Femtosecond laser nanoaxotomy lab-on-a-chip for in vivo nerve regeneration studies

Abstract

Caenorhabditis elegans is an ideal model organism for studying nerve regrowth and functional recovery after in vivo axotomy, but its high mobility makes such experiments challenging. A microfluidic device capable of transient immobilization of individual worms for high-resolution imaging and laser-based nanoaxotomy is described. A thorough understanding of nerve regeneration in Caenorhabditis elegans requires performing femtosecond laser nanoaxotomy while minimally affecting the worm. We present a microfluidic device that fulfills such criteria and can easily be automated to enable high-throughput genetic and pharmacological screenings. Using the 'nanoaxotomy' chip, we discovered that axonal regeneration occurs much faster than previously described, and notably, the distal fragment of the severed axon regrows in the absence of anesthetics.