| Budget Amount *help |
¥18,070,000 (Direct Cost: ¥13,900,000、Indirect Cost: ¥4,170,000)
Fiscal Year 2014: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000)
Fiscal Year 2013: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2012: ¥6,760,000 (Direct Cost: ¥5,200,000、Indirect Cost: ¥1,560,000)
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| Outline of Final Research Achievements |
Actin dynamics contribute to many cellular processes, while the detail mechanism of actin polymerization is unknown. In this study, single molecule imaging of actin polymerization was performed to answer this question.
As critical concentration of actin is >100 nM, it is not possible to observe actin polymerization at single molecule level by total internal reflection fluorescence microscopy. Hence, we applied a linear zero-mode waveguide (ZMW), which has a 100-nm wide slit structure, allowing single molecule imaging at higher concentrations.
We found that single BODIPY-FL-labelled actin was observed in a linear ZMW at 100 nM. F-actin was immobilized in a linear ZMW. Actin elongated in a linear ZMW at a similar elongation rate on a cover glass. Actin polymerization at a single molecule level in a linear ZMW is now under investigation.
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