Source: Geisler, J. A. et al. Modeling Brain Metastases Through Intracranial Injection and Magnetic Resonance Imaging. J. Vis. Exp. (2020)
In this video, we describe the surgical procedure to create a calvarial window for accessing the brain of a mouse model.
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Prepare Stereotactic Units for Surgery
2. Place Mice on Stereotactic Frames
3. Make a Calvarial Window
Figure 1: Stereotactic and anesthesia systems. (A) Anesthesia induction chamber setup within a biological safety cabinet. (B) Stereotactic anesthetic delivery setup highlighting anesthesia tubing from the anesthetic machine to the nose cone on the stereotactic apparatus (see inset in (C)). Green arrows indicate delivered anesthetic gas tubing, and blue arrows indicate scavenged gas tubing. (C) Stereotactic stand with drill attachment. Inset shows anesthetic tubing (green and blue arrows), mouth bar, and ear bars. (D) Stereotactic stand with automated syringe pump and mouse elevating box. The box is an inverted pipette tip box containing hand warmers used to elevate the mouse to the appropriate height and maintain body temperature. Inset shows the orientation of the Hamilton syringe in the automated injection apparatus.
Figure 2: Pictorial representation of the tooth placement on a stereotactic device, location of ear bars, and calvarial window relative to bregma. (A) The pictorial of the maxillary incisors in the tooth notch on the nose cone. (B) The location of the left and right ear bars within the medial canthus of the respective ears. (C) An arrow indicates the bregma and a red dot indicates the location where the calvarial window should be made (2 mm lateral to the sagittal suture and 1 mm anterior to the coronal suture).
The authors have nothing to disclose.
Surgical Materials | |||
Betadine | Purdue Products | 19-027132 | Povidone-iodine, 7.5% |
Bone Wax | Surgical Specialities | 903 | Sterile and malleable beeswax and isopropyl palmitate |
Buponorphine SR-Lab | ZooPharm | N/A | Long acting injectable analgesic 5 mL (0.5 mg/mL) polymetric formulation |
Cotton tip applicators | Puritan | 25-806 10WC | Sterile long stemmed cotton tip applicators |
Eye Ointment | Puralube | 17033-211-38 | Lubricating petrolatum and mineral oil based ophthalmic ointment |
Handwarmers | Hothands | HH2 | Air-activated heat packs |
Ibuprofen | Up & Up | 094-01-0245 | 100 mg per 5 mL in liquid suspension |
Isoflurane | Henry Schein INC | 1182097 | Liquid anesthetic for use in anesthetic vaporizer |
Scalpels | Integra Miltex | 4-410 | #10 disposable scalpel blade |
Suture | Covidien | SP5686G | 45 cm swedged 5-0 monofilament polypropylene suture |
Stereotaxic Unit | |||
High Speed Drill (Foredom) | Kopf | Model 1474 | Max of 38,000 RPM |
Mouse Gas Anesthesia Head Holder | Kopf | Model 923-B | Mouth bar with teeth hole and nosecone |
Non-Rupture Ear Bars | Kopf | Model 922 | Ear bars suitable for mouse applications |
Stereotaxic Instrument | Kopf | Model 940 | Base plate, frame and linear scale assembly with digital readout monitor |
Anesthesia Machine | |||
SomnoSuite Low-Flow Digital Vaporizer | Kent Scientific | SS-01 | Digital anesthesia machine |
SomnoSuite Starter Kit for mice | Kent Scientific | SOMNO-MSEKIT | Includes induction chamber, 2x anesthesia syringes, 18" tubing, plastic nosecone, 2x waste aneshesia gas canisters |