Laser microsurgery allows the user to ablate cell bodies or disconnect nerve fibers by using a laser microbeam focused through a microscope. This technique was pioneered in C. elegans where it led to exciting discoveries in the fields of development and neurobiology. All neurons studied so far in C. elegans can regenerate and regrow axons and dendrites after injury, allowing studies of the molecular and cellular basis of neuroregeneration. In this chapter, we describe how to assemble and operate a platform for Yb-doped fiber laser microsurgery. The novel laser setup described here is a more robust, lower cost, and user-friendly alternative to other femtosecond-pulsed laser systems.