To begin, take the harvested dorsal root ganglia. On a clean bench, fill a 60-millimeter Petri dish with trimming media. Arrange the dissecting scope and glass bead sterilizer, and place an autoclave toolbox with trimming tools next to the scope.
Using the dissecting scope, remove any excess tissue around the dorsal root ganglion, and trim the nerve roots close to the dorsal root ganglion body. Now, fill a second 60-millimeter Petri dish with fresh media. Using the dissecting scope, cut the trimmed dorsal root ganglia to approximately 0.5 millimeters.
Transfer the cut dorsal root ganglia into a fresh 24-well plate containing media on ice. Then, pipette 65.1 and 52.8 microliters of the premixed hydrogel into the soma and neurite compartments, respectively. Carefully embed the cut dorsal root ganglia into the soma compartment.
Thermally cross-link the hydrogel in the incubator at 37 degrees Celsius for 30 minutes. Then, UV cross-link the hydrogel for 90 seconds. Afterward, add dorsal root ganglia growth media to the hydrogel and dorsal root ganglia.
Perform a complete media change after one hour to remove any unreactive or residual photo-initiator in the media.
