Semi-supervised clustering with knowledge-guided representation learning in cryo-electron tomography.

The automated discovery of structural patterns in macromolecular complexes remains a central challenge in cryo-electron tomography, particularly in highly heterogeneous datasets. Although fully unsupervised clustering methods have shown promise in grouping subtomograms by structural similarity, they often ignore a crucial source of information: the partial ground truth routinely available to structural biologists from prior studies or manual annotations. In this work, we propose a semi-supervised structural discovery framework that utilizes partial supervision to guide clustering without compromising the ability to uncover previously unknown structures. At the core of our method is a label-anchored probabilistic clustering mechanism that seeds the latent space using a small subset of labeled examples and refines it through a multi-resolution consensus strategy based on PCA-space voting. This is complemented by an entropy-based confidence scoring scheme that attenuates the influence of ambiguous samples, as well as a feature propagation procedure that extends structural labels to low-confidence regions using local similarity in feature space. Together, these components create a stable and adaptive pipeline capable of discovering both known and novel structures. Our approach is efficient, requires as little as 1% of labeled data per class, and consistently produces clearer, more interpretable feature embeddings compared to fully unsupervised methods, with well-separated clusters from the very first iterations. Extensive experiments on simulated and realistic tomographic datasets demonstrate that this semi-supervised strategy significantly improves clustering performance, robustness, and biological relevance in cryo-electron tomography analysis. These methods are integrated as extensions to the existing Deep Iterative Subtomogram Clustering Approach pipeline, enhancing its capability for guided structural discovery.