Injection of cholera toxin subunit B in the interscapular brown adipose fat and Stellate Ganglion dissection

Prepare the surgery area by placing a clean paper towel on top of a heat pad

Anesthetize the animal under isoflurane until there's no toe pinch reflex

Transfer the mouse to the surgery area, flat on its stomach on the heatpad. Place the nose cone to maintain anesthesia

Shave the interscapular area of the mouse and sterilize it with three alternating wipes of betadine and 70% isopropyl alcohol

Apply eye ointment

Administer analgesics. We use buprenorphine 0.1 mg/kg subcutaneously

Load up cholera toxin subunit B (CTB) in a glass pulled needle connected to a mouth pipette (10 uL per side)

Make a 1 cm horizontal incision above the interscapular area

Visually identify the white adipose layer of interscapular pad, and use scissors to cut it and expose the underlying brown adipose tissue

While holding the tissue to expose the brown fat, inject 10 uL of CTb on each side

Gently return the fat pads to their original position, and close the wound with a 4-0 PS-4 suture

Clean the wound with betadine and 70% isopropyl alcohol and monitor the animal for any sign of distress

Euthanize the mouse after 72 hours by intraperitoneal injection of 2-2-2-Tribromoethanol (500 mg/kg)

Open the ribcage and remove the lungs for easier access to the sympathetic chain

Carefully dissect the Stellate Ganglion from each side