Surface Acoustic Wave Integrated Microfluidics for Repetitive and Reversible Temporary Immobilization of C. elegans

Caenorhabditis elegans is an important genetic model for neuroscience studies due to its unique combination of genetics, transparency, complete synaptic connectome, and well-characterized behaviors. These factors, in turn, enable analyses of how genes control connectivity, neuronal function, and behavior. To date, however, most studies of neuronal function in C. elegans are incapable of performing microscopy imaging with subcellular resolution and behavior analysis in the same set of animals. This constraint stems from the immobilization requirement for high-resolution imaging that is incompatible with behavioral analysis. In particular, conventional immobilization methods often lead to either irreversible, partial, or slowly reversible immobilization of animals preventing a multiplexed approach. Here, we present a novel microfluidic device that uses surface acoustic waves (SAW) as a non-contact method to temporarily immobilize worms for a short period (40 seconds). This device allows non-invasive analysis of swimming behavior and high-resolution synaptic imaging in the same animal. In addition, because of the low impact of this SAW approach, the device enables fast, repeated imaging of single neurons and behavior in the same animals for three to four days. We anticipate that this device will enable longitudinal analysis of animal motility and subcellular morphological changes during development and ageing in C. elegans .