The rats were anesthetized with tiletamine hydrochloride (60 µg/g) and fixed in a prone position using a Model 900 Small Animal Stereotaxic Instrument (David Kopf Instruments, Tujunga, CA, USA). Following exposure of the skull, a metallic disc of 20 mm in diameter and 2 mm in thickness was placed on the bregma. Using a device designed by Marmarou et al. [8 (link)] and Foda and Marmarou [9 (link)], we inserted a tube catheter of 120 cm in length and 22 mm in diameter. Then, we made two holes in the tube catheter at a gap distance of 5 cm to minimize air resistance. As proposed by Ucar et al. [10 (link)], we dropped an object of 450 g in weight onto the rat's head from a height of 1 m, inducing diffuse TBI. A total of 51 rats were used for the experimental procedure and 21 died of skull fracture and subsequent bleeding.
Model 900 small animal stereotaxic instrument
Manufactured by Kopf Instruments
Sourced in United States
The Model 900 Small Animal Stereotaxic Instrument is a precision laboratory device designed for the study of small animal brains. It provides a stable platform to securely hold the animal's head in a fixed position, enabling accurate and reproducible placement of electrodes, injections, or other interventions within the brain.
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Lab products found in correlation
Homeothermic blanket system, by Harvard Apparatus (1 mentions)
Trypsin edta, by MP Biomedicals (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
4 protocols using model 900 small animal stereotaxic instrument
1
Diffuse Traumatic Brain Injury Rat Model
2
Rat Model of Traumatic Brain Injury
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Rats were anesthetized through an intramuscular injection of tiletamine hydrochloride (60 mg/kg) into the thigh muscle and fixed to the prone position with a stereotaxic instrument (Model 900 Small Animal Stereotaxic Instrument; David Kopf Instruments, Tujunga, CA, USA). A Homeothermic pad (Homeothermic Blanket System; Harvard Apparatus Ltd., Edenbridge, Kent, UK) maintained the body temperature at 37.0°C±0.5°C. For inducing the brain injury, we applied the weight drop model instrument modified by Feeney et al. [10 (link)], adapting the method of Ducker [11 ] to induce a spinal cord injury. The protocol we followed was suggested by Yoon et al. [12 (link)] for inducing a moderate brain injury in rats (Fig. 1 ). The impact point was 1.0 mm anterior and 3.0 mm lateral to the bregma, which was the contralateral side of the dominant forelimb referred to on a rat brain map [13 ]. Above the impact point, an incision of 2.5 cm on the scalp was cut and after a circular craniotomy (d=3 mm) the dura mater was dissected and the cortical impact lesion was induced by dropping a round piece of brass with cross-sectional diameter of 2 mm onto the rat’s brain.
3
Establishing Rat C6 Glioma Xenograft Model
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The rat C6 glioma cell line has been used in previous studies as it exhibits similar characteristics to human gliblastoma cells, such as invasive growth and atypical nuclei (16 (link),17 (link)). The C6 glioma line was provided by the School of Biomedicine, Far Eastern Federal University for the experiment. An aliquot containing 1×106 tumor cells was thawed for 5 min at 37°C and washed free of dimethyl sulfoxide (Sigma-Aldrich, St. Louis, MO, USA), using Dulbecco’s modified Eagle’s medium (DMEM; Gibco Life Technologies, Carlsbad, CA, USA) containing 10% fetal bovine serum (FBS; Life Technologies) and 100X antibiotic-antimycotic (10,000 U/ml; Life Technologies). The cells were precipitated by centrifugation (120 × g for 3 min), fresh medium (DMEM) was added and then the cells were seeded into 50-ml culture flasks. Cultivation was continued until monolayer formation. Next, the cells were detached by enzymatic dissociation (0.05% trypsin-EDTA; MP Biomedicals, Santa Ana, CA, USA; 1:4; 10 min; 37°C), and after centrifugation (120 × g for 3 min) the supernatant was discarded and the cells were resuspended in fresh medium. Tumorigenicity was analyzed following the implantation of 0.5×106 C6 glioma cells into the brain of adult rats using a stereotaxic device (Model 900 Small Animal Stereotaxic Instrument; David Kopf Instruments, Tujunga, CA, USA) as described previously (5 (link)).
4
GL261 Glioblastoma Mouse Model
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The GL261 mouse glioma cell line was obtained from the National Cancer Institute Tumour Repository, Frederick, MD, USA. For stereotactic implantation of the cells, the Tspo+/+ (n = 4) and Tspo−/− mice (n = 12) were operated under isoflurane anaesthesia using a Model 900 Small Animal Stereotaxic Instrument and a Model 1911 Stereotaxic Drill (David Kopf Instruments, Tujunga, CA, USA). A small burr hole was made 3 mm anterior to the bregma and 2 mm lateral over the right hemisphere. GL261 glioblastoma cells were injected slowly to a depth of 3 mm ventral at 3 ×103 per µl in a volume of 5 µl over 2 min using a 10-µl Hamilton syringe connected to a pump by means of a 32-gauge needle. The animals were monitored daily for good recovery and absence of neurological signs. Three and four weeks after the operation, the animals were imaged by means of microPET/CT and the brains were dissected for histological studies and autoradiography.
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