title: Automated tracing of filaments in 3D electron tomography reconstructions using Sculptor and Situs.
abstract: The molecular graphics program Sculptor and the command-line suite Situs are software packages for the integration of biophysical data across spatial resolution scales. Herein, we provide an overview of recently developed tools relevant to cryo-electron tomography (cryo-ET), with an emphasis on functionality supported by Situs 2.7.1 and Sculptor 2.1.1. We describe a work flow for automatically segmenting filaments in cryo-ET maps including denoising, local normalization, feature detection, and tracing. Tomograms of cellular actin networks exhibit both cross-linked and bundled filament densities. Such filamentous regions in cryo-ET data sets can then be segmented using a stochastic template-based search, VolTrac. The approach combines a genetic algorithm and a bidirectional expansion with a tabu search strategy to localize and characterize filamentous regions. The automated filament segmentation by VolTrac compares well to a manual one performed by expert users, and it allows an efficient and reproducible analysis of large data sets. The software is free, open source, and can be used on Linux, Macintosh or Windows computers.
authors.affiliation: School of Biomedical Informatics, The University of Texas Health Science Center at Houston, 7000 Fannin St., Houston, TX 77030, USA.
authors.forename: Mirabela
authors.initials: M
authors.lastname: Rusu
authors.forename: Zbigniew
authors.initials: Z
authors.lastname: Starosolski
authors.forename: Manuel
authors.initials: M
authors.lastname: Wahle
authors.forename: Alexander
authors.initials: A
authors.lastname: Rigort
authors.forename: Willy
authors.initials: W
authors.lastname: Wriggers
mesh_terms: D000465:Algorithms; D020285:Cryoelectron Microscopy / Q000379:methods*; D003599:Cytoskeleton / Q000502:physiology*; D055032:Electron Microscope Tomography / Q000379:methods*; D008958:Models, Molecular; D012984:Software