Paired-surface spatial mechanomics links tissue stiffness maps to spatial transcriptomics

Tissue mechanics influence diverse biological processes, yet directly linking stiffness measurements to spatially resolved molecular states in intact tissues remains challenging. Here we developed a paired-surface spatial mechanomics approach to map Young's modulus by nanoindentation on a fresh tissue surface and co-register the stiffness grid with Visium HD spatial transcriptome data from the immediately adjacent, parallel surface. This was applied to the mouse ovary, which has spatially distinct compartments and undergoes extracellular matrix remodeling with age. Nanoindentation at 50 m grid spacing enabled millimeter-scale stiffness maps while balancing acquisition time in fresh tissues, generating 2,771 matched measurements across 21 regions of interest. Global and compartment-specific analyses associated stiffer regions with lower matrix-related programs and higher cell-cycle programs, alongside age-dependent inflammatory and metabolic associations. This correlative strategy integrates experimentally measured mechanics with spatial omics in fresh tissues.