Home › Cell Biology › An In Vivo Duo-color Method for Imaging Vascular Dynamics Following Contusive Spinal Cord Injury
Cell Biology JoVE (Open Access) Citable · DOI

An In Vivo Duo-color Method for Imaging Vascular Dynamics Following Contusive Spinal Cord Injury

DOI: 10.3791/56565-v
What you'll learn
  • ✓ Perform external jugular vein catheterization for fluorescent dye delivery in rats
  • ✓ Install a two-photon imaging window via laminectomy for spinal cord visualization
  • ✓ Acquire baseline and post-injury vascular imaging using dual-color fluorescent dyes
  • ✓ Analyze dynamic vascular changes following contusive spinal cord injury in vivo
Protocol

We introduce an in vivo imaging method using two different fluorescent dyes to track dynamic spinal vascular changes following a contusive spinal cord injury in adult Sprague-Dawley rats.

Difficulty
advanced
Total time
~4–5 hours per animal (surgery, imaging, and recovery)
Model organism
Rat Sprague-Dawley
Biosafety
BSL-1

Steps

1
Catheterize external jugular vein for dye injection

Insert a catheter into the external jugular vein to enable intravenous delivery of fluorescent dyes during imaging. This provides reliable vascular access for both baseline and post-injury injections.

▶ 01:22
2
Stabilize spine and install two-photon imaging window

Perform C5–C7 laminectomy to expose the spinal cord, stabilize the vertebral column, and mount a two-photon imaging window. This enables high-resolution in vivo visualization of spinal vasculature.

▶ 03:09
3
Inject baseline fluorescent dye and acquire images

Administer the first fluorescent dye intravenously and perform baseline two-photon imaging to establish vascular morphology and perfusion patterns before injury.

▶ 05:02
4
Induce contusive injury and image post-injury dynamics

Use the LISA device to deliver a standardized contusive spinal cord injury at C7, inject a second fluorescent dye, and acquire post-injury imaging to visualize acute vascular changes.

▶ 06:25
5
Analyze representative vascular dynamics post-injury

Review and interpret dual-color in vivo imaging data to characterize vascular responses, perfusion alterations, and hemorrhage patterns following contusive spinal cord injury.

▶ 07:26
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