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J Struct Biol. 2017 Feb;197(2):135-144. doi: 10.1016/j.jsb.2016.06.005. Epub 2016 Jun 8.

Dynamo Catalogue: Geometrical tools and data management for particle picking in subtomogram averaging of cryo-electron tomograms.

Author information

1
BioEM Lab at C-CINA, Biozentrum, University of Basel, Mattenstrasse 26, CH-4058 Basel, Switzerland; Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, Mattenstrasse 26, CH-4058 Basel, Switzerland. Electronic address: daniel.castano@unibas.ch.
2
Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, Mattenstrasse 26, CH-4058 Basel, Switzerland; Max Planck Institute for Biophysics, Max-von-Laue-Straße 3, 60438 Frankfurt am Main, Germany; Buchman Institute for Molecular Life Sciences, Goethe University of Frankfurt, Max-von-Laue-Straße 15, 60438 Frankfurt am Main, Germany.
3
Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, Mattenstrasse 26, CH-4058 Basel, Switzerland.

Abstract

Cryo electron tomography allows macromolecular complexes within vitrified, intact, thin cells or sections thereof to be visualized, and structural analysis to be performed in situ by averaging over multiple copies of the same molecules. Image processing for subtomogram averaging is specific and cumbersome, due to the large amount of data and its three dimensional nature and anisotropic resolution. Here, we streamline data processing for subtomogram averaging by introducing an archiving system, Dynamo Catalogue. This system manages tomographic data from multiple tomograms and allows visual feedback during all processing steps, including particle picking, extraction, alignment and classification. The file structure of a processing project file structure includes logfiles of performed operations, and can be backed up and shared between users. Command line commands, database queries and a set of GUIs give the user versatile control over the process. Here, we introduce a set of geometric tools that streamline particle picking from simple (filaments, spheres, tubes, vesicles) and complex geometries (arbitrary 2D surfaces, rare instances on proteins with geometric restrictions, and 2D and 3D crystals). Advanced functionality, such as manual alignment and subboxing, is useful when initial templates are generated for alignment and for project customization. Dynamo Catalogue is part of the open source package Dynamo and includes tools to ensure format compatibility with the subtomogram averaging functionalities of other packages, such as Jsubtomo, PyTom, PEET, EMAN2, XMIPP and Relion.

KEYWORDS:

Cryo electron tomography; Data management; Subtomogram averaging

PMID:
27288866
DOI:
10.1016/j.jsb.2016.06.005
[Indexed for MEDLINE]

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