In this work, we propose a disentangled latent optimization-based method for parameterizing grouped deforming 3D objects into shape and deformation factors in an unsupervised manner. Our approach involves the joint optimization of a generator network along with the shape and deformation factors, supported by specific regularization techniques. For efficient amortized inference of disentangled shape and deformation codes, we train two order-invariant PointNet-based encoder networks in the second stage of our method. We demonstrate several significant downstream applications of our method, including unsupervised deformation transfer, deformation classification, and explainability analyses. Extensive experiments conducted on 3D human, animal, and facial expression datasets demonstrate that our simple approach is highly effective in these downstream tasks, comparable or superior to existing methods with much higher complexity.

In recent years, the interpretability of Deep Neural Networks (DNNs) has garnered significant attention, particularly due to their widespread deployment in critical domains like healthcare, finance, and autonomous systems. To address the challenge of understanding how DNNs make decisions, Explainable AI (XAI) methods, such as saliency maps, have been developed to provide insights into the inner workings of these models. This paper introduces DiffCAM, a novel XAI method designed to overcome limitations in existing Class Activation Map (CAM)-based techniques, which often rely on decision boundary gradients to estimate feature importance. DiffCAM differentiates itself by considering the actual data distribution of the reference class, identifying feature importance based on how a target example differs from reference examples. This approach captures the most discriminative features without relying on decision boundaries or prediction results, making DiffCAM applicable to a broader range of models, including foundation models. Through extensive experiments, we demonstrate the superior performance and flexibility of DiffCAM in providing meaningful explanations across diverse datasets and scenarios.

, a novel contrastive learning-based method that implicitly parameterizes and disentangles both transformation and content. DualContrast achieves this by generating positive and negative pairs for content and transformation in both data and latent spaces. We demonstrate that existing self-supervised approaches fail under similar implicit parameterization, underscoring the necessity of our method. Through extensive experiments on 3D microscopic images of protein mixtures and additional shape-focused datasets beyond microscopy, we validate our claims and demonstrate the first in-principle fully unsupervised identification of different protein compositions and conformations in 3D microscopic images.

Cryo-electron tomography (cryo-ET) is confronted with the intricate task of unveiling novel structures. General class discovery (GCD) seeks to identify new classes by learning a model that can pseudo-label unannotated (novel) instances solely using supervision from labeled (base) classes. While 2D GCD for image data has made strides, its 3D counterpart remains unexplored. Traditional methods encounter challenges due to model bias and limited feature transferability when clustering unlabeled 2D images into known and potentially novel categories based on labeled data. To address this limitation and extend GCD to 3D structures, we propose an innovative approach that harnesses a pretrained 2D transformer, enriched by an effective weight inflation strategy tailored for 3D adaptation, followed by a decoupled prototypical network. Incorporating the power of pretrained weight-inflated Transformers, we further integrate CLIP, a vision-language model to incorporate textual information. Our method synergizes a graph convolutional network with CLIP's frozen text encoder, preserving class neighborhood structure. In order to effectively represent unlabeled samples, we devise semantic distance distributions, by formulating a bipartite matching problem for category prototypes using a decoupled prototypical network. Empirical results unequivocally highlight our method's potential in unveiling hitherto unknown structures in cryo-ET. By bridging the gap between 2D GCD and the distinctive challenges of 3D cryo-ET data, our approach paves novel avenues for exploration and discovery in this domain.

Localizing macromolecules in crowded cellular cryo-electron tomography (cryo-ET) images or tomograms is crucial for determining their in situ structures. Traditional template matching-based approaches for this task suffer from template-specific biases and have low throughput. Given these problems, learning-based solutions are necessary. However, the paucity of annotated data for training poses substantial challenges for such learning-based methods. Moreover, preparing extensively annotated cellular tomograms for training macromolecule localization methods is extremely time-consuming and burdensome due to the large volume and low signal-to-noise ratio of the tomograms. In this work, we developed TomoPicker, an annotation-efficient macromolecule localization method for tomograms. To achieve such annotation-efficiency, TomoPicker regards macromolecule localization as a voxel classification problem and solves it with two different positive-unlabeled learning approaches. We evaluated TomoPicker on two experimental cryo-ET datasets of crowded eukaryotic cells and one experimental dataset of relatively less crowded prokaryotic cell. We observed that, with only 10 annotated macromolecule locations, TomoPicker with positive unlabeled learning achieved a performance comparable to that of state-of-the-art supervised methods trained with several hundred annotations. In other words, TomoPicker achieved plausible segmentation with up to 98% less data compared with supervised learning-based methods. Furthermore, it demonstrated substantial improvements over existing learning-based macromolecule localization methods under sparse annotation scenarios.

In many real-life image analysis applications, particularly in biomedical research domains, the objects of interest undergo multiple transformations that alters their visual properties while keeping the semantic content unchanged. Disentangling images into semantic content factors and transformations can provide significant benefits into many domain-specific image analysis tasks. To this end, we propose a generic unsupervised framework, Harmony, that simultaneously and explicitly disentangles semantic content from multiple parameterized transformations. Harmony leverages a simple cross-contrastive learning framework with multiple explicitly parameterized latent representations to disentangle content from transformations. To demonstrate the efficacy of Harmony, we apply it to disentangle image semantic content from several parameterized transformations (rotation, translation, scaling, and contrast). Harmony achieves significantly improved disentanglement over the baseline models on several image datasets of diverse domains. With such disentanglement, Harmony is demonstrated to incentivize bioimage analysis research by modeling structural heterogeneity of macromolecules from cryo-ET images and learning transformation-invariant representations of protein particles from single-particle cryo-EM images. Harmony also performs very well in disentangling content from 3D transformations and can perform coarse and fast alignment of 3D cryo-ET subtomograms. Therefore, Harmony is generalizable to many other imaging domains and can potentially be extended to domains beyond imaging as well.

