AI Article Synopsis
- A protocol for spinal cord laminectomy and glass window implantation is outlined, enabling in vivo imaging of the mouse spinal cord using multi-photon fluorescence microscopy.
- An integrated digital vaporizer ensures stable anesthesia with low-flow isoflurane, while the procedure involves removing a vertebra, placing a cover-glass, and securing a backplate to minimize motion artifacts.
- Applications of the technique are demonstrated with two-photon microscopy showcasing spinal cord vasculature in transgenic mice expressing eGFP:Claudin-5.
Article Abstract
This protocol describes a method for spinal cord laminectomy and glass window implantation for in vivo imaging of the mouse spinal cord. An integrated digital vaporizer is utilized to achieve a stable plane of anesthesia at a low-flow rate of isoflurane. A single vertebral spine is removed, and a commercially available cover-glass is overlaid on a thin agarose bed. A 3D-printed plastic backplate is then affixed to the adjacent vertebral spines using tissue adhesive and dental cement. A stabilization platform is used to reduce motion artifact from respiration and heartbeat. This rapid and clamp-free method is well-suited for acute multi-photon fluorescence microscopy. Representative data are included for an application of this technique to two-photon microscopy of the spinal cord vasculature in transgenic mice expressing eGFP:Claudin-5 - a tight junction protein.
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| http://dx.doi.org/10.3791/58330 | DOI Listing |
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