An epifluorescence microscope design for naturalistic behavior and cellular activity in freely moving Caenorhabditis elegans.

Abstract

Understanding the neural basis of behavior requires imaging cellular activity in freely moving animals, which typically demands expensive, restrictive microscopy setups. To overcome these barriers, we developed Wormspy, a cost-effective, open-source epifluorescence microscopy system for high-magnification imaging and tracking of Caenorhabditis elegans. Wormspy enables the simultaneous recording of neuronal activity and behavioral dynamics without needing the animal to be restrained. We demonstrate its utility in imaging body wall muscles, sensory neurons, and subcellular calcium events within interneuron axons. Our platform reproduces known mutant phenotypes and uncovers, to the best of our knowledge, previously inaccessible sensorimotor correlations. We show that Wormspy provides a robust, modular framework that lowers technical barriers to high-resolution neural imaging, enabling flexible experimental designs for dissecting behavior in freely moving organisms.