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Ultramicroscopy. 2019 Aug;203:125-131. doi: 10.1016/j.ultramic.2019.02.007. Epub 2019 Feb 7.

The energy dependence of contrast and damage in electron cryomicroscopy of biological molecules.

Author information

1
MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.
2
MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK. Electronic address: crusso@mrc-lmb.cam.ac.uk.

Abstract

We have measured the dependence on electron energy of elastic and inelastic scattering cross-sections from carbon, over the energy range that includes 100 keV to 300 keV. We also compared quantitatively the radiation damage to bacteriorhodopsin and paraffin (C44H90) at 100 keV and 300 keV by observing the fading of the diffraction spots from two-dimensional crystals as a function of electron fluence. The elastic cross-section is 2.01 - fold greater at 100 keV than at 300 keV, whereas the radiation damage increased by only 1.57. This implies that the amount of useful information from diffraction patterns or images of most biological structures should be 25% greater using 100 keV rather than 300 keV electrons. Using these measurements, we calculate the energy dependence of the available information per unit damage for a specimen of a particular thickness, which we call the "information coefficient." This allows us to determine the optimal energy for imaging a biological specimen of a given thickness. We find that for most single particle cryoEM specimens, 100 keV provides not only the highest potential for information per unit damage, but would also simplify the instrument while retaining the potential to reach high resolution with a minimum of data. These measurements will help guide the development and use of electron cryomicroscopes for biology.

KEYWORDS:

Electron scattering cross-section; Low-dose electron microscopy; Radiation damage; Structure determination; cryoEM

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