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Near-Atomic Resolution Structure Determination in Over-Focus with Volta Phase Plate by Cs-corrected Cryo-EM

Xiao Fan, Lingyun Zhao, Chuan Liu, Jin-Can Zhang, Kelong Fan, Xiyun Yan, Hai-Lin Peng, Jianlin Lei, Hong-Wei Wang
doi: https://doi.org/10.1101/148205
Xiao Fan
1Ministry of Education Key Laboratory of Protein Sciences, Tsinghua-Peking Joint Center for Life Sciences, Beijing Advanced Innovation Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China
2Tsinghua-Peking Joint Center for Life Sciences, Tsinghua University, Beijing 100084, China
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Lingyun Zhao
1Ministry of Education Key Laboratory of Protein Sciences, Tsinghua-Peking Joint Center for Life Sciences, Beijing Advanced Innovation Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China
2Tsinghua-Peking Joint Center for Life Sciences, Tsinghua University, Beijing 100084, China
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Chuan Liu
1Ministry of Education Key Laboratory of Protein Sciences, Tsinghua-Peking Joint Center for Life Sciences, Beijing Advanced Innovation Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China
2Tsinghua-Peking Joint Center for Life Sciences, Tsinghua University, Beijing 100084, China
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Jin-Can Zhang
3Center for Nanochemistry, Beijing Science and Engineering Center for Nanocarbons, Beijing National Laboratory for Molecular Sciences, College of Chemistry and Molecular Engineering, Peking University, Beijing, 100871, China.
4Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, 100871, China.
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Kelong Fan
5Key Laboratory of Protein and Peptide Pharmaceuticals, CAS-University of Tokyo Joint Laboratory of Structural Virology and Immunology, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China
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Xiyun Yan
5Key Laboratory of Protein and Peptide Pharmaceuticals, CAS-University of Tokyo Joint Laboratory of Structural Virology and Immunology, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China
6University of Chinese Academy of Sciences, Beijing 100049, China School of Chemistry and Joint Graduate Program of Peking-Tsinghua-National Institute of Biological Science, Tsinghua University, Beijing 100084, China
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Hai-Lin Peng
3Center for Nanochemistry, Beijing Science and Engineering Center for Nanocarbons, Beijing National Laboratory for Molecular Sciences, College of Chemistry and Molecular Engineering, Peking University, Beijing, 100871, China.
4Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, 100871, China.
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Jianlin Lei
1Ministry of Education Key Laboratory of Protein Sciences, Tsinghua-Peking Joint Center for Life Sciences, Beijing Advanced Innovation Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China
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  • For correspondence: jllei@tsinghua.edu.cn hongweiwang@tsinghua.edu.cn
Hong-Wei Wang
1Ministry of Education Key Laboratory of Protein Sciences, Tsinghua-Peking Joint Center for Life Sciences, Beijing Advanced Innovation Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China
2Tsinghua-Peking Joint Center for Life Sciences, Tsinghua University, Beijing 100084, China
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  • For correspondence: jllei@tsinghua.edu.cn hongweiwang@tsinghua.edu.cn
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Summary

Volta phase plate (VPP) is a recently developed transmission electron microscope (TEM) apparatus that can significantly enhance the image contrast of biological samples in cryo-electron microscopy (cryo-EM) therefore impose the possibility to solve structures of relatively small macromolecules at high resolution. In this work, we performed theoretical analysis and found that using phase plate on objective lens spherical aberration (Cs)-corrected TEM may gain some interesting optical properties, including the over-focus imaging of macromolecules. We subsequently evaluated the imaging strategy of frozen-hydrated apo-ferritin with VPP on a Cs-corrected TEM and obtained the structure of apo-ferritin at near atomic resolution from both under- and over-focused dataset, illustrating the feasibility and new potential of combining VPP with Cs-corrected TEM for high resolution cryo-EM.

Highlights The successful combination of volta phase plate and Cs-corrector in single particle cryo-EM.

Near-atomic structure determined from over-focused images by cryo-EM. VPP-Cs-corrector coupled EM provides interesting optical properties.

In Brief We took the unique advantage of the optical system by combining the volta phase plate and Cs-corrector in a modern TEM to collect high resolution micrographs of frozen-hydrated apo-ferritin in over-focus imaging conditions and determined the structure of apo-ferritin at 3.0 Angstrom resolution.

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Posted June 09, 2017.
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Near-Atomic Resolution Structure Determination in Over-Focus with Volta Phase Plate by Cs-corrected Cryo-EM
Xiao Fan, Lingyun Zhao, Chuan Liu, Jin-Can Zhang, Kelong Fan, Xiyun Yan, Hai-Lin Peng, Jianlin Lei, Hong-Wei Wang
bioRxiv 148205; doi: https://doi.org/10.1101/148205
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Near-Atomic Resolution Structure Determination in Over-Focus with Volta Phase Plate by Cs-corrected Cryo-EM
Xiao Fan, Lingyun Zhao, Chuan Liu, Jin-Can Zhang, Kelong Fan, Xiyun Yan, Hai-Lin Peng, Jianlin Lei, Hong-Wei Wang
bioRxiv 148205; doi: https://doi.org/10.1101/148205

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