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

X射线显微镜分析细胞内部结构和分子运输

• 批准号: 07680778
• 负责人: KIHARA Hiroshi
• 金额: $ 1.41万
• 依托单位: Kansai Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07680778/
• 关键词: x-ray microscopy; zone plate; synchrotron radiation; intracellular structure; endocytosis; immunogold; 結像型軟X線顕微鏡; シンクロトロン放射光; ナノイメージング; UVSOR; in vivo観察; ラベリング; 冷却CCDカメラ; in vivo 細胞観察; ホヤの血液細胞

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.

本研究的目的是观察细胞的细胞内结构，并寻找特定位点的特定标记技术，以供X射线显微镜监测。本研究使用的X射线显微镜首先设置在UVSOR，然后转移到RITS SR中心。目前的部分工作是在BESSY（柏林）完成的。1）X 射线站的建设。我们一直在通过以下方式改进 X 射线站： (1) 用光学显微镜和X射线显微镜观察同一标本。 (2) 将试样放置在空气中。探测器改为冷却CCD以提高其量子效率。有了这个系统；我们观察了硅藻、海鞘血细胞、衣藻、肌纤维、肿瘤细胞、原生质体和染色体等湿态和干态的生物样本。2）通过特定的标记技术分析细胞内结构和细胞内运输。关于这个主题，我们做了 3 项研究。 (1)通过用Au标记的抗体对微管进行染色，我们观察到了细胞骨架。 (2)COS细胞用高锰酸钾染色并在湿态下观察。 (3)通过X射线显微镜分析内吞作用。所有这三项研究都表明，特异性标记对于阐明湿态下的细胞内结构及其相关功能非常有用。

项目成果

N.Watanabe: "Imaging soft x-ray microscopy with zone plates,in parallel use of optical microscope for wet bio-specimens in air at UVSOR." in "X-ray Microscopy V(G.Shmahl et al.,ed)",Springer-Verlag,Berlin. (inpress).

N.Watanabe：“使用波带板对软 X 射线显微镜进行成像，同时使用光学显微镜在 UVSOR 下对空气中的湿生物样本进行成像。”

A.Hirai: "Soft x-ray imaging microscope with zone plates at RITS SR center." Rits SR center Activity Report 1996. (inpress).

A.Hirai：“RITS SR 中心配备波带板的软 X 射线成像显微镜。”

H.Kihara: "Biospecimens imaging with soft x-ray microscope." Rits SR center Activity Report 1996. (inpress).

H.Kihara：“使用软 X 射线显微镜进行生物样本成像。”

A.Hirai: "Measurement of quantum efficiency of the CCD camera systems." UVSOR activity report. 1996. 240-241 (1997)

A.Hirai：“CCD 相机系统量子效率的测量。”

N.Watanabe: "Imaging x-ray microscope with zone plates and its application to biological specimens at UVSOR." Zoological Studies. 34. 217-219 (1995)

N.Watanabe：“带有波带板的 X 射线成像显微镜及其在 UVSOR 生物样本中的应用。”