mouse headpost implantation: a method for head fixation in rodents

Mouse Headpost Implantation: A Method for Head Fixation in Rodents

After confirming a lack of response to toe pinch in a sedated mouse, use a pair of blunt forceps to grasp the skin at the back of the head and use scissors to make a 1.5-centimeter longitudinal incision to expose the skull. Gently scratch the periosteum with a scalpel and apply a drop of green activator onto the middle of the skull to increase the bone permeability.
Next, mix one spoonful of polymer from the implantation kit with five drops of monomer and one drop of catalyzer, and brush a generous amount of cement mix between the lambda and bregma skull landmarks. Using a swiping motion from lambda to bregma, quickly place the headpost onto the cement, and reapply additional cement around the inferior part of the post to ensure that the headpost sticks securely to the skull.
While the cement is drying, mix the resin powder with the resin liquid to a smooth consistency and apply the resulting resin mixture to the dried cement and around the headpost. After allowing the resin to dry for 3 minutes, use a vicryl 4-0 suture to close the skin at the back of the ears and use a cotton swab to apply a 10% to 20% iodine solution to the incision. Then, place the animal under a red light with monitoring until full recovery.

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All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Headpost Implantation Surgery
NOTE: Ensure the use of sterile instruments and arrange surgery and recovery in different zones. Once mastered, the implantation procedure lasts about 30 min.
<ol>
	<li>For analgesia, 30 min before the beginning of the surgery, subcutaneously inject buprenorphine (0.05 mg/kg) and put back the animal in its home cage. 
	NOTE: Buprenorphine&#39;s analgesic effects last approximately 12 h, long after the end of the procedure. In our experience, mice do not show any signs of distress related to this intervention&#160;but a subsequent dose of 0.05 mg/kg buprenorphine is recommended 24h after the surgery.&#160;</li>
	<li>Anesthetize the animal in a chamber with 2.5%&#8211;3% isoflurane gas. Wait 3 min and check if the mouse is properly anesthetized by observing respiration and lack of movement inside the chamber. Pass the mouse to a nose cone on a surgical table with a heating pad and, by interdigital pinching, verify that there is no withdrawal reflex and lower the isoflurane to 1.5%.</li>
	<li>Shave the head of the mouse using an electric razor. To obtain a sterile environment, rub the shaved area with iodine solution and after with 70% alcohol. Repeat this procedure two more times.</li>
	<li>Inject lidocaine hydrochloride (2%, 2 mg/kg) under the skin of the head for local anesthesia and wait 5 min for the effects to begin. To avoid eye damage due to dryness, cover the mouse&#39;s eyes with topical ophthalmic vet ointment.</li>
	<li>With a pair of blunt forceps, grab the skin at the back of the head, and with a pair of blunt scissors (or scalpel), make a longitudinal incision of about 1.5 cm to expose the skull.</li>
	<li>With the help of a scalpel, scratch the periosteum. Be careful not to scratch too hard, as the fixation of the headpost can be compromised if the skull starts to bleed slightly.</li>
	<li>Apply a drop of the green activator (see&#160;Table of Materials) on the middle of the skull. This will improve the fixation of the cement by increasing bone permeability.</li>
	<li>Prepare the cement: mix one spoon (see&#160;Table of Materials) of polymer with five drops of monomer and one drop of catalyzer. With the help of a brush, apply a generous amount of the cement mix between the lambda and bregma skull landmarks.</li>
	<li>Quickly place the headpost on the cement with a swiping motion going from lambda to bregma. After the headpost has been placed, reapply more cement around the inferior part to ensure that the headpost properly sticks to the skull. To guarantee proper fixation, make sure the cement is applied abundantly and that it dries before continuing to the next step. 
	NOTE: With this fixation procedure, the headpost will not come off and allow for long-term, repeated tests; in our hands, headpost removal is &#60;10%.</li>
	<li>Prepare the resin mix by applying a powder-to-liquid ratio that enables a smooth consistency of the mixture. Apply the resin where the cement was applied as well as around the headpost in order to protect its surface.</li>
	<li>Wait 3 min for the resin to dry and close the skin at the back of the ears with monofilament suture. With a cotton swab, apply diluted (10%&#8211;20%) iodine solution to the operated area. 
	NOTE: Make sure the skin does not get stuck to the resin.</li>
	<li>Turn off the anesthesia and place the animal under a red warm light to avoid hypothermia. Place moistened food and&#160;hydrogel or another water source based in gel in the cage&#39;s floor. Do not leave the mouse unattended until it regains consciousness. As soon as the animal fully recovers from the procedure (usually, 30 min to 1 h after), place it in a cage with groups of three or four to stimulate social interactions.</li>
</ol>

<table><tbody><tr><td>Blunt scissors</td><td>FST</td><td>14079-10</td><td></td></tr><tr><td>Catalyst V</td><td>Sun Medical, Japan</td><td>LX22</td><td>Parkell bio-materials, Kit n&amp;deg;S380</td></tr><tr><td>Dentalon Plus</td><td>Heraeus</td><td>37041</td><td></td></tr><tr><td>Green activator</td><td>Sun Medical, Japan</td><td>VE-1</td><td>Parkell bio-materials, Kit n&amp;deg;S380</td></tr><tr><td>Monomer</td><td>Sun Medical, Japan</td><td>MF-1</td><td>Parkell bio-materials, Kit n&amp;deg;S380</td></tr><tr><td>Ocrygel</td><td>TvmLab</td><td>10779</td><td>Ophtalmic vet ointment</td></tr><tr><td>Polymer L-type clear (cement)</td><td>Sun Medical, Japan</td><td>TT12F</td><td>Parkell bio-materials, Kit n&amp;deg;S380</td></tr></tbody></table>

Source: Fran&#231;a de Barros, F., et al. <a href="https://www.jove.com/video/59135/" target="_blank">Long-term Sensory Conflict in Freely Behaving Mice</a>. J. Vis. Exp. (2019).
This video shows a headpost implantation surgery in murine model. The headpost is used not only to head-fix and therefore restrain the mouse during experimentation, but also to securely attach a head device in freely moving animals.

Source: Fran&#231;a de Barros, F., et al. Long-term Sensory Conflict in Freely Behaving Mice. J. Vis. Exp. (2019).
This video shows a headpost ...

On an anesthetized mouse whose head is shaved and prepared for surgery, make a length-wise incision to expose the skull. Next, gently remove the periosteum membrane to expose the surface of the skull. This will help the cement adhere better to the bone.
Apply activator to increase bone permeability. Then, in the area between the lambda and bregma landmarks of the skull, apply freshly mixed cement. Slide the headpost from lambda to bregma and apply more cement to ensure it is securely fixed.
Once the cement is completely dry, apply freshly mixed resin on top of the cement and around the headpost. Once the resin is dry, close the cut by suturing the skin.

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Mouse Headpost Implantation: A Method for Head Fixation in Rodents

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Mouse Headpost Implantation: A Method for Head Fixation in Rodents