Protocol for quantifying Drosophila feeding behavior using flyPAD and optoPAD

Quantifying feeding behavior with precision is essential for understanding how internal state, sensory cues, and neural activity influence food intake and dietary choice. Here, we present a protocol for performing consumption and dietary choice assays in Drosophila using the flyPAD/optoPAD system. We provide instructions for food preparation, arena setup, data acquisition, and analysis for studying feeding behavior, nutrient preference, and learning. This approach enables the measurement of feeding events across multiple arenas while allowing precise control of food substrates and optogenetic stimulation. For complete details on the use and execution of this protocol, please refer to Christie et al.¹.