The following surgical procedures were performed under deep Medetomidin (Domitor; Pfizer, New York City, NY, USA), Midazolam (Dormicum; Roche, Basel, Switzerland) and Fentanyl (Janssen Pharmaceutica, Beerese, Belgium) anesthesia (150 μg Medetomidin, 2 mg Midazolam, 5 μg Fentanyl per kg body weight i.m. initially and additional injections as needed). Animals were placed in a Kopf stereotaxic device (Kopf Instruments, Tujunga, CA, USA). Two small holes (1.5-2.0 mm diameter) were drilled in the skull at −3.8 mm from Bregma and 2.2 mm laterally at both hemispheres. A NanoFil syringe (World Precision Instruments, Inc., Sarasota, FL, USA) with a 35 gauge beveled needle (NF35BV-2; World Precision Instruments) was used to slowly inject 0.75 μl of the viral solution at 3.2 mm and 3.7 mm below the brain surface into the dentate gyrus (DG; both hemispheres).
Nf35bv 2
Manufactured by World Precision Instruments
Sourced in United States
The NF35BV-2 is a laboratory instrument designed for precision temperature control. It features a built-in temperature sensor and a programmable temperature control system to maintain a stable and accurate temperature environment.
Automatically generated - may contain errors
Lab products found in correlation
Nanofil syringe, by World Precision Instruments (2 mentions)
Medetomidin, by Pfizer (1 mentions)
Kopf stereotaxic device, by Kopf Instruments (1 mentions)
Midazolam, by Roche (1 mentions)
Dormicum, by Roche (1 mentions)
Fentanyl, by Johnson & Johnson (1 mentions)
Midazolam, by Johnson & Johnson (1 mentions)
Dfc7000 t camera, by Leica (1 mentions)
M205 fca microscope, by Leica (1 mentions)
N methyl d aspartate, by Merck Group (1 mentions)
Tropicamide, by Akorn (1 mentions)
Proparacaine hydrochloride, by Akorn (1 mentions)
4 protocols using nf35bv 2
1
Stereotaxic Viral Injection for Dentate Gyrus Transduction
2
Tracing Trigeminal Ganglion Innervation
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DiI crystals (Invitrogen, D282) were placed around individual whisker follicles of P0–P1 mice using a 35G beveled needle (World Precision Instruments, NF35BV-2), following which, the pups were warmed on a heating pad (37 °C) for 10–15 min before returning to the mother. Four to 5 days later, the pups were sampled, and the TG was carefully isolated while still attached to the cranial base. DiI labelling was observed in the TG and images were acquired using the M205 FCA microscope (Leica) and DFC7000T camera (Leica).
3
Viral Vector Therapy for Spinal Injury
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Injured animals were blindly separated into one of three groups (Control GFAP-mCherry, GFAP-Ascl1-mCherry and miR124+miR9/9*+NeuroD1-GFP) and decoded after all functional data analysis. Animals were anesthetized (xylazine-ketamine) as described above, the skin and underlying musculature were re-exposed one-week post-injury. Using a filling needle, 10μL of either control or active virus were preloaded into a 10μL Nanofil Syringe (World Precision Instruments, #NC0578818). The filling needle was replaced with a 35-gauge needle (World Precision Instruments, #NF35BV-2), and the syringe was secured to a micromanipulator (Model Kite-L, World Precision Instruments). Control (GFAP-mCherry, 107–108 pfu), Ascl1-mCherry (107–108 pfu), or miR-GFP (107–108 pfu), was injected into the contusion cavity (visualized by the bruise in the spinal tissue). The underlying muscle was sutured in layers and the skin was closed with wound clips as described above. Animals recovered for 8 weeks prior to terminal electrophysiology recordings.
4
Retinal Excitotoxicity Induction in Mice
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Retinal excitotoxicity was induced according to the method established in our laboratory [26 (link)]. Mice were anesthetized with 73 mg/kg ketamine hydrochloride and 7.3 mg/kg xylazine hydrochloride intraperitoneally. Pupils were dilated with 1% tropicamide (Akorn, Lake Forest, IL, USA). One drop of proparacaine hydrochloride (Akorn) was applied to the cornea as topical anesthesia. NMDA (N-Methyl-D-Aspartate, (Sigma, St. Louis, MO, USA), 20 n moles/eye, 1 µL, dissolved in saline) was injected intravitreally into the right eye using a beveled 35 G needle (NF35BV-2, World Precision Instruments, Sarasota, FL, USA) connected with an SGE Syringe (World Precision Instruments, Sarasota, FL, USA). The needle was moved out slowly after maintaining for 30 s. Antibiotic ointment was applied to prevent infection. NMLA (N-Methyl-L-Aspartate, 20 n moles/left eye) was used as control.
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