Home › Neuroscience › Intravital Imaging of Axonal Interactions with Microglia and Macrophages in a Mouse Dorsal Column Crush Injury
Neuroscience JoVE (Open Access) Citable · DOI

Intravital Imaging of Axonal Interactions with Microglia and Macrophages in a Mouse Dorsal Column Crush Injury

DOI: 10.3791/52228-v
What you'll learn
  • ✓ Perform dorsal column crush injury surgery in mice using bone marrow chimera models
  • ✓ Set up and execute two-photon intravital imaging of spinal cord tissue
  • ✓ Visualize and interpret axonal interactions with microglia and macrophages in vivo
Protocol

Biopharma Insights Two-photon intravital imaging can be used to investigate interactions among different cell types in the spinal cord in their native tissue environment in a bone marrow chimeric animal with a dorsal column traumatic spinal cord crush injury.

Difficulty
advanced
Total time
~4–6 weeks (including bone marrow transplantation recovery, injury surgery, and imaging sessions)
Model organism
Mouse (bone marrow chimeric)
Biosafety
BSL-1

Steps

1
Establish dorsal column crush injury model

Perform controlled traumatic crush injury to the dorsal column of the spinal cord in bone marrow chimeric mice. This establishes the injury condition necessary for subsequent imaging of immune cell interactions.

▶ 01:10
2
Prepare tissue and perform intravital imaging

Set up two-photon microscopy imaging system and position the injured spinal cord for live visualization. Acquire high-resolution optical sections to visualize axonal interactions with immune cells in native tissue.

▶ 04:00
3
Capture post-injury cellular interactions

Document and analyze two-photon image sequences showing microglia and macrophage behavior around damaged axons following spinal cord injury.

▶ 06:51
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