Cryo-Electron Tomography (cryo-ET) is an emerging 3D imaging technique which shows great potentials in structural biology research. One of the main challenges is to perform classification of macromolecules captured by cryo-ET. Re-cent efforts exploit deep learning to address this challenge. However, training reliable deep models usually requires a huge amount of labeled data in supervised fashion. Annotating cryo-ET data is arguably very expensive. Deep Active Learning (DAL) can be used to reduce labeling cost while not sacrificing the task performance too much. Nevertheless, most existing methods resort to auxiliary models or complex fashions (e.g. adversarial learning) for uncertainty estimation, the core of DAL. These models need to be highly customized for cryo-ET tasks which require 3D networks, and extra efforts are also indispensable for tuning these models, rendering a difficulty of deployment on cryo-ET tasks. To address these challenges, we propose a novel metric for data selection in DAL, which can also be leveraged as a regularizer of the empirical loss, further boosting the task model. We demonstrate the superiority of our method via extensive experiments on both simulated and real cryo-ET datasets. Our source Code and Appendix can be found at this URL.

3D subtomogram image alignment, clustering, and segmentation are vital to macromolecular structure recognition in cryo-electron tomography (cryo-ET). However, acquiring ground-truth labels to train a unified deep learning model that can simultaneously deal with these tasks is unaffordable. To this end, we propose an end-to-end unified multi-task learning framework to simultaneously complete the three tasks, where models are trained in an unsupervised manner without using any labels. In particular, we have three parallel branches. In the alignment branch, we adopt a two-stage training scheme, i.e., self-supervised pretraining and constrained unsupervised training using our proposed skip correlation attention layer and constrained loss. Synchronously, in the clustering branch, the learned deep cluster features are utilized to iteratively cluster subtomograms into groups using pseudo-labels from an image-wise Gaussian Mixture Model (GMM). Meanwhile, in the segmentation branch, we use rough pseudo-labels generated from a voxel-wise GMM as supervision signals, and prior knowledge from humans is utilized to jointly learn how to correct these labels as well as predict reliable segmentation results. Benefiting from the end-to-end unified network architecture, our method achieves overall state-of-the-art performance on both simulated and real subtomogram processing benchmarks.

MOTIVATION: Cryo-Electron Tomography (cryo-ET) is a 3D imaging technology that enables the visualization of subcellular structures in situ at near-atomic resolution. Cellular cryo-ET images help in resolving the structures of macromolecules and determining their spatial relationship in a single cell, which has broad significance in cell and structural biology. Subtomogram classification and recognition constitute a primary step in the systematic recovery of these macromolecular structures. Supervised deep learning methods have been proven to be highly accurate and efficient for subtomogram classification, but suffer from limited applicability due to scarcity of annotated data. While generating simulated data for training supervised models is a potential solution, a sizeable difference in the image intensity distribution in generated data as compared with real experimental data will cause the trained models to perform poorly in predicting classes on real subtomograms. RESULTS: In this work, we present Cryo-Shift, a fully unsupervised domain adaptation and randomization framework for deep learning-based cross-domain subtomogram classification. We use unsupervised multi-adversarial domain adaption to reduce the domain shift between features of simulated and experimental data. We develop a network-driven domain randomization procedure with 'warp' modules to alter the simulated data and help the classifier generalize better on experimental data. We do not use any labeled experimental data to train our model, whereas some of the existing alternative approaches require labeled experimental samples for cross-domain classification. Nevertheless, Cryo-Shift outperforms the existing alternative approaches in cross-domain subtomogram classification in extensive evaluation studies demonstrated herein using both simulated and experimental data. AVAILABILITYAND IMPLEMENTATION: https://github.com/xulabs/aitom. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

MOTIVATION: Protein structures provide basic insight into how they can interact with other proteins, their functions and biological roles in an organism. Experimental methods (e.g. X-ray crystallography and nuclear magnetic resonance spectroscopy) for predicting the secondary structure (SS) of proteins are very expensive and time consuming. Therefore, developing efficient computational approaches for predicting the SS of protein is of utmost importance. Advances in developing highly accurate SS prediction methods have mostly been focused on 3-class (Q3) structure prediction. However, 8-class (Q8) resolution of SS contains more useful information and is much more challenging than the Q3 prediction. RESULTS: We present SAINT, a highly accurate method for Q8 structure prediction, which incorporates self-attention mechanism (a concept from natural language processing) with the Deep Inception-Inside-Inception network in order to effectively capture both the short- and long-range interactions among the amino acid residues. SAINT offers a more interpretable framework than the typical black-box deep neural network methods. Through an extensive evaluation study, we report the performance of SAINT in comparison with the existing best methods on a collection of benchmark datasets, namely, TEST2016, TEST2018, CASP12 and CASP13. Our results suggest that self-attention mechanism improves the prediction accuracy and outperforms the existing best alternate methods. SAINT is the first of its kind and offers the best known Q8 accuracy. Thus, we believe SAINT represents a major step toward the accurate and reliable prediction of SSs of proteins. AVAILABILITY AND IMPLEMENTATION: SAINT is freely available as an open-source project at https://github.com/SAINTProtein/SAINT.