During C. elegans lethargus, a sleep-like behavior called quiescence occurs in short bouts during which animals spontaneously pause locomotion and feeding (Raizen et al., 2008). Multiple laboratories are interested in studying quiescence, however monitoring this behavior is problematic as highmagnification images must be captured for individual animals over a long period of time. Using a dissection scope with a camera to measure quiescence is generally expensive and monopolizes microscopes that would normally be used for other purposes. We describe here a set-up that can be used to follow individual animals without using a dissection microscope. We load up to 6 late-L4 animals into simple, reusable microfluidic chambers and analyze images captured every 10 seconds throughout lethargus (Singh et al., 2011). MATLAB scripts for acquiring and analyzing the images are available upon request.