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Immunology JoVE (Open Access) Citable · DOI

Laparoscopic Technique for Serial Collection of Liver and Mesenteric Lymph Nodes in Macaques

DOI: 10.3791/55617-v
What you'll learn
  • ✓ Perform laparoscopic entry and cannula placement in macaques
  • ✓ Execute serial mesenteric lymph node biopsies via minimally invasive technique
  • ✓ Collect liver biopsies and identify leukocyte populations from samples
Protocol

Biopharma Insights Here, we describe a minimally invasive laparoscopic technique for serial sampling of liver and mesenteric lymph nodes (MLN) in macaques that allows for increased sampling frequency, and reduces the potential for surgical complications when compared to performing a laparotomy.

Difficulty
advanced
Total time
~2–3 hours per animal (including anesthesia prep and recovery)
Model organism
Macaque (Macaca mulatta or related species)
Biosafety
BSL-2

Steps

1
Prepare animal and surgical field

Anesthetize macaque, position for laparoscopy, and prepare sterile surgical field. Establish vascular access and monitoring as needed for the procedure.

▶ 01:12
2
Establish laparoscopic access and place cannula

Create pneumoperitoneum, insert laparoscope, visualize abdominal cavity, and position working cannulae for lymph node and liver access.

▶ 01:49
3
Biopsy mesenteric lymph nodes

Identify mesenteric lymph nodes under laparoscopic visualization and collect tissue samples using biopsy forceps through working cannula.

▶ 04:17
4
Collect liver tissue biopsy

Locate liver under direct laparoscopic view and obtain biopsy sample from accessible liver margin using biopsy instrument.

▶ 06:38
5
Analyze leukocyte and lymphocyte populations

Process mesenteric lymph node and liver biopsies to identify and characterize leukocyte subsets via flow cytometry or immunohistochemistry.

▶ 07:53
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