Joshua L. Dickerson, Erin Leahy, Mathew J. Peet, Katerina Naydenova, Christopher J. Russo
{"title":"Accurate magnification determination for cryoEM using gold","authors":"Joshua L. Dickerson, Erin Leahy, Mathew J. Peet, Katerina Naydenova, Christopher J. Russo","doi":"10.1016/j.ultramic.2023.113883","DOIUrl":null,"url":null,"abstract":"<div><p>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 <span><math><mo><</mo></math></span>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.</p></div>","PeriodicalId":23439,"journal":{"name":"Ultramicroscopy","volume":null,"pages":null},"PeriodicalIF":2.1000,"publicationDate":"2023-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0304399123002000/pdfft?md5=f6ed5925a61508100520bd4110d6ad75&pid=1-s2.0-S0304399123002000-main.pdf","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Ultramicroscopy","FirstCategoryId":"5","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0304399123002000","RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MICROSCOPY","Score":null,"Total":0}
引用次数: 0
Abstract
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.
期刊介绍:
Ultramicroscopy is an established journal that provides a forum for the publication of original research papers, invited reviews and rapid communications. The scope of Ultramicroscopy is to describe advances in instrumentation, methods and theory related to all modes of microscopical imaging, diffraction and spectroscopy in the life and physical sciences.