The mice were anesthetized with Zoletil (I.M.) and restrained in a stereotactic apparatus (Stoelting Company, Wood Dale, Illinois, USA). A midline sagittal incision was made in the scalp. Okadaic acid (100 ng/1 μL in ACSF (147 mM NaCl, 2.9 mM KCl, 1.6 mM MgCl2, 1.7 mM CaCl2, and 2.2 mM dextrose) or only ACSF (1 μL) was ICV administered once into left lateral cerebral ventricle by microsyringe (Model 1701N, needle size 26s gauge) (Hamilton Company, NV, USA), using the coordinates: 0 mm posterior; −2.0 mm lateral; −2.5 mm ventral to bregma. After microsyringe puncture into cerebral ventricle and waiting for 5 min to obtain the equilibrium of brain pressure, microinjection continues for a period of 1 min (1 μL/min) by syringe pump (KD Scientific Inc., Holliston, MA, USA) then followed by an additional 5 min waiting time to allow for diffusion away from the injection site. Finally, the wound would be sutured rapidly.
Stereotactic apparatus
Manufactured by Stoelting
Sourced in United States, Germany
The Stereotactic Apparatus is a laboratory instrument used to precisely position and immobilize a subject, such as an animal, during experimental procedures. It provides a stable and reproducible frame of reference for targeting specific anatomical structures within the subject.
Automatically generated - may contain errors
Lab products found in correlation
Sprague dawley rats, by BioLASCO (4 mentions)
6 ohda, by Merck Group (3 mentions)
Hamilton syringe, by Stoelting (2 mentions)
Xylavet, by CP-Pharma (2 mentions)
Zoletil 50, by Virbac (2 mentions)
Axio imager, by Zeiss (2 mentions)
Syringe pump, by KD Scientific (1 mentions)
Axio scope a1, by Zeiss (1 mentions)
Axiocam 305 color, by Zeiss (1 mentions)
Fluorescent microscope, by Leica (1 mentions)
Imagepro version 6, by Media Cybernetics (1 mentions)
Matrigel, by Corning (1 mentions)
Metacam, by Boehringer Ingelheim (1 mentions)
Microinfusion pump, by Stoelting (1 mentions)
Female c57bl 6 mice, by Jackson ImmunoResearch (1 mentions)
C57bl 6j female mice, by Jackson ImmunoResearch (1 mentions)
Stainless steel electrode, by A-M Systems (1 mentions)
6 hydroxydopamine hydrochloride, by Merck Group (1 mentions)
C6 glioma cells, by American Type Culture Collection (1 mentions)
Ascorbic acid, by Merck Group (1 mentions)
41 protocols using stereotactic apparatus
1
Okadaic Acid Intracerebroventricular Injection
2
Intracerebral Hemorrhage Rat Model
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Fifty four male Sprague–Dawley rats (n = 9), weighing between 300–400 g (purchased from the BioLasco Taiwan Co., Ltd., authorized by Charles River Lab), were used in this study. All experimental protocols were approved and supervised by the University of Kaohsiung Medicine Animal Research Committee and in accordance with the Declaration of Helsinki (1964). The rats received anesthesia by an intraperitoneal injection of a mixture of 0.9 mg/100 gm xylazine and 5.5 mg/100 gm KetaVed. 1 ml/kg body weight (BW) fresh arterial blood was withdrawn from tail artery and injected into the craniocervical junction using a stereotactic apparatus (Stoelting, Wood Dale, IL 60191, USA) [45 (link)]. No mortality was found during the study. After the induction, animals were placed in ventral recumbent position for 30 minutes to let ventral blood clot formation. The repeated induction was performed 48 hr after the 1st induction. After monitoring for respiratory distress and giving mechanical ventilation if necessary, the animals were returned to the vivarium till fully awake. A habitat was offered with a 12 h light–dark cycle and an access to food and water ad lib.
3
Retrograde Labeling of Visual Pathways
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Deeply anesthetized rats (see above; n=6 in each group) were placed in a stereotactic apparatus (Stoelting, Kiel, Germany), and the skin overlying the skull was cut open. The lambda and bregma sutures served as landmarks for drilling four holes. FG (Biotium, Hayward, CA) was injected (8 μl of 4% FG in distilled H2O) in the superior colliculus and lateral geniculate bodies [20 (link)-22 (link)]. To allow for the retrograde transport of FG, the animals were kept for 6 days before further experimentation.
4
Retrograde Labeling of Retinal Ganglion Cells
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These procedures have been further explained in our previous studies [34 (link),35 (link)]. At day 21 after rAION induction, rats were anesthetized and placed in a stereotactic apparatus (Stoelting, Wood Dale, IL, USA). The skull was exposed, and 2 μL of 5% FG was injected into the superior colliculus on each side via a Hamilton syringe. After one week, the eyeballs were obtained after the rats had been euthanized. The eyeballs were incubated in 10% formalin for 2 h. Each retina was dissected and flat-mounted on a slide. The morphometry of the RGCs was detected by fluorescence microscopy (Axio Scope A1, Zeiss, Oberkochen, Germany) with a filter set (excitation filter = 350–400 nm, emission filter = 515 nm) and photographed with a digital camera (Axiocam 305 color, Zeiss, Oberkochen, Germany). The number of RGCs at a distance of 1 or 3 mm from the ONH was counted for determination of the central and mid-peripheral RGC densities. Eight random regions per slide were imaged in both the central and mid-peripheral areas. RGC density was calculated by ImageJ software.
