High-throughput imaging of neuronal activity in Caenorhabditis elegans

Significance Most behaviors and neuronal responses are variable across individual animals and repeated presentation of the same stimulus. Current neuronal recording techniques examine one animal at a time, whereas hundreds to thousands of trials may be necessary to understand the probability and range of responses. We developed an imaging system to record neuronal activity, detected by genetically encoded calcium indicators, simultaneously from 20 Caenorhabditis elegans animals in microfluidic arenas. We used this system to characterize chemosensory neuron responses to odors and pharmacological manipulation. The system allowed recordings in freely moving animals, whose neuronal responses could be correlated with behavior. We found that behavioral variability is observed even when sensory responses are reproducible, and that sensitivity to specific odors varies among individual animals. Neuronal responses to sensory inputs can vary based on genotype, development, experience, or stochastic factors. Existing neuronal recording techniques examine a single animal at a time, limiting understanding of the variability and range of potential responses. To scale up neuronal recordings, we here describe a system for simultaneous wide-field imaging of neuronal calcium activity from at least 20 Caenorhabditis elegans animals under precise microfluidic chemical stimulation. This increased experimental throughput was used to perform a systematic characterization of chemosensory neuron responses to multiple odors, odor concentrations, and temporal patterns, as well as responses to pharmacological manipulation. The system allowed recordings from sensory neurons and interneurons in freely moving animals, whose neuronal responses could be correlated with behavior. Wide-field imaging provides a tool for comprehensive circuit analysis with elevated throughput in C. elegans.

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