Single-cell spatial multi-omics molecular pathology enabled by SuperFocus

Histopathology and molecular pathology are currently distinct diagnostic modalities for the most part, one revealing tissue morphology at cellular resolution and the other providing molecular measurements with limited or no spatial context. Projecting genome-scale molecular information onto histopathology images at single-cell resolution across whole tissue sections represents a long-sought goal for next-generation pathology. Here we present SuperFocus, a modality-agnostic computational platform that generates histopathology-integrated single-cell spatial multi-omics from spot-based spatial measurements acquired on the same or an adjacent section without requiring external reference data. SuperFocus combines constrained cascading imputation with feature-level and cell-level quality-control scores to reduce spurious predictions and quantify confidence. On a ground-truth spatial transcriptomics benchmark dataset, SuperFocus improves key accuracy metrics by 28-73% over existing methods. Across Patho-DBiT, spatial ATAC-RNA, spatial CITE-seq and Visium-MALDI-MSI (SMA) datasets, SuperFocus enables cell-resolved analyses of MALT lymphoma microenvironments, gene regulatory programs in human hippocampus, lipotoxic hepatocyte states in human MASH, and transcriptomic-metabolomic states linked to neurotransmission and neuroinflammation in Parkinsonian mouse brain. Overall, SuperFocus enables scalable whole-slide single-cell spatial multi-omics integrated with histopathology, bridging histology and genome-scale molecular profiling for next-generation molecular pathology.