2001 Fiscal Year Final Research Report Summary
Development of Atomic Force Microscope System Combined with Confocal Laser Scanning Microscope to Simultaneously Measure Mechanical Stiffness and Observe Microstructure of Cultured Cells
| Project/Area Number |
12480257
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biomedical engineering/Biological material science
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| Research Institution | Tohoku University |
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Principal Investigator |
SATO Masaaki Tohoku University, Graduate School of Engineering, Professor, 大学院・工学研究科, 教授 (30111371)
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| Co-Investigator(Kenkyū-buntansha) |
OHASHI Toshiro Tohoku University, Graduate School of Engineering, Research Associate, 大学院・工学研究科, 助手 (30270812)
MATSUMOTO Takeo Tohoku University, Graduate School of Engineering, Associate Professor, 大学院・工学研究科, 助教授 (30209639)
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| Project Period (FY) |
2000 – 2001
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| Keywords | Biomechanics / Shear stress / Endothelial Cell / Actin filament / GFP / FAT / Integrin / Finite element method |
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| Research Abstract |
Under flow condition endothelial cells are elongated and oriented to flow direction depending upon the level of shear stress and the duration of exposure. It is well known that cytoskeletal components, especially F-actin filaments, are playing important roles in the process of adaptation of cell shape to mechanical environment. In this project We have mainly focued two topics and obtained the results shown below.
1. Dynamic behavior of actin filament in living cells
We observed dynamic behavior of actin filaments in living cultured endothelial cells during exposure to shear stress. To do this, a vector of a green fluorescent protein (GFP)/actin fusion was introduced into cells using a lipofectoamine. Dynamic behavior of actin filament in an endothelial cell after introducing cytochalasin D was observed through CCD camera under an inverted fluorescent microscope. In different experiments, the endothelial cells were exposed to fluid shear stress of 2 Pa in a parallel plate flow chamber and
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the cytoskeletal structure was observed. Actin filaments located first at peripheral regions seemed to change the alignment after flow exposure and aligned to a long axis of the cell, which was not oriented to the flow direction yet, but already elongated.
2. Stress analysis in a cell exposed to shear stress
A fluid flow-structural analysis, i.e. a coupled field analysis, was performed to simulate three dimensional stress distribution in endothelial cells exposed to shear stress. The three-dimensional finite element model was generated on the basis of the cell surface geometry measured by an atomic force microscopy. The model consisted of a fluid element and a solid element representing the flow field and the endothelial cells, respectively. Analytical results on stress distribution in the cell showed that high compressive stress appeared both in the upstream side and the downstream side. These results may indicate that the stress distributions in the cells have close correlation with the F-actin distributions. Less
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