SparseSeg: Target-Conditioned Discovery Segmentation of Cryo-Volume Electron Microscopy Under Sparse Annotation

Cryo-volume electron microscopy (cryo-vEM) enables near-native visualization of cellular ultrastructure, but its broad use is limited by low image contrast and the high cost of dense voxel-level annotation. Existing automated segmentation methods often generalize poorly across cell types, organelles, and imaging conditions. Here, we introduce SparseSeg, a target-conditioned, sparsity-driven segmentation framework that treats organelle segmentation as a discovery process rather than a closed-set classification task. SparseSeg uses a small number of context-specific exemplars to iteratively propagate reliable supervision through the volume. It combines sparse patch-based sampling, a multi-kernel U-Net, and geometry-consistent refinement to expand accurate segmentation while suppressing context-dependent false positives. Across serial cryo-FIB–SEM and conventional vEM datasets, SparseSeg achieves robust segmentation under extreme sparse annotation, including settings with less than 1% labeled slices. This framework reduces annotation burden while preserving morphological fidelity for quantitative cryo-vEM analysis.