title: Hierarchical detection and analysis of macromolecular complexes in cryo-electron tomograms using Pyto software.
abstract: Molecular complexes, arguably the basic units carrying cellular function, can be visualized directly in their native environment by cryo-electron tomography. Here we describe a procedure for the detection of small, pleomorphic membrane-bound molecular complexes in cryo-tomograms by a hierarchical connectivity segmentation. Validation on phantom and real data showed above 90% true positive rates. This segmentation procedure is implemented in the Pyto software package, together with methods for quantitative characterization and classification of complexes detected by our segmentation procedure and for statistical analysis between experimental conditions. Therefore, the methods presented provide a means for the detection and quantitative interpretation of structures captured in cryo-electron tomograms, as well as for the elucidation of their cellular function.
authors.affiliation: Max-Planck-Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany. Electronic address: vladan@biochem.mpg.de.
authors.forename: Vladan
authors.initials: V
authors.lastname: Lučić
authors.affiliation: Max-Planck-Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
authors.forename: Rubén
authors.initials: R
authors.lastname: Fernández-Busnadiego
authors.affiliation: Max-Planck-Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Laugks
authors.affiliation: Max-Planck-Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Baumeister
mesh_terms: D000465:Algorithms; D020285:Cryoelectron Microscopy / Q000379:methods*; D055032:Electron Microscope Tomography / Q000379:methods*; D007091:Image Processing, Computer-Assisted / Q000379:methods; D021621:Imaging, Three-Dimensional / Q000379:methods; D046911:Macromolecular Substances / Q000737:chemistry* / Q000302:isolation & purification; D012984:Software
chemical_list: D046911:Macromolecular Substances
keywords: Cryo; Electron tomography; Image processing; Segmentation; Synapses