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Microscopy & Imaging JoVE (Open Access) Citable · DOI

Intravital Microscopy of the Inguinal Lymph Node

DOI: 10.3791/2551-v
What you'll learn
  • ✓ Prepare and surgically expose the inguinal lymph node for intravital imaging
  • ✓ Acquire real-time in vivo microscopy images of lymph node microvasculature
  • ✓ Analyze vessel morphology and assess tissue viability during imaging
  • ✓ Adapt protocol for cell trafficking and infection studies
Protocol

A technique for performing intravital microscopy of the inguinal lymph node (LN) is outlined. Such technique allows for real-time, in vivo study of the lymph node microvasculature and structure both during homeostasis and infection. This technique can be adapted to cell trafficking studies and to other lymph node sites.

Difficulty
advanced
Total time
~90–120 min/mouse (anesthesia induction, surgical prep, imaging, recovery)
Model organism
Mouse (unspecified strain, likely C57BL/6 or similar)
Biosafety
BSL-1

Steps

1
Prepare animal for anesthesia and surgery

Anesthetize the mouse and position for inguinal lymph node access. Prepare surgical field and animal positioning to enable stable intravital microscopy imaging.

▶ 01:30
2
Surgically expose the inguinal lymph node

Perform incision and dissection to expose the inguinal lymph node while preserving microvasculature and surrounding tissue integrity for imaging.

▶ 01:54
3
Acquire intravital microscopy images and analyze vessels

Capture real-time fluorescence and brightfield images of lymph node microvasculature, measure vessel diameter and blood flow dynamics.

▶ 03:54
4
Verify lymph node and tissue viability post-imaging

Assess vascular perfusion, cellular morphology, and metabolic markers to confirm the lymph node remained viable throughout intravital microscopic evaluation.

▶ 05:57
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