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.
Chakraborty, S., van Veghel, M., Tzanou, A., Li, Z., Torbin, D., Lowet, E.
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