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For neuronal tracing, adult brain nerves were dissected in physiological saline based on onychophoran blood composition [ 61].
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Brains, spinal cords and optic nerves were dissected in PBS, fixed with 4% PFA overnight at 4°C, cryoprotected in 30% sucrose overnight at 4°C, then frozen in a 1∶1 mixture of 30% sucrose:OCT (Sakura, CA).
Eyes with optic nerves were isolated from the brain and post-fixed for 2 h in PFA at 4°C, and nerves were dissected and further fixed overnight in 1.2% PFA and 0.8% glutaraldehyde in PB (Electron Microscopy Sciences).
Sciatic nerves were dissected at the level where the nerve exits the greater sciatic foramen.
Mice were perfused with 4 % PFA and sciatic nerves were dissected.
After measuring NCV, the animals were killed, and the sciatic nerves were dissected and flash frozen.
The optic nerves were dissected and immediately fixed in 4% PFA in paraformaldehyde for 1 hour.
Optic nerves were dissected, fixed in 2% osmium tetroxide, and embedded in epon resin.
Nerves were dissected at the indicated days for analysis by immunostaining or EM.
After 30 days, optic nerves were dissected and immediately frozen in liquid nitrogen.
Briefly, sciatic nerves were dissected out and stripped free of epineurium.
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