Simultaneous interneuron labeling reveals cell type-specific, population-level interactions in cortex

Cortical interneurons shape network activity in cell type-specific ways, and interact with other cell types. These interactions are understudied, as current methods typically restrict in vivo labeling to one neuron type. Although post-hoc identification of many cell types has been accomplished, the method is not available to many labs. We present a method to distinguish two red fluorophores in vivo, allowing imaging of activity in somatostatin (SOM), parvalbumin (PV), and the rest of the neural population in mouse cortex. We compared population events in PV and SOM neurons and observed that local network states reflected the ratio of SOM to PV neuron activity, demonstrating the importance of simultaneous labeling to explain dynamics. Activity became sparser and less correlated when the ratio between SOM and PV activity was high. Our simple method can be flexibly applied to study interactions among any combination of distinct cell type populations across brain areas.