Robust tracking and quantification of <i>C. elegans</i> body shape and locomotion through coiling, entanglement, and omega bends

The behavior of the well-characterized nematode, Caenorhabditis elegans (C. elegans), is often used to study the neurologic control of sensory and motor systems in models of health and neurodegenerative disease. To advance the quantification of behaviors to match the progress made in the breakthroughs of genetics, RNA, proteins, and neuronal circuitry, analysis must be able to extract subtle changes in worm locomotion across a population. The analysis of worm crawling motion is complex due to self-overlap, coiling, and entanglement. Using current techniques, the scope of the analysis is typically restricted to worms to their non-occluded, uncoiled state which is incomplete and fundamentally biased. Using a model describing the worm shape and crawling motion, we designed a deformable shape estimation algorithm that is robust to coiling and entanglement. This model-based shape estimation algorithm has been incorporated into a framework where multiple worms can be automatically detected and tracked simultaneously throughout the entire video sequence, thereby increasing throughput as well as data validity. The newly developed algorithms were validated against 10 manually labeled datasets obtained from video sequences comprised of various image resolutions and video frame rates. The data presented demonstrate that tracking methods incorporated in WormLab enable stable and accurate detection of these worms through coiling and entanglement. Such challenging tracking scenarios are common occurrences during normal worm locomotion. The ability for the described approach to provide stable and accurate detection of C. elegans is critical to achieve unbiased locomotory analysis of worm motion.