Iwabuchii Shinichiro, Mori Takuya, Ogawa Kenichi, Sato Kaori, Saito Masato, Morita Yasutaka, Ushiki Tatsuo, Tamiya Eiichi
School of Materials Science, Japan Advanced Institute of Science and Technology, Tatsunokuchi, Ishikawa, Japan.
Arch Histol Cytol. 2002 Dec;65(5):473-9. doi: 10.1679/aohc.65.473.
The present study was performed to introduce a novel chromosome dissection method employing atomic force microscopy (AFM) in a dynamic force mode for the chemical or molecular biological analysis of tiny chromosomal fragments. After AFM observation of human chromosomes prepared for light microscopy, a region of interest was dissected by increasing the loading force in a series of single-line scans of the target portion by controlling it with the amplitude reference of the tip in a dynamic force mode. The marker gene of the nucleolar organizing region (NOR) was amplified by our designed primers for 5.8S ribosomal DNA. After the dissection, topographic profiles in the section were then obtained with a carbon nanotube (CNT) probe in ambient condition. These results are discussed in relation to a fundamental technology for chromosomal analysis.
本研究旨在引入一种新颖的染色体切割方法,该方法采用原子力显微镜(AFM)在动态力模式下对微小染色体片段进行化学或分子生物学分析。在用光学显微镜制备的人类染色体进行AFM观察后,通过在动态力模式下以针尖的振幅参考来控制目标部分的一系列单行扫描中的加载力,从而切割感兴趣的区域。用我们设计的用于5.8S核糖体DNA的引物扩增核仁组织区(NOR)的标记基因。切割后,在环境条件下用碳纳米管(CNT)探针获取切片中的形貌轮廓。结合染色体分析的基础技术对这些结果进行了讨论。