| Project/Area Number |
17K07384
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Cell biology
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| Research Institution | Kyoto University |
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Principal Investigator |
Kiuchi Tai 京都大学, 医学研究科, 准教授 (70443984)
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| Project Period (FY) |
2017-04-01 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2019: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2017: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
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| Keywords | 超解像顕微鏡法 / アクチン / 細胞骨格 / ３次元イメージング / がん浸潤 / アクチン骨格 / 細胞移動 |
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| Outline of Final Research Achievements |
Cancer cells rearrange the cytoskeletons and focal adhesions in appropriate 3D patterns through interactions with the surroundings for cancer invasion. To study the 3D rearrangement mechanism of the cytoskeletons, multitargets super-resolution imaging IRIS was developed into 3D imaging. For the 3D imaging, HILO illumination and adaptive optics were inserted into the optical path of the microscope. To reduce the background light, the Z resolution and Z drift, the excitation conditions were examined in detail. Then, the 3D networks of actin filaments and microtubules in the apical region of cells were visualized. Furthermore, it was found that EGF receptors, which induce the rearrangement of cytoskeletons, are accumulated in the interspace between actin stress fibers by EGF stimulation and then are endocytosed.
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| Academic Significance and Societal Importance of the Research Achievements |
これまで超解像顕微鏡法を３次元化するために様々な取り組みが行われてきた。しかし、蛍光退色や背景光、Z分解能、測定中のZドリフトなど励起条件や測定条件で検討する課題が多い。さらに多重染色超解像イメージングは、標的の標識率や複数の蛍光色素を用いることによる色収差や球面収差の問題が追加される。IRISは、結合解離プローブを用いることで、標識率の上限を突破し、さらに同一の蛍光色素を結合解離プローブに接合することで、この色収差や球面収差の問題を回避している。IRISを３次元イメージングへと発展させることで、今後、細胞や組織の３次元的な構造の詳細が明らかになることが期待できる。
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