In situ graphene-seq: spatial transcriptomics and chronic electrophysiological characterization of tissue...

Biological systems comprise diverse, interconnected cell types whose functional dynamics and molecular identities are tightly coupled, yet difficult to capture simultaneously at high spatiotemporal resolution. Electrophysiology provides real-time measurements of cellular activity but with limited molecular context, whereas transcriptomics profiles gene expression without dynamic physiological readouts. Here, we introduce in situ graphene-sequencing, a platform that integrates chronic electrophysiology with imaging-based, spatially resolved transcriptomics (STARmap). The system combines stretchable mesh nanoelectronics for long-term, single-cell-level interfacing with transparent graphene/poly(3,4-ethylenedioxythiophene) polystyrene sulfonate electrodes, enabling seamless integration of electrical recording and optical imaging. By coupling electrophysiology with STARmap, the platform enables multimodal analysis of heterogeneous tissue microenvironments. We demonstrate in situ graphene-sequencing by charting multimodal profiles of human-induced pluripotent stem cell-derived cardiomyocyte and endothelial cell co-cultures, examining how spatial heterogeneity is associated with electrophysiological activity and gene expression. This approach provides an integrative framework for studying how tissue microenvironments shape cell behavior and molecular states.