Tilt-series and electron microscope alignment for the correction of the non-perpendicularity of beam and tilt-axis

Journal of Structural Biology
Daniel Castaño DíezAchilleas S Frangakis

Abstract

In electron tomography the sample is tilted in the electron microscope and projections are recorded at different viewing angles. In the correct geometric setting, the tilt-axis of the object under scrutiny is perpendicular to the beam direction. However, we will demonstrate that this does not necessarily apply to all electron microscopes equipped with the default column alignment. The resulting effect is that a conical tilt is performed, which has to be considered in the reconstruction to avoid artifacts and to improve the resolution. A novel solution, with significantly improved convergence properties, will be introduced for calculating the three-dimensional marker model, which is necessary for the alignment of the tilt-series. Thereby, the angle between the beam direction and the tilt-axis is calculated, together with other geometrical distortions, like magnification and rotation changes, and incorporated in the reconstruction. Hereby, artifacts can be eliminated at the image processing basis, and the resolution can be significantly improved at the medium to high range frequencies. Synthetical and real data are used to demonstrate the obstructions caused by this effect and the quality improvement of the reconstructions. Final...Continue Reading

References

Oct 1, 1995·Ultramicroscopy·P PenczekJ Frank
Nov 23, 2000·Proceedings of the National Academy of Sciences of the United States of America·J BohmW Baumeister
Feb 28, 2001·Proceedings of the National Academy of Sciences of the United States of America·B J MarshJ R McIntosh
Dec 5, 2002·Current Opinion in Structural Biology·Wolfgang Baumeister
Jun 15, 2004·Journal of Structural Biology·Qingxiong S ZhengDavid A Agard

❮ Previous
Next ❯

Citations

Feb 26, 2013·Journal of Structural Biology·Hanspeter Winkler, Kenneth A Taylor
Jun 5, 2012·Micron : the International Research and Review Journal for Microscopy·Jose-Jesus Fernandez
Mar 18, 2008·Journal of Structural Biology·Daniel Castaño-DíezAchilleas S Frangakis
Sep 18, 2007·Journal of Structural Biology·J Bernard HeymannAlasdair C Steven
Jul 27, 2007·Journal of Structural Biology·Daniel Castaño-DíezAchilleas S Frangakis
Jun 30, 2007·Journal of Structural Biology·A E YakushevskaA J Koster
Apr 3, 2007·Current Opinion in Structural Biology·Grant J Jensen, Ariane Briegel
Oct 5, 2014·Journal of Structural Biology·Michael Kunz, Achilleas S Frangakis
Dec 14, 2011·Journal of Structural Biology·Dilworth Y ParkinsonMark A Le Gros
Oct 25, 2016·Micron : the International Research and Review Journal for Microscopy·Misa Hayashida, Marek Malac
Nov 4, 2011·The Review of Scientific Instruments·Misa HayashidaToshiyuki Fujimoto
Mar 1, 2017·Proceedings of the National Academy of Sciences of the United States of America·Mingyu GuAdam Frost
Oct 18, 2020·Proceedings of the National Academy of Sciences of the United States of America·Mateusz SikoraAchilleas S Frangakis

❮ Previous
Next ❯

Related Concepts

Related Feeds

Antifungals (ASM)

An antifungal, also known as an antimycotic medication, is a pharmaceutical fungicide or fungistatic used to treat and prevent mycosis such as athlete's foot, ringworm, candidiasis, cryptococcal meningitis, and others. Discover the latest research on antifungals here.

Antifungals

An antifungal, also known as an antimycotic medication, is a pharmaceutical fungicide or fungistatic used to treat and prevent mycosis such as athlete's foot, ringworm, candidiasis, cryptococcal meningitis, and others. Discover the latest research on antifungals here.