Improvement of specimen preparation technique in atomic force microscope for the bacteriological use.

细菌学用原子力显微镜标本制备技术的改进。

• 批准号: 07557213
• 负责人: AMAKO Kazunobu
• 金额: $ 0.7万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 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-07557213/
• 关键词: AFM,; plasma polymerization; imaging; high resolution; filamentous phage; SPM; プラズマ重合; 線維状ファージ; グラム陰性石; ファージ; 細菌; 顕微鏡

A filamentous bacteriophage pf of Pseudomonas aeruginosa was examined by contact or noncontact mode of AFM.Purified phages were applied on clean cover glass and air dried. By contact mode the phage was able to be imaged by the first scan but the image was soon disappeared or dislocated in 2nd or 3rd scanning. The quality of the image in high magnification was well in this mode. By noncontact mode repeated imaging of the phage filament was capable but the quality of the image was less than that obtained by contact mode. After coating the specimens with a thin layr of plasma polymerized butane, even by the contact mode, repeated imaging became capable. The image quality was high in the initial a few imaging scans. The thin film of polymerized butane was so hard that the tip for imaging was soon worm out and resulted in the deterioration of the imaging quality.Improvement of the tip is important for high resolution imaging in biological specimen.

用AF法对铜绿假单胞菌的丝状噬菌体Pf进行了接触和非接触检测，将纯化后的噬菌体涂抹在干净的盖子上风干。在接触模式下，噬菌体在第一次扫描时就能成像，但在第二次或第三次扫描时，图像很快就消失或错位。在这种模式下，高倍率下的图像质量很好。在非接触模式下，噬菌体细丝可以重复成像，但图像质量不如接触模式。在样品上涂上一层薄薄的等离子体聚合丁烷后，即使是通过接触模式，重复成像也是可能的。在最初的几次成像扫描中，图像质量很高。由于聚合丁烷薄膜太硬，成像针尖很快就会蠕动起来，导致成像质量下降，改善针尖对生物样品的高分辨率成像具有重要意义。

项目成果

K.Amako 他: "Imaging of the surface structures of Epon thin sections created with glass krife and a Diamond Krife by the atomic force microsope" J.Electron microscopy. 42. 121-123 (1993)

K.Amako 等人：“通过原子力显微镜对使用玻璃刀和金刚石刀创建的 Epon 薄片的表面结构进行成像”J.Electron Microsope 42. 121-123 (1993)。

天児和暢・梅田昭子: "走査プローブ顕微鏡" 日本細菌学雑誌. 51. 689-696 (1996)

Kazunobu Amago 和 Akiko Umeda：“扫描探针显微镜”日本细菌学杂志 51. 689-696 (1996)。

A.Umeda 他: "Stabilization of Biological specmins by use of plasma poly-merijed hydi carbon film for imaging with an AFM" J.Electron Microscopy. 44. 408-409 (1995)

A.Umeda 等人：“通过使用等离子体聚合碳氢薄膜进行 AFM 成像来稳定生物样本”J.Electron Microscopy 44. 408-409 (1995)。

Umeda A.et al: "Stabilization of Biological specimens by use of plasma polymelized hydrocarbon film for imaging with an AFM" J.Electron microscopy. 44. 408-409 (1995)

Umeda A.等人：“使用等离子体聚合碳氢化合物薄膜通过 AFM 成像来稳定生物样本”J.电子显微镜。

K.Amako 他: "Imaging of the surface struicture of Epon thin section creatid Uth glass knif and a diamond kife by AFM" J.Electron Microscopy. 42. 121-123 (1993)

K.Amako 等人：“通过 AFM 对 Epon 薄切片 creatid Uth 玻璃刀和金刚石刀的表面结构进行成像”J.Electron Microscopy 42. 121-123 (1993)。