Epidermal Laser Wounding: Inducing Localized Damage to Examine Repair Processes in C. elegans

- To begin, transfer young adult worms onto a fresh agarose pad on a slide. Then add a few drops of 12 millimolar levamisole solution to paralyze the worms. Cover the worms with a coverslip and wait for a few minutes. Place the slide under a spinning disk confocal microscope and focus on the lateral outermost surface of the worm.

The epidermis of an adult worm is a thin single-layered structure consisting of multi-nucleated syncytia that result from cell fusion events. The apical surface of epidermis secretes a flexible collagenous cuticle, while the basal surface is covered by a thin layer of extracellular matrix called the basal lamina.

Next, wound the apical surface of the epidermis with a laser. Cytoplasm from the damaged cells may leak out, appearing as a bubble at the wound site.

Laser irradiation has an advantage over other techniques, such as needle wounding, as it causes localized disruption of the cuticle and epidermal cells, without damaging internal tissues. In the example protocol, we will see the process of laser wounding in C. elegans.

- To precisely wound the nematodes, this protocol uses a femtosecond laser attached to spinning disk confocal microscope. Line or poi