Abstract Motivation Cryo-electron tomography (Cryo-ET) with sub-tomogram averaging (STA) is indispensable when studying macromolecule structures and functions in their native environments. However, current tomographic reconstructions suffer the low signal-to-noise (SNR) ratio and the missing wedge artifacts. Hence, automatic and accurate macromolecule localization and classification become the bottleneck problem for structural determination by STA. Here, we propose a 3D multi-scale dense convolutional neural network (MSDNet) for voxel-wise annotations of tomograms. Weighted focal loss is adopted as a loss function to solve the class imbalance. The proposed network combines 3D hybrid dilated convolutions (HDC) and dense connectivity to ensure an accurate performance with relatively few trainable parameters. 3D HDC expands the receptive field without losing resolution or learning extra parameters. Dense connectivity facilitates the re-use of feature maps to generate fewer intermediate feature maps and trainable parameters. Then, we design a 3D MSDNet based approach for fully automatic macromolecule localization and classification, called VP-Detector (Voxel-wise Particle Detector). VP-Detector is efficient because classification performs on the pre-calculated coordinates instead of a sliding window. Results We evaluated the VP-Detector on simulated tomograms. Compared to the state-of-the-art methods, our method achieved a competitive performance on localization with the highest F1-score. We also demonstrated that the weighted focal loss improves the classification of hard classes. We trained the network on a part of training sets to prove the availability of training on relatively small datasets. Moreover, the experiment shows that VP-Detector has a fast particle detection speed, which costs less than 14 minutes on a test tomogram. Contact zsh@amss.ac.cn , xfcui@email.sdu.edu.cn , zhangfa@ict.ac.cn Supplementary information Supplementary data are available at Bioinformatics online.

VP-Detector: A 3D convolutional neural network for automated macromolecule localization and classification in cryo-electron tomograms