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Neuroscience JoVE (Open Access) Citable · DOI

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

DOI: 10.3791/52196-v
What you'll learn
  • ✓ Prepare a mouse for chronic spinal cord imaging surgery
  • ✓ Perform dorsal laminectomy to expose the spinal cord
  • ✓ Implant and seal an optical imaging chamber over dorsal spinal cord
  • ✓ Execute longitudinal multi-photon imaging through the chronic chamber
Protocol

In this video, we describe a procedure for implanting a chronic optical imaging chamber over the dorsal spinal cord of a live mouse. The chamber, surgical procedure, and chronic imaging are reviewed and demonstrated.

Difficulty
advanced
Total time
~2–3 hours per mouse (surgical implantation); ongoing imaging sessions ~30–60 min per timepoint
Model organism
Mouse (strain not specified)
Biosafety
BSL-1

Steps

1
Prepare the mouse for surgery

Anesthetize and position the mouse on the surgical platform. Prepare the surgical field and establish asepsis protocols.

▶ 01:56
2
Expose the dorsal laminae of thoracic vertebrae

Make midline incision and retract skin and musculature to access the dorsal spinal column at the thoracic level.

▶ 02:56
3
Perform a dorsal laminectomy

Remove the laminae overlying the dorsal spinal cord using micro-surgical tools to expose the dorsal surface without damaging underlying tissue.

▶ 05:11
4
Seal the chamber over exposed spinal cord

Position the optical imaging chamber over the exposed dorsal spinal cord and secure it with dental cement to create a chronic sealed imaging window.

▶ 06:54
5
Perform chronic multi-photon imaging sessions

Use the implanted chamber to conduct repeated longitudinal in vivo imaging of the spinal cord using multi-photon microscopy at multiple timepoints.

▶ 09:16
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