Benefits and Limitations of Low-kV Macromolecular Imaging of Frozen-Hydrated Biological Samples

Biophysical Journal
E MajorovitsR R Schröder

Abstract

Object contrast is one of the most important parameters of macromolecular imaging. Low-voltage transmission electron microscopy has shown an increased atom contrast for carbon materials, indicating that amplitude contrast contributions increase at a higher rate than phase contrast and inelastic scattering. Here, we studied image contrast using ice-embedded tobacco mosaic virus particles as test samples at 20-80 keV electron energy. The particles showed the expected increase in contrast for lower energies, but at the same time the 2.3-nm-resolution measure decayed more rapidly. We found a pronounced signal loss below 60 keV, and therefore we conclude that increased inelastic scattering counteracts increased amplitude contrast. This model also implies that as long as the amplitude contrast does not increase with resolution, beam damage and multiple scattering will always win over increased contrast at the lowest energies. Therefore, we cannot expect that low-energy imaging of conventionally prepared samples would provide better data than state-of-the-art 200-300 keV imaging.

References

Apr 27, 2000·Ultramicroscopy·I AngertR R Schröder
Sep 22, 2006·Journal of Structural Biology·Koji YonekuraDavid A Agard
Mar 28, 2009·Science·Caglar O GiritA Zettl
Apr 8, 2011·Physical Review Letters·J KotakoskiJ C Meyer
Feb 24, 2012·Ultramicroscopy·David C BellDmitry V Kolmykov
Aug 23, 2012·Ultramicroscopy·R F Egerton
May 11, 2013·Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada·Jean-Pierre BaudoinNiels de Jonge
Jan 3, 2014·Microscopy and Microanalysis : the Official Journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada·Nicole FrindtRasmus R Schröder
Jan 11, 2014·Science·David Scanlan
Oct 22, 2014·Proceedings of the National Academy of Sciences of the United States of America·Radostin DanevWolfgang Baumeister
Dec 22, 2014·Journal of Structural Biology·Simon A FrommCarsten Sachse

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Citations

Jun 28, 2017·Ultramicroscopy·J A MirA I Kirkland

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