Relative merits and limiting factors for x-ray and electron microscopy of thick, hydrated organic materials

Ultramicroscopy
Ming Du, C Jacobsen

Abstract

Electron and x-ray microscopes allow one to image the entire, unlabeled structure of hydrated materials at a resolution well beyond what visible light microscopes can achieve. However, both approaches involve ionizing radiation, so that radiation damage must be considered as one of the limits to imaging. Drawing upon earlier work, we describe here a unified approach to estimating the image contrast (and thus the required exposure and corresponding radiation dose) in both x-ray and electron microscopy. This approach accounts for factors such as plural and inelastic scattering, and (in electron microscopy) the use of energy filters to obtain so-called "zero loss" images. As expected, it shows that electron microscopy offers lower dose for specimens thinner than about 1 µm (such as for studies of macromolecules, viruses, bacteria and archaebacteria, and thin sectioned material), while x-ray microscopy offers superior characteristics for imaging thicker specimen such as whole eukaryotic cells, thick-sectioned tissues, and organs. The required radiation dose scales strongly as a function of the desired spatial resolution, allowing one to understand the limits of live and frozen hydrated specimen imaging. Finally, we consider the fac...Continue Reading

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Citations

Nov 7, 2019·Acta Crystallographica. Section D, Structural Biology·Maxim PolikarpovThomas R Schneider
Sep 16, 2020·Nature Neuroscience·Aaron T KuanWei-Chung Allen Lee
Jun 15, 2019·Nature Communications·Michal OdstrcilManuel Guizar-Sicairos
May 5, 2021·Acta Crystallographica. Section A, Foundations and Advances·John R Helliwell
May 7, 2021·Journal of Applied Crystallography·Ming DuChris Jacobsen
Sep 9, 2021·Journal of Cell Science·Valerie PanneelsGebhard Schertler
Jul 3, 2021·Journal of Synchrotron Radiation·Ming DuChris Jacobsen

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