2.06 Å Cryo-EM Structural Determination by a Conventional Scintillator-Coupled CMOS Detector

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2.06 Å Cryo-EM Structural Determination by a Conventional Scintillator-Coupled CMOS Detector

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Author(s): Shinji Aramaki ¹ ^, , Yuya Yoshida ² , Tomohito Tanihara ² , Kosuke Oyama ² , Kaita Otsuki ² , Yama Terada ² , Naoya Matsunaga ² , Shigehiro Ohdo ² , Kouta Mayanagi ²

Publication date (Electronic): 21 January 2025

Conference name: 13th Asia Pacific Microscopy Congress 2025 (APMC13)

Conference date: 2-7 Febuary 2025

Keywords: Cryo-EM, Single Particle Analysis

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Abstract

In the past decade, a resolution revolution has transformed the field of cryo-electron microscopy (cryo-EM), establishing it as a pivotal technology in structural biology. Key drivers of this revolution include the development of direct electron detectors (DEDs), motion blur correction, automated acquisition software like SerialEM and Leginon, and advancements in image processing tools such as RELION and cryoSPARC. DEDs offer higher quantum efficiency compared to earlier detectors, such as CCD cameras and analog film. Anti-motion blur technologies, made possible through CMOS advancements that enable short frame readout times and precise frame alignment, reduce blur caused by instability in microscope goniometers and/or movement of the vitrified ice layer which is induced by electron beam irradiation. Automated acquisition software captures more images, achieving higher resolution while avoiding unexpected beam damage associated with manual microscope operation. Furthermore, advanced reconstruction software can further enhance resolution by refining aberration parameters, per-particle motion, and defocus determination.

In recent years, the resolution record for single particle analysis has been pushed further through the implementation of lower energy spread illumination techniques, such as monochromators, cold field emission guns (cold-FEG), lower energy-spread beam sources, and reductions in inelastic scattering facilitated by post-column or in-column energy filters [ 1]. These correction and filtering instruments enhance the parameters that affect the envelope function of the contrast transfer function (CTF), thus improving high-resolution information. Conversely, in efforts to break the atomic-resolution record, attention has also turned toward making cryo-EM more widely accessible, with the aim of democratizing these techniques. Presently, high-resolution structural analysis using 200 kV electron microscopes with conventional side-entry cryo transfer holders has become practical [ 2], and the integration of field emission guns (FEG) and hybrid pixelated detectors in 100 kV microscope columns has yielded high-resolution structures [ 3].

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Publication date (Electronic): 21 January 2025

Electronic Location Identifier: e031

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[1 ]TVIPS GmbH, Gilching, Germany

[2 ]Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan

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*Corresponding author: Shinji.Aramaki@ 123456tvips.com

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Shinji Aramaki https://orcid.org/0009-0004-5164-4794

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DOI: 10.14293/APMC13-2025-0031

SO-VID: 1da51e22-0831-46ce-bf1b-182dbe9de81a

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Published under Creative Commons Attribution 4.0 International ( CC BY 4.0). Users are allowed to share (copy and redistribute the material in any medium or format) and adapt (remix, transform, and build upon the material for any purpose, even commercially), as long as the authors and the publisher are explicitly identified and properly acknowledged as the original source.

Conference name: 13th Asia Pacific Microscopy Congress 2025

Conference acronym: APMC13

Conference number: 13

Conference location: Brisbane, Australia

Conference date: 2-7 Febuary 2025

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References

S Yonekura et al., Community Chemistry 6, 96 (2023), https://doi.org/10.1038/s42004-023-00900-x
Y Kayama et al., Scientific Report 11, 8395 (2021), https://doi.org/10.1038/s41598-021-87183-1
G McMullan et al., PNAS 120, 49 (2023), https://doi.org/10.1073/pnas.2312905120

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[1] S Yonekura et al., Community Chemistry 6, 96 (2023), https://doi.org/10.1038/s42004-023-00900-x

[2] Y Kayama et al., Scientific Report 11, 8395 (2021), https://doi.org/10.1038/s41598-021-87183-1

[3] G McMullan et al., PNAS 120, 49 (2023), https://doi.org/10.1073/pnas.2312905120

2.06 Å Cryo-EM Structural Determination by a Conventional Scintillator-Coupled CMOS Detector

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