Long-term culture and high-resolution imaging of C. elegans using an automated microfluidic platform

The nematode Caernohabditis elegans is one of the most employed small model organisms in biology. Studies on transgenic animals often require the accurate observation of highly localized fluorescent signals inside the worms at high magnification, hence demanding the full immobilization of the animals. Moreover, in order to observe the dynamics of biological processes over the lifespan of a worm, the same worm has to be immobilized repeatedly, in a reversible manner and under normal physiological conditions. We describe a microfluidic platform for the automated culture, treatment and long-term high-resolution imaging of C. elegans. Our device features: (i) a microfluidic design tailored for the isolation of L4 larvae from a mixed larval population and for their successive culture and treatment; (ii) a worm immobilization method, based on the thermoreversible sol-gel transition of the biocompatible triblock copolymer Pluronic F127 inside the microfluidic chip, thereby enabling high-resolution imaging; (iii) an integrated temperature control system, both to ensure viable environmental conditions for C. elegans culture and to steer the worm immobilization/release process. We apply this device to observe mitochondrial dynamics in muscle cells during aging at single worm resolution. We expect our platform to enable the simultaneous study and repeated observation of multiple phenotypes in single worms or specific worm populations, such as lifespan and motility assays, in addition to high-resolution imaging.