Murine Tail Lymphedema Model: An In Vivo Technique to Generate Sustained Lymphedema in Murine Tail
Transcription
To begin, position the 8-week-old sedated mouse sternally, and prep the tail with 70% isopropyl alcohol. Use a caliper to measure the tail diameter at 5-millimeter increments starting 20-millimeters from the base of the tail. Mark a 3-millimeter circumferential incision on the tail 20-millimeters from the base.
Under surgical microscopic magnification, perform a meticulous 3-millimeter full-thickness skin excision, leaving all the underlying vasculature intact. Incise a superior circumferential mark, first through the dermis, followed by a circumferential full-thickness incision 3-millimeters distal to the first incision. Make a perpendicular full thickness vertical incision to connect the two incisions, then use a toothed fine pickup to grasp a leading edge and dissect deep within the avascular plane to the dermis and superficial to the vein adventitia, with microscissors.
Inject 0.1 milliliter of 1% isosulfan blue subcutaneously proximal to the tip of the tail. Identify the two lymphatic channels appearing blue due to the injection, adjacent to the lateral tail veins. Transect the lymphatics carefully, dissecting a plane between the lateral vein and the lymphatic with straight microsurgical scissors. Pass the tip of one scissor blade between the lymphatic vessel and the lateral vein, and close the blades to transect the lymphatic vessel. When finished, dress the tail wound with a sterile adherent clear dressing.
