Two-Photon Laser Axotomy: A Method to Injure Axons in Zebrafish Embryos and Observe Axonal Recovery
Published: April 30, 2023
Abstract
Source: O'Brien G. S., et al. Two-photon axotomy and time-lapse confocal imaging in live zebrafish embryos. J. Vis. Exp. (2009).
This video describes the method to injure axons in zebrafish embryos using two photon laser axotomy and observing axonal recovery from injury.
Protocol
1: Two-photon axotomy using a custom built two-photon microscope with a Chameleon Ti-Sapphire laser Prepare for imaging. Place the mounted embryo(s) on a slide holder under the microscope. Focus on one embryo with a 40X (0.8 NA) water objective. Turn on the laser. We have been able to reproducibly visualize and damage GFP expressing neurons using the following parameters. To visualize axons at a non-damaging power, set laser to a wavelength of 910 nanometers (nm) and a power of 30 milliwatts (mW listed are th…
Cite This Article
Two-Photon Laser Axotomy: A Method to Injure Axons in Zebrafish Embryos and Observe Axonal Recovery. J. Vis. Exp. (Pending Publication), e20188, doi: (2023).