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-Structure paper
タイトル | Accurate magnification determination for cryoEM using gold. |
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ジャーナル・号・ページ | Ultramicroscopy, Vol. 256, Page 113883, Year 2024 |
掲載日 | 2023年11月15日 |
著者 | Joshua L Dickerson / Erin Leahy / Mathew J Peet / Katerina Naydenova / Christopher J Russo / |
PubMed 要旨 | Determining the correct magnified pixel size of single-particle cryoEM micrographs is necessary to maximize resolution and enable accurate model building. Here we describe a simple and rapid ...Determining the correct magnified pixel size of single-particle cryoEM micrographs is necessary to maximize resolution and enable accurate model building. Here we describe a simple and rapid procedure for determining the absolute magnification in an electron cryomicroscope to a precision of <0.5%. We show how to use the atomic lattice spacings of crystals of thin and readily available test specimens, such as gold, as an absolute reference to determine magnification for both room temperature and cryogenic imaging. We compare this method to other commonly used methods, and show that it provides comparable accuracy in spite of its simplicity. This magnification calibration method provides a definitive reference quantity for data analysis and processing, simplifies the combination of multiple datasets from different microscopes and detectors, and improves the accuracy with which the contrast transfer function of the microscope can be determined. We also provide an open source program, magCalEM, which can be used to accurately estimate the magnified pixel size of a cryoEM dataset ex post facto. |
リンク | Ultramicroscopy / PubMed:38008055 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 1.5 - 1.8 Å |
構造データ | EMDB-17992: Structure of K27A mutant E.coli DPS EMDB-17995: Cryo-EM structure of mouse heavy-chain apoferritin |
由来 |
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