Bridging the Dimensional Gap from Planar Spatial Transcriptomics to 3D Cell Atlases

Abstract Spatial transcriptomics (ST) has revolutionized our understanding of tissue architecture, yet constructing comprehensive three-dimensional (3D) cell atlases remains challenging due to technical limitations and high cost. Current approaches typically capture only sparsely sampled two-dimensional sections, leaving substantial gaps that limit our understanding of continuous organ organization. Here, we present SpatialZ, a computational framework that bridges these gaps by generating virtual slices between experimentally measured sections, enabling the construction of dense 3D cell atlases from planar ST data. SpatialZ is designed to operate at single-cell resolution and function independently of gene coverage limitations inherent to specific spatial technologies. Comprehensive validation using real 3D ST and independent serial sectioning datasets demonstrates that SpatialZ accurately reconstructs virtual slices while preserving cell identities, gene expression patterns, and spatial relationships. Leveraging the BRAIN Initiative Cell Census Network data, we constructed a 3D hemisphere atlas comprising over 38 million cells, a scale not feasible experimentally. This dense atlas enables unprecedented capabilities, including in silico sectioning at arbitrary angles, explorations of gene expression across both 3D volumes and surfaces, and 3D mapping of query tissue sections. While currently validated for spatial transcriptomics, the underlying principles of SpatialZ could potentially be adapted for spatial proteomics, spatial metabolomics, and even spatial multi-omics. Validated through internal and external testing, our computationally generated atlas maintains biological accuracy, providing unprecedented resolution of spatial molecular landscapes and demonstrating the potential of computational approaches in advancing 3D ST.