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Salient segmentation is a critical step in biomedical image analysis, aiming to cut out regions that are most interesting to humans. Recently, supervised methods have achieved promising results in biomedical areas, but they depend on annotated training data sets, which requires labor and proficiency in related background knowledge. In contrast, unsupervised learning makes data-driven decisions by obtaining insights directly from the data themselves. In this paper, we propose a completely unsupervised self-aware network based on pre-training and attentional backpropagation for biomedical salient segmentation, named as PUB-SalNet. Firstly, we aggregate a new biomedical data set from several simulated Cellular Electron Cryo-Tomography (CECT) data sets featuring rich salient objects, different SNR settings, and various resolutions, which is called SalSeg-CECT. Based on the SalSeg-CECT data set, we then pre-train a model specially designed for biomedical tasks as a backbone module to initialize network parameters. Next, we present a U-SalNet network to learn to selectively attend to salient objects. It includes two types of attention modules to facilitate learning saliency through global contrast and local similarity. Lastly, we jointly refine the salient regions together with feature representations from U-SalNet, with the parameters updated by self-aware attentional backpropagation. We apply PUB-SalNet for analysis of 2D simulated and real images and achieve state-of-the-art performance on simulated biomedical data sets. Furthermore, our proposed PUB-SalNet can be easily extended to 3D images. The experimental results on the 2d and 3d data sets also demonstrate the generalization ability and robustness of our method. 

Abstract Background Cryo-electron tomography is an important and powerful technique to explore the structure, abundance, and location of ultrastructure in a near-native state. It contains detailed information of all macromolecular complexes in a sample cell. However, due to the compact and crowded status, the missing edge effect, and low signal to noise ratio (SNR), it is extremely challenging to recover such information with existing image processing methods. Cryo-electron tomogram simulation is an effective solution to test and optimize the performance of the above image processing methods. The simulated images could be regarded as the labeled data which covers a wide range of macromolecular complexes and ultrastructure. To approximate the crowded cellular environment, it is very important to pack these heterogeneous structures as tightly as possible. Besides, simulating non-deformable and deformable components under a unified framework also need to be achieved. Result In this paper, we proposed a unified framework for simulating crowded cryo-electron tomogram images including non-deformable macromolecular complexes and deformable ultrastructures. A macromolecule was approximated using multiple balls with fixed relative positions to reduce the vacuum volume. A ultrastructure, such as membrane and filament, was approximated using multiple balls with flexible relative positions so that this structure could deform under force field. In the experiment, 400 macromolecules of 20 representative types were packed into simulated cytoplasm by our framework, and numerical verification proved that our method has a smaller volume and higher compression ratio than the baseline single-ball model. We also packed filaments, membranes and macromolecules together, to obtain a simulated cryo-electron tomogram image with deformable structures. The simulated results are closer to the real Cryo-ET, making the analysis more difficult. The DOG particle picking method and the image segmentation method are tested on our simulation data, and the experimental results show that these methods still have much room for improvement. Conclusion The proposed multi-ball model can achieve more crowded packaging results and contains richer elements with different properties to obtain more realistic cryo-electron tomogram simulation. This enables users to simulate cryo-electron tomogram images with non-deformable macromolecular complexes and deformable ultrastructures under a unified framework. To illustrate the advantages of our framework in improving the compression ratio, we calculated the volume of simulated macromolecular under our multi-ball method and traditional single-ball method. We also performed the packing experiment of filaments and membranes to demonstrate the simulation ability of deformable structures. Our method can be used to do a benchmark by generating large labeled cryo-ET dataset and evaluating existing image processing methods. Since the content of the simulated cryo-ET is more complex and crowded compared with previous ones, it will pose a greater challenge to existing image processing methods.