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Development of Atomic Force Microscope System Combined with Confocal Laser Scanning Microscope to Simultaneously Measure Mechanical Stiffness and Observe Microstructure of Cultured Cells

Research Project

Project/Area Number 10558126
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Biomedical engineering/Biological material science
Research InstitutionTOHOKU UNIVERSITY

Principal Investigator

SATO Masaaki  Department of Mechatronics and Precision Engineering, Tohoku University, Professor, 大学院・工学研究科, 教授 (30111371)

Co-Investigator(Kenkyū-buntansha) HAYASHI Yoshiaki  Product Development Department, Olympus Optical Co., Ltd., Research Manager, 光学機器開発部, 課長(研究職)
OHASHI Toshiro  Department of Mechatronics and Precision Engineering, Tohoku University, Research Assistant, 大学院・工学研究科, 助手 (30270812)
MATSUMOTO Takeo  Department of Mechatronics and Precision Engineering, Tohoku University, Associate Professor, 大学院・工学研究科, 助教授 (30209639)
KATAOKA Noriyuki  Department of Medical Engineering, Kawasaki College of Allied Health Professions, Assistant Professor, 医用電子技術科, 講師 (20250681)
Project Period (FY) 1998 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥12,500,000 (Direct Cost: ¥12,500,000)
Fiscal Year 1999: ¥4,600,000 (Direct Cost: ¥4,600,000)
Fiscal Year 1998: ¥7,900,000 (Direct Cost: ¥7,900,000)
KeywordsAtomic Force Microscope / Confocal Laser Scanning Microscope / Endothelial Cell / Mechanical Property / Actin Filament / Shear Stress / Finite Element Method
Research Abstract

The atomic force microscope (AFM) system was originally developed in combination with an inverted confocal laser scanning microscope to simultaneously measure mechanical stiffness and to observe microstructure of cultured cell. To detect an indentation depth of the cantilever, a cantilever moving system was introduced. A special specimen holder was made to hold a commercially available culture dish. The movements of the cantilever and the specimen holder were controlled by a personal computer. This system was applied to statically cultured and shear stress exposed endothelial cells, and the following results were obtained.
1. Observation of actin filaments and measurement of three dimensional configuration were performed for a fixed endothelial cell.
2. Input and output (I/O) ports of culture medium were newly designed to use a culture dish for flow exposure experiments. Morphology and mechanical properties of cultured endothelial cells were measured using the AFM system. Endothelial cells cultured at static condition had a polygonal shape and more soft mechanical properties around a nucleus than those at peripheral regions. The stiffness of the endothelial cells exposed to shear stress of 2 Pa became higher with the duration time of exposure. Cell shape became elongated to the flow direction and the location of a nucleus moved to downstream side by shear flow.
3. A fluid flow-structural analysis was performed. The three-dimensional finite element model was generated on the basis of the cell surface geometry measured by the AFM. 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 mainly in the upstream side. This result may indicate that the stress distributions in the cells have close correlation with the F-actin distributions.

Report

(3 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • Research Products

    (33 results)

All Other

All Publications (33 results)

