Home › Microscopy & Imaging › Visualizing Leukocyte Rolling and Adhesion in Angiotensin II-Infused Mice: Techniques and Pitfalls
Microscopy & Imaging JoVE (Open Access) Citable · DOI

Visualizing Leukocyte Rolling and Adhesion in Angiotensin II-Infused Mice: Techniques and Pitfalls

DOI: 10.3791/56948-v
What you'll learn
  • ✓ Implant jugular catheters and prepare carotid arteries for intravital imaging
  • ✓ Perform intravital video microscopy to visualize leukocyte rolling and adhesion
  • ✓ Quantify immune cell activation in angiotensin II-induced hypertension models
  • ✓ Troubleshoot common technical pitfalls in leukocyte imaging protocols
Protocol

This manuscript describes the use of transgenic reporter mice and different administration routes of fluorescent dyes in angiotensin II-induced hypertension using intravital video microscopy of blood vessels to evaluate the activation of immune cells and their ability to roll and adhere to the endothelium.

Difficulty
advanced
Total time
~4–6 hours per mouse (surgery + imaging + analysis)
Model organism
Mouse (transgenic LysM-GFP or similar reporter strain)
Biosafety
BSL-1

Steps

1
Implant jugular catheter and prepare carotid vessels

Surgically implant a jugular catheter for angiotensin II infusion and expose carotid arteries for intravital microscopy imaging. This establishes vascular access and visualization field for downstream leukocyte tracking.

▶ 00:51
2
Acquire intravital video microscopy of rolling and adherent leukocytes

Use fluorescent dye administration and intravital microscopy to visualize and record leukocyte rolling and adhesion dynamics on the endothelium in real time. Document cell behavior at multiple vessel sites for quantitative analysis.

▶ 05:11
3
Analyze leukocyte rolling and adhesion in angiotensin II conditions

Quantify rolling velocity, adhesion frequency, and LysM+ cell activation in angiotensin II-infused versus control mice. Compare results across experimental groups to assess immune cell engagement with inflamed endothelium.

▶ 07:29
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