To begin, place the 30 gauge one milliliter insulin syringes in a four degrees Celsius refrigerator for cooling. Keep the freshly prepared matrix gel mix containing vascular cells on ice until the mouse is ready for the injection. Next, place the properly anesthetized mouse on the operating board in a prone position and secure its limbs onto the board using tape.
Load a pre cooled insulin syringe with 300 microliters of the matrix gel mixture, avoiding the formation of bubbles. Disinfect the skin on the back of the mouse using 75%alcohol pads, then subcutaneously inject 300 microliters of the matrix gel mixture in the syringe into one side of the animal's back. Gently remove the needle from the injection site to prevent any leakage of the gel mixture.
Ensure the formation of a small hump at the injection site. Position the mouse on a heating pad for two minutes to allow the matrix gel mixture to coagulate and form a plug. Repeat the demonstrated steps to create a plug on the other side of the mouse's back.
Mark the edges of the hump using a marker pen. House the animal in a specific pathogen free experimental animal laboratory at 20 to 24 degrees with 12 hour light and 12 hour dark cycles for seven days.
