All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Mice used for the experiment
<ol>
	<li>Use 8-week-old male and female mice on the B6 background.</li>
</ol>
2. Surgery procedure to remove the inguinal draining lymph node
<ol>
	<li>Euthanize mice using CO2 asphyxiation followed by cervical dislocation.</li>
	<li>Immobilize mice on the acrylic stage with adhesive tape and apply mineral oil with a cotton swab to the abdominal skin to prevent fur deposition around the incision (Figure 1B). Fur removal is not necessary.</li>
	<li>Perform a midline incision using microsurgery curved scissors (11.5 cm) and microsurgery curved forceps (12.5 cm) in the abdomen from the pubis to the xiphoid process (Figure 1C).</li>
	<li>Dissociate the abdominal musculature from the skin (Figure 1D).</li>
	<li>Make horizontal skin incisions at the top and bottom of the vertical incision line to create skin flaps on the side of interest (according to the side of the antibody mix injection) and flap the skin to visualize the lymph node (Figure 1E).</li>
	<li>Tape the skin flap on the acrylic plate (Figure 1F).</li>
	<li>Remove the inguinal draining lymph node using microsurgery curved forceps (Figure 1F). Lymph nodes will appear as translucid, usually bilobular, spheres under the skin.</li>
</ol>

<table><tbody><tr><td>Microsurgery curved forceps</td><td>WEP Surgical Instruments</td><td>Custom-made</td><td>12.5 cm</td></tr><tr><td>Microsurgery curved scissors</td><td>WEP Surgical Instruments</td><td>Custom-made</td><td>11.5 cm</td></tr></tbody></table>

harvesting an inguinal draining lymph node from a murine model

Harvesting an Inguinal Draining Lymph Node from a Murine Model

Source: Rezende, R. M. et al., <a href='https://app.jove.com/v/59335'>Visualizing Lymph Node Structure and Cellular Localization using Ex-Vivo Confocal Microscopy.</a> J. Vis. Exp. (2019)
This video demonstrates the surgical isolation procedure of the inguinal draining lymph node from a mouse model. Incision of the abdominal region provides access to the inguinal lymph node, which is surgically harvested. The harvested translucent inguinal lymph node can be used for immunological studies.

<img alt="Animal surgery procedure; tissue isolation, biopsy prep, fluorescent imaging setup; research method." src="/files/ftp_upload/21577/21577_Figure1.jpg" data-altupdatedat="1753371239000" data-alt="Mouse surgery procedure; tissue extraction, handling, and analysis, UV illumination step." /> 
Figure 1: Inguinal lymph node preparation. 
(A) Subcutaneous injection of FACS antibody master mix into the inner thigh. (B) 3 h after the injection, euthanize the ...

Source: Rezende, R. M. et al., Visualizing Lymph Node Structure and Cellular Localization using Ex-Vivo Confocal Microscopy.&#160;J. Vis. Exp.&#160; ...

In mice, the inguinal lymph nodes are small, ovoid-shaped organs of the lymphatic network located in the inguinal region &#8212; the area between the thighs and the lower abdomen.
The inguinal lymph nodes receive lymphatic drainage from the mouse's lower extremities. It filters the lymph to identify pathogens and cancerous cells to mount an immune response.
To harvest a murine inguinal draining lymph node, prepare a euthanized mouse in the supine position and secure it on an acrylic stage. Apply mineral oil on the inguinal region to avoid fur interference during the incision.
Make a longitudinal incision along the abdominal area, from the pubis to the xiphoid process &#8212; the lower end of the sternum, exposing the inguinal region.
Next, dissociate the skin from the abdominal muscles for efficient extraction. Create small horizontal incisions at the top and bottom of the longitudinal incision, creating a skin flap.
Retract the skin flap, and secure it to access the underlying tissues, including the inguinal lymph node. Pierce through connective tissue to access the lymphatic system and extract the inguinal lymph node from the system.
The harvested bi-lobular inguinal lymph node is ready for future experiments.

For inguinal draining lymph node harvest, apply mineral oil to the abdominal skin. Use standard microsurgery scissors and microsurgery curved forceps to make a midline skin incision in the abdomen from the pubis to the xiphoid process, and dissociate the abdominal musculature from the skin.
Make horizontal skin incisions at the top and bottom of the vertical incision line to create skin flaps on the side of injection, and retract the skin to visualize the lymph node. Secure the skin flap to the acrylic plate, and use the forceps to remove the translucid, usually bilobular, spherical inguinal draining lymph node.

harvesting-an-inguinal-draining-lymph-node-from-a-murine-model

Research

JoVE Encyclopedia of Experiments

Immunology

Immune Systems and Components

Cell Isolation and Culture

6.4K Views. In mice, the inguinal lymph nodes are small, ovoid-shaped organs of the lymphatic network located in the inguinal region — the area between the thighs and the lower abdomen. The inguinal lymph nodes receive lymphatic drainage from the mouse's lower extremities. It filters the lymph to identify pathogens and cancerous cells to mount an immune response. To harvest a murine inguinal draining lymph node, prepare a euthanized mouse in the supine position and secure it on an acr...

Abstract

Protocol

<img title="figure-results-1" alt="Animal surgery procedure; tissue isolation, biopsy prep, fluorescent imaging setup; research method." src="/files/ftp_upload/21577/21577_Figure1.jpg" data-altupdatedat="1753371239000" data-alt="Mouse surgery procedure; tissue extraction, handling, and analysis, UV illumination step." /> 
Figure 1: Inguinal lymph node preparation. 
(A) Subcutaneous injection of FACS antibody master mix into the inner thigh. (B) 3 h after the injection, euthanize the ...

Harvesting an Inguinal Draining Lymph Node from a Murine Model

In This Article

Abstract

Protocol

Results

Disclosures

Materials

Tags

Name	Company	Catalog Number	Comments
Microsurgery curved forceps	WEP Surgical Instruments	Custom-made	12.5 cm
Microsurgery curved scissors	WEP Surgical Instruments	Custom-made	11.5 cm