Removing Contamination-Induced Reconstruction Artifacts from Cryo-electron Tomograms

Imaging of fully hydrated, vitrified biological samples by electron tomography yields structural information about cellular protein complexes in situ. Here we present a computational procedure that removes artifacts of three-dimensional reconstruction caused by contamination present in samples durin...

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Bibliographic Details
Published in:Biophysical journal 2016-02, Vol.110 (4), p.850-859
Main Authors: Fernandez, Jose-Jesus, Laugks, Ulrike, Schaffer, Miroslava, Bäuerlein, Felix J.B., Khoshouei, Maryam, Baumeister, Wolfgang, Lucic, Vladan
Format: Article
Language:English
Subjects:
Animals
Cell Biophysics
Cells
Contamination
Electron Microscope Tomography - methods
Electron microscopes
Fiducial Markers
HeLa Cells
Humans
Image Processing, Computer-Assisted
Male
Neurons - cytology
Phantoms, Imaging
Proteins
Rats
Rats, Wistar
Tomography
Vitrification
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Description
Summary:Imaging of fully hydrated, vitrified biological samples by electron tomography yields structural information about cellular protein complexes in situ. Here we present a computational procedure that removes artifacts of three-dimensional reconstruction caused by contamination present in samples during imaging by electron microscopy. Applying the procedure to phantom data and electron tomograms of cellular samples significantly improved the resolution and the interpretability of tomograms. Artifacts caused by surface contamination associated with thinning by focused ion beam, as well as those arising from gold fiducial markers and from common, lower contrast contamination, could be removed. Our procedure is widely applicable and is especially suited for applications that strive to reach a higher resolution and involve the use of recently developed, state-of-the-art instrumentation.
ISSN:0006-3495
1542-0086
DOI:10.1016/j.bpj.2015.10.043