Development for single-molecule deep imaging method of biological sample under cryogenic conditions
| Project/Area Number |
19K22162
|
|---|
| Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
|
| Allocation Type | Multi-year Fund |
|---|
| Review Section |
Medium-sized Section 32:Physical chemistry, functional solid state chemistry, and related fields
|
|---|
| Research Institution | Tokyo Institute of Technology |
|---|
Principal Investigator |
|
|---|
| Project Period (FY) |
2019-06-28 – 2021-03-31
|
| Project Status |
Completed (Fiscal Year 2020)
|
| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2020: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Fiscal Year 2019: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
|
|---|
| Keywords | １分子観察 / 可変浸レンズ / 蛍光顕微鏡 / 三次元イメージング / 生体試料 / 顕微鏡 |
|---|
| Outline of Research at the Start |
蛍光顕微鏡で観察する試料の表面には、空気/水（または油/水などの）界面が存在する。試料表面は平らな場合が多く、平らな界面で光が屈折すると収差が産まれ、顕微鏡の分解能と検出効率が大きく低下する このため、蛍光顕微鏡でも、細胞や組織のような厚みのある試料全体を正しく画像化することは難しい。そこで、当該研究では、界面屈折の影響をゼロにできるWILを開発する。
|
| Outline of Final Research Achievements |
Three-dimensional optical microscopy with a high numerical aperture (NA) remains challenging for thick biological specimens owing to aberrations arising from interface refractions. In this project, we have developed a variable immersion lens (VIL) for deep fluorescence imaging A VIL is a high-NA concentric meniscus lens and was used in combination with an aberration-corrected high-NA reflecting objective (TORA-FUJI mirror). VIL microscope enables diffraction-limited 1.2-NA imaging in water (refractive index of 1.34) at a depth of 0.3 mm by minimizing aberrations due to refraction of a sample interface. Another aberration due to refractive index mismatching between a mounting medium and an object can be also corrected by the VIL system because various fluids with different refractive indexes can be used as mounting media for the VIL. As a result, we have demonstrated that a cell spheroid can be imaged at a true dimension.
|
| Academic Significance and Societal Importance of the Research Achievements |
これまで細胞内の小器官を測定する場合、培養細胞ぐらいの厚さ（１０ミクロン）がせいぜいで組織レベルの試料になると細胞ごとの分解能が限界であった。我々の提案する方法は生体組織の中の小器官を鮮明に取得できる方法であり、とても大きな意義があると考えている。この技術は医学、薬学にもすぐに応用できる方法であり、こちらの方向を伸ばすことで社会に貢献したい。
|
Report
(2 results)
Research Products
(10 results)
