Figure 12.

Surgeries for implanting terminal windows in the mouse popliteal lymph node

Left panel: Preparation of surgical site. Begin by injecting anesthetic intraperitoneally. Diagram and picture show site of incision. Make a small incision on one of the lower limbs (up to 2 mm) and carefully move the subcutaneous adipose tissues and muscles to expose the popliteal lymph node. Middle panel: Place window for imaging. Two options exist for imaging the popliteal lymph node either with or without a coverslip. Option 1: Hydrate the site of incision and image directly. Option 2: Use a round coverslip attached to a vacuum ring and place on top of the incision. Right panel: Place mouse on stage and image. A custom-built stage provides full body support to the mouse and allows space for tubing (used for vacuum, and intravenous cannulation). Stretch the leg fully and immobilize all limbs and tail. For stabilization of the leg, a lumbar and hip clamp must be used. Depending on the microscope, an additional custom-built support for the leg might be required. Depending on the experimental conditions, dyes can be added intravenously either before or after securing the mouse on the stage. The objective reaches the lymph node from the top. Scale bar: 20 μm. Created with BioRender.com.