title: Automated correlation of single particle tilt pairs for Random Conical Tilt and Orthogonal Tilt Reconstructions.
abstract: One of the major methodological challenges in single particle electron microscopy is obtaining initial reconstructions which represent the structural heterogeneity of the dataset. Random Conical Tilt and Orthogonal Tilt Reconstruction techniques in combination with 3D alignment and classification can be used to obtain initial low-resolution reconstructions which represent the full range of structural heterogeneity of the dataset. In order to achieve statistical significance, however, a large number of 3D reconstructions, and, in turn, a large number of tilted image pairs are required. The extraction of single particle tilted image pairs from micrographs can be tedious and time-consuming, as it requires intensive user input even for semi-automated approaches. To overcome the bottleneck of manual selection of a large number of tilt pairs, we developed an algorithm for the correlation of single particle images from tilted image pairs in a fully automated and user-independent manner. The algorithm reliably correlates correct pairs even from noisy micrographs. We further demonstrate the applicability of the algorithm by using it to obtain initial references both from negative stain and unstained cryo datasets.
authors.affiliation: Max-Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Hauer
authors.forename: Christoph
authors.initials: C
authors.lastname: Gerle
authors.forename: Jan-Martin
authors.initials: JM
authors.lastname: Kirves
authors.forename: Holger
authors.initials: H
authors.lastname: Stark
mesh_terms: D000465:Algorithms; D003198:Computer Simulation; D020285:Cryoelectron Microscopy / Q000379:methods*; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D046529:Microscopy, Electron, Transmission / Q000379:methods*; D008958:Models, Molecular; D059629:Signal-To-Noise Ratio; D016963:Thermus thermophilus / Q000201:enzymology; D025262:Vacuolar Proton-Translocating ATPases / Q000648:ultrastructure
chemical_list: D025262:Vacuolar Proton-Translocating ATPases