Imaging of molecular and cellular functions by scanning probe microscopy

通过扫描探针显微镜对分子和细胞功能进行成像

• 批准号: 13470002
• 负责人: USHIKI Tatsuo
• 金额: $ 3.84万
• 依托单位: NIIGATA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470002/
• 关键词: scanning probe microscopy; atomic force microscopy biomolecules; cellular function; 細胞機能; 高分子観察; 走査プローブ顕微鏡; 高分子; 液中観察; 近接場光学顕微鏡

Scanning probe microscopes form a new family of microscopes, which scan a sharp probing tip over the sample surface, providing high resolution information about the surface characteristics of solid samples. This study was undertaken to apply these microscopes (including the atomic force microscope) to the observation of biological samples. For this purpose, we also investigated the preparation methods and observation techniques suitable for scanning probe microscopy. The obtained results are follows :1.We studied the techniques for observing macromolecules such ad the deoxyribonucleic acid and myosin at high resolution with the atomic force microscope. These samples are usually mounted on a fleshly-cleaved mica and observed with the microscope in an air environment. In this study, we applied an FM control system to the visualization of macromolecule in a vacuum environment, and succeeded in observing them at high resolution.2.We investigated the methods for observing biological samples (e.g., chromosomes) with the atomic force microscope in a liquid environment. While a contact mode is generally used for measuring soft samples in liquid, we investigate the method for operating the atomic force microscope in a dynamic mode. By adjusting adequately the operation parameters such as resonance frequency, amplitude and vibration voltage, we succeeded in obtaining clear images of chromosomes in liquid using a dynamic mode. Q-control technique for dynamic mode imaging was expected to be useful for the stable visualization of chromosomes without any fixation in a liquid environment.

扫描探针显微镜形成了一个新的显微镜家族，它在样品表面上扫描一个尖锐的探针尖端，提供有关固体样品表面特性的高分辨率信息。本研究旨在将这些显微镜（包括原子力显微镜）应用于生物样品的观察。为此，我们还研究了适用于扫描探针显微镜的制备方法和观察技术。1.研究了用原子力显微镜对脱氧核糖核酸和肌球蛋白等大分子进行高分辨观察的技术。这些样品通常安装在一个肉裂云母和观察显微镜在空气中的环境。在本研究中，我们将FM控制系统应用于真空环境中大分子的可视化，并成功地在高分辨率下观察它们。2.我们研究了观察生物样品的方法（例如，染色体）与原子力显微镜在液体环境中。虽然接触模式通常用于测量液体中的软样品，但我们研究了在动态模式下操作原子力显微镜的方法。通过适当调节共振频率、振幅和振动电压等操作参数，我们成功地利用动态模式获得了液体中染色体的清晰图像。动态模式成像的Q-控制技术有望用于在液体环境中稳定显示染色体而无需任何固定。

项目成果

T.Yoshino, S.Sugiyama, S.Hagiwara, T.Ushiki, T.Ohtani: "Simultaneous collection of topographic and fluorescent images of barley chromosomesby scanning near-field optical/atomic force microscopy"Japanese Society of Electron Microscopy. 51(3). 199-203 (2002

T.Yoshino、S.Sugiyama、S.Hagiwara、T.Ushiki、T.Ohtani：“通过扫描近场光学/原子力显微镜同时收集大麦染色体的形貌和荧光图像”日本电子显微镜学会。

Tatsuo Ushiki: "Collagen Fibers, Reticular Fibers and Elastic Fibers. A Comprehensive Understanding from a Morphological Viewpoint"Archives of Histology and Cytology. 65-2. 109-124 (2002)

Tatsuo Ushiki：“胶原纤维、网状纤维和弹性纤维。从形态学角度的全面理解”组织学和细胞学档案。

O.Hoshi, T.Ushiki: "Three-dimensional Structure of G-banded Human Metaphase Chromosomes Observed by Atomic Force Microscopy"Archives of Histology and Cytology. 64-5. 475-482 (2001)

O.Hoshi，T.Ushiki：“原子力显微镜观察到的 G 带人类中期染色体的三维结构”组织学和细胞学档案。

T.Ushiki, O.Hoshi, K.Iwai, E.Kimura, M.Shigeno: "The Structure of Human Metaphase Chromosomes : Its Histological Perspective and New Horizons by Atomic Force Microscopy"Archives of Histology and Cytology. 65-5. 377-390 (2002)

T.Ushiki、O.Hoshi、K.Iwai、E.Kimura、M.Shigeno：“人类中期染色体的结构：组织学视角和原子力显微镜的新视野”组织学和细胞学档案。

Ushiki T, Hoshi O, Iwai K, Kimura E, Shigeno M.: "The structure of human metaphase chromosomes : its histological perspective and new horizons by atomic force microscopy."Arch Histol Cytol.. 65-5. 377-390 (2002)

Ushiki T、Hoshi O、Iwai K、Kimura E、Shigeno M.：“人类中期染色体的结构：原子力显微镜的组织学视角和新视野。”Arch Histol Cytol.. 65-5。