Tracking multi-site somatic voltage dynamics via high-speed fiber photometry

Abstract

Investigating neural circuit dynamics across distributed brain regions in awake, behaving animals is crucial for understanding complex behavior. Genetically encoded voltage indicators (GEVIs) offer a powerful approach to tracking transmembrane voltage with high temporal and cellular specificity. However, scaling high-sensitivity GEVI recordings across multiple brain regions and multiple animals simultaneously remains a major technical challenge. Furthermore, it is unclear whether soma-targeted GEVIs - typically used for single-cell resolution imaging - can be effectively adapted for fiber photometry. Here, we show that a sCMOS-based widefield imaging system achieves sensitive dual-color multi-site fiber photometry using soma-targeted GEVI indicators with high temporal resolution. We validated this approach in the mouse hippocampal CA1, capturing theta (3-10Hz) and gamma (30-80Hz) rhythms and theta-gamma cross-frequency coupling. Additionally, we recorded high-frequency neural entrainment (>100 Hz) and somatic depolarization induced by electrical stimulation in CA1. Lastly, we tracked synchronized neural activity between the bilateral CA1s as well as multi-site dual-color imaging across CA1 and cortex simultaneously in three freely running mice. This work provides a scalable, accessible platform for high-speed optical electrophysiology in distributed neural circuits. Key points- Implementation of a sCMOS-based widefield imaging setup for sensitive and scalable fiber photometry and cellular imaging. - Demonstration of population multi-site and inter-animal voltage imaging with soma-targeted genetically encoded voltage indicators. - Tracking of high-frequency gamma and high-gamma (>100Hz) neural entrainment