2019 Fiscal Year Annual Research Report
Adaptive optics approach for high-throughput, high-resolution cryo-electron microscopy of biological macromolecules and complexes
| Project/Area Number |
19K21176
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| Allocation Type | Multi-year Fund |
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| Research Institution | The University of Tokyo |
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Principal Investigator |
Danev Radostin 東京大学, 大学院医学系研究科(医学部), 教授 (50415931)
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| Project Period (FY) |
2019-04-01 – 2020-03-31
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| Keywords | cryo-electron microscopy / molecular structure |
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| Outline of Annual Research Achievements |
The goal of this research was to improve the efficiency of electron cryo-microscopy (cryo-EM) experiments. Cryo-EM has become a fundamental method for high-resolution structure determination of biological macromolecules. However, to produce a high-quality result it requires acquisition of thousands of images from the sample. With previous methods this took several days. Cryo-EMs are expensive instruments. Reducing the time for data acquisition can speed up research and allow efficient use of research budgets. Previously, the typical throughput was ~1000 images/day. We developed and implemented a vastly improved “adaptive optics” method for automated image acquisition which increased the experimental throughput fivefold. The method uses optical shifting of the acquisition area, instead of the traditional mechanical stage movement, which saves significant amount of time. After development and testing of the new approach, we installed it on the main electron microscope of the new electron microscopy facility at the University of Tokyo. Our method became the standard automated data acquisition technique at the facility. We shared information about best practices and trained all experienced microscope users on its proper use. The data acquisition throughput increased to ~5500 images/day which enabled the acquisition of a complete dataset in one day. Consequently, the use efficiency of the facility was greatly improved, and this helped us and many other researchers to achieve outstanding high-resolution results in their molecular structural projects.
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