  • [Publications] 片岡則之: "力学的刺激と内皮細胞骨格構造の変化"日本バイオレオロジー学会誌. 12. 234-246 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 片岡則之: "せん断応力負荷の初期過程における培養内皮細胞の形態およびF-アクチンフィラメントの変化"日本機械学会論文集(B編). 64. 1801-1808 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 片岡則之: "流れの方向変化が培養内皮細胞の形態に及ぼす影響-流れの方向変化の時間間隔の影響-"医用電子と生体工学. 36. 302-308 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 武田浩一郎: "培養動脈組織片を用いた内皮細胞の流れに対する形態的反応の検討"医用電子と生体工学. 36. 293-301 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kataoka N.: "The effect of flow direction on the morphological responses of cultured bovine aortic endothelial cells"Medical & Biological Engineering & Computing. 36. 122-128 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kataoka N.: "The morphological responses of cultured bovine aortic endothelial cells to fluid-imposed shear stress under sparse and colony conditions"JSME International Journal Series C. 41. 76-82 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kataoka N.: "How do flow directions affect the morphology of cultured endothelial cells? -In relation to the blood flow condition at atherosclerotic regions-"Proc. the 10th Int. Conf. on Mechanics in Medicine and Biology. 133-136 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Kataoka N.: "Effects of cell-cell and cell-substrate contacts on the response of cultured endothelial cells to fluid shear stress"Proc. of the 5th JUSSC Conf. on Biomechanics. 54-55 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Sato M.: "Shear stress induced changes in endothelial cell stiffness measured by atomic force microscopy"Proc. Bioengineering Conference. BMD-42. 343-344 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Sato M.: "Local mechanical properties measured by atomic force microscopy for cultured bovine endothelial cells exposed to shear stress"J. Biomechanics. 33. 127-135 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Sato M.: "Mechanical stress and actin filament dynamics in cultured endothelial cell"Proc. 4th Int. Symp. on Tissue Eng. for Therapeutic Use. (印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] N. Kataoka: "Mechanical stimuli and changes in cytoskeletal structure of endothelial cells"J. Jap. Soc. Biorheol.. 12(4). 234-236 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] N. Kataoka: "The change of F-actin distribution and morphology of cultured bovine aortic endothelial cells in the early stage of fluid shear stress exposure"Trans. Jap. Soc. Mech. Eng. Ser. B. 64(622). 1801-1808 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] N. Kataoka: "Effects of the change of flow direction on the morphology of endothelial cells - Effect of the interval time -"Jap. J. Med. Elec. Biol. Eng.. 36(3). 302-308 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K. Takeda: "Morphological responses of endothelial cells to flow exposure using cultured aortic wall segments"Jap. J. Med. Elec. Biol. Eng.. 36(3). 293-301 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] N. Kataoka: "The effect of flow direction on the morphological responses of cultured bovine aortic endothelial cells"Med. Biol. Eng. Comput.. 36(1). 122-128 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] N. Kataoka: "The morphological responses of cultured bovine aortic endothelial cells to fluid-imposed shear stress under sparse and colony conditions"JSME Int. J. Ser. C. 41(1). 76-82 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] N. Kataoka: "How do flow directions affect the morphology of cultured endothelial cells? - In relation to the blood flow condition at atherosclerotic regions -"Proc. of The 10th International Conference on Mechanics in Medicine and Biology. 133-136 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] N. Kataoka: "Effects of cell - cell and cell - substrate contacts on the response of cultured endothelial cells to fluid shear stress"Proc of the 5th JUSSC Conf on Biomech. 54-55 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] M. Sato: "Shear stress induced changes in endothelial cell stiffness measured by atomic force microscopy"Proceedings of Bioengineering Conference. BED-Vol. 42. 343-344 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] M. Sato: "Local mechanical properties measured by atomic force microscopy for cultured bovine endothelial cells exposed to shear stress"J Biomechanics. 33(1). 127-135 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] M. Sato: "Mechanical stress and actin filament dynamics in cultured endothelial cell"Proceedings of 4th International Symposium on Tissue Engineering for Therapeutic Use. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Sato M.: "Shear stress induced changes in endothelial cell stiffness measured by atomic force microscopy"Proc.Bioengineering Conterence. BMD-42. 343-344 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Sato M.: "Local mechanical properties measured by atomic force microscopy for cultured bovine indothelial cells exposed to shear stress"J. Biomechanica. 33・1. 127-135 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Sato M.: "Mechanical stress and actin filament dynamics in cultured endothelial cell"Proc. 4th Int. Symp. On Tissue Eng. For Therapeutic Use. (印刷中).

    • Related Report
      1999 Annual Research Report
  • [Publications] 片岡則之: "力学的刺激と内皮細胞骨格構造の変化" 日本バイオレオロジー学会誌. 12-4. 234-246 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 片岡則之: "せん断応力負荷の初期過程における培養内皮細胞の形態およびF-アクチンフィラメントの変化" 日本機械学会論文集(B編). 64-622. 1801-1808 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 片岡則之: "流れの方向変化が培養内皮細胞の形態に及ぼす影響-流れの方向変化の時間間隔の影響-" 医用電子と生体工学. 36-3. 302-308 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 武田浩一郎: "培養動脈組織片を用いた内皮細胞の流れに対する形態的反応の検討" 医用電子と生体工学. 36-3. 293-301 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kataoka,N.: "The effect of flow direction on the morphological responses of cultured bovine aortic endothelial cells" Medical & Biological Engineering & Computing. 36-1. 122-128 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kataoka,N.: "The morphological responses of cultured bovine aortic endothelial cells to fluid-imposed shear stress under sparse and colony conditions" JSME International Journal Series C. 41-1. 76-82 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kataoka,N.: "How do flow directions affect the morphology of cultured endothelial cells?-In relation to the blood flow condition at atherosclerotic regions-" Proc.of The 10th International Conference on Mechanics in Medicine and Biology. 133-136 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kataoka,N.: "Effects of cell-cell and cell-substrate contacts on the response of cultured endothelial cells to fluid shear stress" Proc of the 5th JUSSC Conf on Biomech. 54-55 (1998)

    • Related Report
      1998 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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