Rapid detection and recognition of whole brain activity in a freely behaving Caenorhabditis elegans

Advanced volumetric imaging methods and genetically encoded activity indicators have permitted a comprehensive characterization of whole brain activity at single neuron resolution in Caenorhabditis elegans. The constant motion and deformation of the nematode nervous system, however, impose a great challenge for consistent identification of densely packed neurons in a behaving animal. Here, we propose a cascade solution for long-term and rapid recognition of head ganglion neurons in a freely moving C. elegans. First, potential neuronal regions from a stack of fluorescence images are detected by a deep learning algorithm. Second, 2-dimensional neuronal regions are fused into 3-dimensional neuron entities. Third, by exploiting the neuronal density distribution surrounding a neuron and relative positional information between neurons, a multi-class artificial neural network transforms engineered neuronal feature vectors into digital neuronal identities. With a small number of training samples, our bottom-up approach is able to process each volume—1024 × 1024 × 18 in voxels—in less than 1 second and achieves an accuracy of 91% in neuronal detection and above 80% in neuronal tracking over a long video recording. Our work represents a step towards rapid and fully automated algorithms for decoding whole brain activity underlying naturalistic behaviors.Author summary: An important question in neuroscience is to understand the relationship between brain dynamics and naturalistic behaviors when an animal is freely exploring its environment. In the last decade, it has become possible to genetically engineer animals whose neurons produce fluorescence reporters that change their brightness in response to brain activity. In small animals such as the nematode C. elegans, we can now record the fluorescence changes in and thereby infer neural activity from most neurons in the head of a worm, when the animal is freely moving. These neurons are densely packed in a small volume. Since the brain and body are moving and its shape undergoes significant deformation, a human expert, even after long hours of inspection, may still have difficulty to tell where the neurons are and who they are.