2001 Fiscal Year Final Research Report Summary
Development of a method to measure viscoelasticity of cardiomyocytes using scanning probe microscope
| Project/Area Number |
12557066
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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 | 展開研究 |
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| Research Field |
Circulatory organs internal medicine
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| Research Institution | National Cardiovascular Center Research Institute |
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Principal Investigator |
SUGIMACHI Masaru National Cardiovascular Center Research Institute, Cardiovascular Dynamics, Senior Staff, 循環動態機能部, 室長 (40250261)
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| Co-Investigator(Kenkyū-buntansha) |
SHISHIDO Toshiaki National Cardiovascular Center Research Institute, Cardiovascular Dynamics, Staff, 循環動態機能部, 室員 (60300977)
SUNAGAWA Kenji National Cardiovascular Center Research Institute, Cardiovascular Dynamics, Director, 循環動態機能部, 部長 (50163043)
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| Project Period (FY) |
2000 – 2001
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| Keywords | scanning probe microscope / minute vibration / time-varying elastance / viscosity / cardiomyocyte / laminin / collagenase / biopsy specimen |
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| Research Abstract |
We developed a method to measure viscoelasticity of cardiomyocytes from animals. Rats were anesthetized with pentobarbital, and the hearts were retrogradely perfused with collagenase to isolate cells. The cells were incubated for 4 hour (37℃, 5 %CC_2) in a laminin-coated dish, during which the cells firmly attached to the dish. We made a circuit and modified a scanning probe microscope so that we were able to control the vertical position of the probe with a piezoelectric device according to external command. We also developed a software to impose command and to measure the position of the piezoelectric device and the bending of the cantilever. When we imposed a sinusoidal vibration, the bending of the cantilever was in phase, indicating the viscosity of the superfusate is negligible.
We measured the changes in elasticity of cells during cardiac cycle by subjecting the cell to electrical stimulation, but the results indicated no significant changes. As it is known osmotic stress do change elasticity, we conjectured that the probe detects the local elasticity, and the changes in probe size seem necessary to detect the myocardial cellular elasticity changes. The lack of preload seems to contribute the small changes in elasticity.
We were not able to collect sufficient healthy cells either by superfusion with collagenase with or without ultrasonic agitation, or super fusion with protease.
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