Hierarchical detection and analysis of macromolecular complexes in cryo-electron tomograms using Pyto software

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 co...

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Bibliographic Details
Published in:Journal of structural biology 2016-12, Vol.196 (3), p.503-514
Main Authors: Lučić, Vladan, Fernández-Busnadiego, Rubén, Laugks, Ulrike, Baumeister, Wolfgang
Format: Article
Language:English
Subjects:
Algorithms
Cryo
Cryoelectron Microscopy - methods
Electron Microscope Tomography - methods
Electron tomography
Image processing
Image Processing, Computer-Assisted - methods
Imaging, Three-Dimensional - methods
Macromolecular Substances - chemistry
Macromolecular Substances - isolation & purification
Segmentation
Software
Synapses
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Summary: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.
ISSN:1047-8477
1095-8657
DOI:10.1016/j.jsb.2016.10.004