Secure an anesthetized mouse bearing an implanted tumor onto a stereotactic frame.
Remove the staples from the previous surgery, disinfect the skin, and incise along the previous wound to expose the skull.
Locate the burr hole from the previous surgery to access the brain.
Position the minimally invasive resection system, or MIRS, over the burr hole.
The system consists of a cannula with a rotating window that uses suction to draw the targeted tissue and a cylindrical blade at the aperture to excise the tissue.
Insert the cannula into the target region and initiate resection.
The suction pulls small tumor sections through the window, while the blade excises the tissue.
The rotating window allows controlled resection via continuous suction and cutting.
The excised fragments travel through a sterile tubing to the sample collection chamber.
Withdraw the cannula, close the wound, and allow the mouse to recover.
Begin the experiment by assessing the mouse for sedation by pinching the toe. Apply ophthalmic ointment to the eyes to avoid dryness of the cornea. Then, place the mouse on a stereotactic frame. Remove the staple from the previous surgery and disinfect the skin with alternating cycles of a chlorhexidine or betadine-based scrub and alcohol. Then create a 1 centimeter longitudinal midline incision along the previous surgical scar using a sterile scalpel. Attach the MIRS handpiece to the stereotactic arm through the stage adapter.
To set up the MIRS machine, insert the power cord set on the rear panel into the power cord receptacle. Turn the power to the system on or off by toggling between 0 and 1. Insert one end of the nitrogen hose into the male fitting on the rear panel of the console. Rotate the connection nut clockwise to tighten it. Ensure that the supply pressure does not exceed 100 PSIG, and attach the hose to the nitrogen supply.
Seal the lid of the vacuum port to avoid any leakage. Check the aspiration knob on the front of the console is set at 100, but there is no leakage in the aspiration system and that the nitrogen input supply pressure is correct. Insert the gray foot pedal connector into its gray receptacle until it clicks. In the same way, insert the blue handpiece connector into its blue receptacle. Prime each handpiece by aspirating sterile fluid into the aperture through the tubing and handpiece, and then into the canister to ensure that the inside of the tubing and handpiece are lubricated.
Select the mode for aspiration on the console front panel and initiate using the foot pedal. Insert the 23G MIRS cannula into the borehole to a depth of 2.5 millimeters. Initiate the resection process by pressing the foot pedal connected to the cannula. Perform full cycles of resection using the control knob in the handpiece. After the resection process, withdraw the 23 gauge MIRS cannula and add 5 milliliters of 1x PBS to flush the tubing and dislodge any residual debris. Then close the wound with a stapler and remove the mouse from the stereotactic frame. Return the mouse to the heating pad to recover from anesthesia before placing it back into its cage.
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