Development of molecular anatomical technique of the cell by the scanning probe microscop
Grant-in-Aid for Scientific Research (A)
|Allocation Type||Single-year Grants|
General anatomy (including Histology/Embryology)
|Research Institution||Kitasato University School of Medicine|
YAMASHINA Shohei Kitasato Univ.School of Medicine, Professor, 医学部, 教授 (90013987)
|Project Period (FY)
1995 – 1997
Completed(Fiscal Year 1997)
|Budget Amount *help
¥15,900,000 (Direct Cost : ¥15,900,000)
Fiscal Year 1997 : ¥1,000,000 (Direct Cost : ¥1,000,000)
Fiscal Year 1996 : ¥1,100,000 (Direct Cost : ¥1,100,000)
Fiscal Year 1995 : ¥13,800,000 (Direct Cost : ¥13,800,000)
|Keywords||SPM / Cellular ultrastructure / Cellular membrane / Molecular anatomy / Freeze fracture / Immuno-histochemistry / Enzyme-histochemistry / Colloidal gold particle / 走査型プローブ顕微鏡 / 細胞構造 / 組織化学 / AFM / DFM / 凍結技法|
Cellular ultra stucture and identification of functional molecules was determined with a use of the present SPM,and following results were obtained.
(1) Uitra fine structure of cells as revealed by SPM
Comparative study of various sample preparation imdicated that the best resolution could be obtained by a combination of cyclic contact mode with ultra sharpen needle. According to this, cellular ultra structure was resolved at about 30nm resolution. Further sharpening of needle tip was feasible in order to power up the resolution.
(2) Application of freeze fractured and SDS treated sample.
Observation of freeze fractured and SDS treated samples were made to determine the true surface of the cell under the best condition stated above. As a result, irregular processes about 30nm in diameter could be visualized on the cellular true surface. These structure seemed to correspond with compounds of lipid and protein molecules that consisted of cellular membrane. It was expected to visualize more detailed image of the cell surface by this preparation, if resolution of the SPM could be raised.
(3) Identification of functional molecules by the SPM
Colloidal gold particle labelled by immunohistochemical method could be detected by their configuration, however, lateral diameter tended to appear slightly larger than expected amount due to lateral effect of needle tip. Not only gold particles but also lead phosphate, final reaction products of enzyme histochemical reaction could be detected with a use KFM that made it possible to detect the difference in surface electrical potential. The present KFM has several problems of slow response to local potential differences.
Research Output (9results)