title: Removing Contamination-Induced Reconstruction Artifacts from Cryo-electron Tomograms.
abstract: 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.
authors.affiliation: Centro Nacional de Biotecnologia (Consejo Superior de Investigaciones Científicas), Madrid, Spain. Electronic address: jj.fernandez@csic.es.
authors.forename: Jose-Jesus
authors.initials: JJ
authors.lastname: Fernandez
authors.affiliation: Max-Planck-Institute of Biochemistry, Martinsried, Germany.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Laugks
authors.affiliation: Max-Planck-Institute of Biochemistry, Martinsried, Germany.
authors.forename: Miroslava
authors.initials: M
authors.lastname: Schaffer
authors.affiliation: Max-Planck-Institute of Biochemistry, Martinsried, Germany.
authors.forename: Felix J B
authors.initials: FJ
authors.lastname: Bäuerlein
authors.affiliation: Max-Planck-Institute of Biochemistry, Martinsried, Germany.
authors.forename: Maryam
authors.initials: M
authors.lastname: Khoshouei
authors.affiliation: Max-Planck-Institute of Biochemistry, Martinsried, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Baumeister
authors.affiliation: Max-Planck-Institute of Biochemistry, Martinsried, Germany. Electronic address: vladan@biochem.mpg.de.
authors.forename: Vladan
authors.initials: V
authors.lastname: Lucic
mesh_terms: D000818:Animals; D016477:Artifacts; D055032:Electron Microscope Tomography / Q000379:methods*; D057918:Fiducial Markers; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008297:Male; D009474:Neurons / Q000166:cytology; D019047:Phantoms, Imaging; D051381:Rats; D017208:Rats, Wistar; D058989:Vitrification