Deep Learning Based Supervised Semantic Segmentation of Electron Cryo-Subtomograms

Cellular Electron Cryo-Tomography (CECT) is a powerful imaging technique for the 3D visualization of cellular structure and organization at submolecular resolution. It enables analyzing the native structures of macromolecular complexes and their spatial organization inside single cells. However, due to the high degree of structural complexity and practical imaging limitations, systematic macromolecular structural recovery inside CECT images remains challenging. Particularly, the recovery of a macromolecule is likely to be biased by its neighbor structures due to the high molecular crowding. To reduce the bias, here we introduce a novel 3D convolutional neural network inspired by Fully Convolutional Network and Encoder-Decoder Architecture for the supervised segmentation of macromolecules of interest in subtomograms. The tests of our models on realistically simulated CECT data demonstrate that our new approach has significantly improved segmentation performance compared to our baseline approach. Also, we demonstrate that the proposed model has generalization ability to segment new structures that do not exist in training data.

[1]  J. Frank,et al.  Signal-to-noise ratio of electron micrographs obtained by cross correlation , 1975, Nature.

[2]  T. Bhat,et al.  The Protein Data Bank and the challenge of structural genomics , 2000, Nature Structural Biology.

[3]  W. Baumeister,et al.  Prospects of electron cryotomography to visualize macromolecular complexes inside cellular compartments: implications of crowding. , 2002, Biophysical chemistry.

[4]  G Sapiro,et al.  Classification and 3D averaging with missing wedge correction in biological electron tomography. , 2008, Journal of structural biology.

[5]  R. Aebersold,et al.  Visual proteomics of the human pathogen Leptospira interrogans , 2009, Nature Methods.

[6]  M. Valle,et al.  Averaging of electron subtomograms and random conical tilt reconstructions through likelihood optimization. , 2009, Structure.

[7]  G. Jensen,et al.  Electron tomography of cells , 2011, Quarterly Reviews of Biophysics.

[8]  Frank Alber,et al.  High-throughput subtomogram alignment and classification by Fourier space constrained fast volumetric matching. , 2012, Journal of structural biology.

[9]  V. Lučić,et al.  Cryo-electron tomography: The challenge of doing structural biology in situ , 2013, The Journal of cell biology.

[10]  J. Briggs Structural biology in situ--the potential of subtomogram averaging. , 2013, Current opinion in structural biology.

[11]  Min Xu,et al.  Automated target segmentation and real space fast alignment methods for high-throughput classification and averaging of crowded cryo-electron subtomograms , 2013, Bioinform..

[12]  Yuxiang Chen,et al.  Autofocused 3D classification of cryoelectron subtomograms. , 2014, Structure.

[13]  Peijun Zhang,et al.  Correlative Fluorescence and Electron Microscopy , 2014, Current protocols in cytometry.

[14]  Trevor Darrell,et al.  Fully convolutional networks for semantic segmentation , 2015, 2015 IEEE Conference on Computer Vision and Pattern Recognition (CVPR).

[15]  Andrew Zisserman,et al.  Very Deep Convolutional Networks for Large-Scale Image Recognition , 2014, ICLR.

[16]  Y. Sugita,et al.  Biomolecular interactions modulate macromolecular structure and dynamics in atomistic model of a bacterial cytoplasm , 2016, eLife.

[17]  Yuan Yu,et al.  TensorFlow: A system for large-scale machine learning , 2016, OSDI.

[18]  Min Xu,et al.  Simulating cryo electron tomograms of crowded cell cytoplasm for assessment of automated particle picking , 2016, BMC Bioinformatics.

[19]  Min Xu,et al.  De Novo Structural Pattern Mining in Cellular Electron Cryotomograms. , 2015, Structure.

[20]  Roberto Cipolla,et al.  SegNet: A Deep Convolutional Encoder-Decoder Architecture for Image Segmentation , 2015, IEEE Transactions on Pattern Analysis and Machine Intelligence.

[21]  Wei Dai,et al.  Convolutional Neural Networks for Automated Annotation of Cellular Cryo-Electron Tomograms , 2017, Nature Methods.

[22]  Karim Elmaaroufi,et al.  Improved deep learning-based macromolecules structure classification from electron cryo-tomograms , 2017, Machine Vision and Applications.

[23]  Min Xu,et al.  A convolutional autoencoder approach for mining features in cellular electron cryo-tomograms and weakly supervised coarse segmentation , 2017, Journal of structural biology.

[24]  Min Xu,et al.  De Novo Structural Pattern Mining in Cellular Electron Cryotomograms. , 2019, Structure.