells of ascidian, clamydomonas, muscle fibers, tumor cells, protoplasts and chromosomes both in wet and dried states.2) Analysis on the intracellular structure and intracellular transport by specific lableing techniques. On this subject, we have done 3 studies. (1) By staining microtubles with Au-labeled antiubodies, we observed cytoskeltons. (2) COS cells were stained by potassium permanganate and observed in wet state. (3) Analysis of endocytosis by x-ray microscopy. All of the three researches demonstrate that specific labeling is very useful in elucidating intracellular structure and its related functions in wet state.