Adult C57/BL6J mice (45–60 days old) were anesthetized with 80 mg/kg Ketamine, 8 mg/kg Xylazine and 1 mg/kg Acepromazine cocktail and then secured in the stereotaxic apparatus. Buprenorphine (1 mg/kg) was administered to induce analgesia. The scalp was shaved and cleaned with Iodine. A small midline incision of approximately 1 cm was made in the scalp to expose the dorsal surface of the skull. Tissue was dissected, and bone cleared with rubbing alcohol. Bregma and Lambda were set to the equal dorsoventral distance. Two bilateral injection sites were drilled through the calvaria with a micro drill bit at site 1: +/−1.8 lateral, −2.1 anteroposterior, 1.6 dorsoventral and site 2: +/− 2.5 lateral, −2.8 anteroposterior, 2.1 dorsoventral. These injection sites targeted the dentate gyrus of the dorsal hippocampus. A volume of 75 nl of AAV (4 × 1012 GC/ml) was injected at a rate of 7.5 nl/min. After confirming there was not bleeding at the injection site, the scalp was glued using Vetbond glue (3M™). Mice were place in a thermoregulated mat until recover from anesthesia.
Vetbond glue
Manufactured by 3M
Vetbond is a cyanoacrylate-based surgical adhesive developed by 3M for use in veterinary applications. It is designed to quickly and effectively bind tissue together, facilitating healing. The product is a sterile, single-use liquid adhesive that can be applied directly to the wound site.
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Lab products found in correlation
5 protocols using vetbond glue
1
Targeted AAV Injection in Murine Hippocampus
2
Stereotaxic Surgery and Craniotomy in Mice
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All procedures were carried out according to animal welfare guidelines authorized by the Baylor College of Medicine IACUC committee. All surgeries were performed under general anesthesia with 1.5% isoflurane. The mouse head was fixed in a stereotactical stage (Kopf Instruments), and eyes were protected with a thin layer of polydimethylsiloxane (30,000 cst, Sigma-Aldrich). After removing the scalp, a custom-made titanium headplate was attached to the skull with dental acrylic (Lang Dental). A 3 mm wide circular craniotomy centered 2.5 mm lateral of the midline and 1.2 mm anterior of the lambda suture was made, targeting the middle of the monocular region of left V1. A coverglass with a hole for pipette access was placed on the brain and carefully anchored with vetbond glue (3M, Saint Paul, MN) and dental acrylic (Lang Dental).
3
Visual Deprivation in Mouse Model
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Visual form deprivation (VFD) was induced in 12 mice by applying a unilateral frosted hemispherical plastic diffusers over the right eyes as previously described [60 (link)]. Briefly, frosted hemispherical plastic diffusers were hand-made using caps from 0.2 ml PCR tubes (Molecular BioProducts, San Diego, CA) and rings made from medical tape (inner diameter 6 mm; outer diameter 8 mm). A cap was frosted with fine sandpaper and attached to a ring with Loctite™ Super Glue (Henkel Consumer Adhesives, Avon, OH). On the first day of the experiment (P24), animals were anesthetized via intraperitoneal injection of ketamine (90 mg/kg) and xylazine (10 mg/kg), and diffusers were attached to the skin surrounding the right eye with several stitches using size 5–0 ETHILON™ microsurgical sutures (Ethicon, Somerville, NJ) and reinforced with Vetbond™ glue (3M Animal Care Products, St. Paul, MN). The contralateral untreated left eyes were used as control. Toenails were covered with adhesive tape to prevent mice from removing the diffusers. Animals recovered on a warming pad and were then housed in transparent plastic cages for the duration of the experiment (10 days). A control group comprised of 8 age-matched C57BL/6J mice was maintained under the same experimental conditions as experimental group, but without VFD.
4
Traumatic Brain Injury in Mice
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Mice were anesthetized by intraperitoneal MMF injection. Once the mice presented no reflex reaction from paw pinching, they were put on a stereotaxic frame (Precision Systems & Instrumentation, LLC). A skin incision was made followed by the drilling of a window on the right skull hemisphere, which was positioned rostrocaudal, between the bregma and lambda and under the sagittal suture. Mice were then placed on the TBI-0310 impactor (Precision Systems & Instrumentation, LLC) to undergo traumatic brain injury (TBI). The tip of a 3-mm diameter steel rod was used to induce injury to the somatosensory cortex according to the following settings: 6 m/s, 150 ms dwell time, 0.5 mm depth61 . Mice were removed from the impactor, the skull window was repositioned and sealed with Vetbond glue (3 M Vetbond, 3 M United States), and the skin was stitched. Mice were placed on a heating pad and injected with the antagonist mix before receiving a subcutaneous glucose injection (glucose 5% B. Braun Infusionslösung).
5
In vivo Imaging of Auditory Cortex
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Male and female H2B::PaGFP mice were used for in vivo imaging experiments and a small imaging window was implanted over the auditory cortex three weeks after virus injection. Briefly, mice were deeply anesthetized with a single dose of ketamine/medetomidine. After removal of an about 1 cm2 patch of skin over the parietal bones on the right lateral part of the head, a small part of the musculus temporalis was removed to expose the temporal bone. Using a dentist’s drill, the bones were smoothened, and part of the zygomatic process was removed and covered with a thin layer of Vetbond glue (3M). Next, a thin layer of dental cement (Lang Dental) was applied, except for the area over the temporal bone. The temporal bone was removed with a dental drill to expose a ~2 mm × 3 mm part of the brain containing the auditory cortex. The craniotomy was subsequently sealed with a drop of liquid agarose, a small round cover glass, and additional dental cement. In addition, a small titanium head post was implanted for fixation of the head during imaging.
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