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

Area of Science:

• Biotechnology
• Bioelectronics
• Systems Biology

Background:

• Biological systems feature complex, interconnected cell types with coupled dynamics and molecular identities, challenging to study at high spatiotemporal resolution.
• Current methods like electrophysiology lack molecular context, while transcriptomics miss dynamic physiological data.
• Simultaneously capturing cellular activity and molecular profiles is crucial for understanding tissue microenvironments.

Purpose of the Study:

• To introduce a novel platform, in situ graphene-sequencing, that integrates chronic electrophysiology with imaging-based, spatially resolved transcriptomics.
• To enable multimodal analysis of heterogeneous tissue microenvironments by combining electrical recording and optical imaging.
• To investigate the relationship between spatial heterogeneity, electrophysiological activity, and gene expression in co-cultured cells.

Main Methods:

• Development of a platform combining stretchable mesh nanoelectronics with transparent graphene/poly(3,4-ethylenedioxythiophene) polystyrene sulfonate electrodes.
• Integration of long-term, single-cell-level electrophysiological recording with high-throughput, imaging-based in situ sequencing.
• Demonstration using human-induced pluripotent stem cell-derived cardiomyocyte and endothelial cell co-cultures.

Main Results:

• Successful integration of chronic electrophysiology with spatially resolved transcriptomics.
• Charting of multimodal profiles in cell co-cultures, revealing associations between spatial heterogeneity, electrical activity, and gene expression.
• Demonstration of the platform's capability to analyze heterogeneous tissue microenvironments at single-cell resolution.

Conclusions:

• In situ graphene-sequencing provides an integrative framework for studying complex biological systems.
• The platform enables simultaneous measurement of cellular electrophysiology and gene expression, bridging a critical gap in current technologies.
• This approach facilitates a deeper understanding of how tissue microenvironments influence cellular behavior and molecular states.