5
Subcutaneous and Deep Brain Electrode Implantation for ABR and DBS
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Subcutaneous electrodes were implanted for ABR recordings and during the same surgery DBS electrodes were implanted in the brain (Fig. 1 ). Animals were anesthetized by intraperitoneal administration of ketamine (90 mg/kg) and xylazine (10 mg/kg). The head of the rats was immobilized in a stereotactic apparatus (Stoelting Co, Wood Dale, IL, USA) with mouth and blunt ear-bars. Permanent Teflon-coated stainless steel electrodes were subcutaneously implanted. One wire electrode was subcutaneously tunneled to the mastoid and a second wire electrode was attached to a screw on the vertex. Based on coordinates from a stereotactic atlas (Paxinos & Watson, 2007 ), bilateral electrodes (Technomed, Beek, The Netherlands) were inserted in the CIC (bregma −8.8, depth 4.5, interspace 3.8) or in the DCBN (bregma −11.5, depth 6.5, interspace 6.8). The postoperative recovery time was one week.
6
In vivo Retrograde Labeling of Retinal Ganglion Cells
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To assess the effect of DHPG on RGCs in vivo, retrograde labeling of RGCs was performed. After anesthetization, the rats were placed in the stereotactic apparatus (Stoelting, Wood Dale, IL, USA) and the brain surface was exposed by perforating the parietal bone to facilitate dye injection. 2 μL of 2% fluorogold (FG; Biotium, Hayward, CA, USA) was injected into both superior colliculi and dorsal lateral geniculate nuclei. After seven days, rats from different groups were sacrificed and eyeballs were enucleated and placed in 4% paraformaldehyde for 4 h. The whole retina was then carefully dissected, flattened, and mounted with the vitreous side up on slides. Photographs were captured using a fluorescent microscope (Leica, Germany) and FG-labeled RGCs were counted in a masked fashion by the same investigator using automated particle counting software in ImagePro Version 6.0 (Media Cybernetics, Bethesda, USA). The number of labeled cells in 12 photographs of each retina (three photographs per retinal quadrant) at 1/6, 3/6, and 5/6 of the retinal radius was summed together and expressed as mean RGC densities/mm2 for each group.
7
Intracranial Tumor Induction in Rats
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Intracranial tumors were established by stereotactic implantation of DIPG007 cells in the brain of 4-week-old RH-Foxn1rnu rats (n = 8, 102 ± 8 g). Rats were anesthetized with isoflurane (3% for induction and 2% for maintenance) in O2. 0.05 mg.kg−1 of buprenorphine was subcutaneously administered at the end of the intervention to prolong analgesia. 7.5 × 105 DIPG007 cells dispersed in 5 µL (70% Matrigel® (Corning, Corning, NY, USA), 30% DMEM F12/Neurobasal A 1:1) were injected using a Hamilton syringe in a stereotactic apparatus (Stoelting, Wood Dale, IL, USA) into the IVth ventricle. Stereotactic coordinates were as follows: 1 mm posterior and 2 mm lateral from the lambda, and 8 mm below the dura. A sham group of RH-Foxn1rnu rats (n = 4; 81 ± 8 g) were injected with 5 µL of the cell injection medium only at the exact coordinates.
8
Retrograde Labeling of Retinal Ganglion Cells
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Wild-type adult male mice were anesthetized and placed in a stereotactic apparatus (Stoelting, Kiel, WI, United States) as we did before (Lin et al., 2012 (link)). The skull was exposed and cleaned with 3% hydrogen peroxide. The accurate location of superior colliculus was conducted followed mice brain atlas (The Mouse Brain in Stereotaxic Coordinates, Academic Press, New York, 2001). A hole of 1 mm in diameter was drilled in the skull (4.4 mm posterior and 0.06 mm lateral to the bregma), and a 26-gauge stainless steel cannula was inserted for infusion of fluorochrome, hydroxystilbamidine (FG; Biotium, Inc., Hayward, CA, United States) at a speed of 1 μl/10 min. One week before ON lesion. One microliter of 2% FG was injected into the bilateral superior colliculus (1.2 mm deep from the skull). To analyze the survival number of RGCs in whole retinas labeled with FG at 7-d post-ON crush (ONC7d), the golden dots (survived RGCs) were counted using FIJI software (NIH, United States).
9
Intracerebral Hemorrhage Induction in Rats
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Forty-five male Sprague-Dawley rats, weighing between 350–450 g (BioLasco Taiwan Co., Ltd., authorized by Charles River Lab), were used in this study. All experimental protocols were approved and under supervision by the University of Kaohsiung Medicine Animal Research Committee. The rats were anesthetized by an intramuscular injection of a mixture of 0.9 mg/100 gm xylazine and 5.5 mg/100 gm KetaVed. 0.3 mL arterial blood was withdrawn and injected into the cisterna magna using a 30-gauge needle under a stereotactic apparatus (Stoelting, Wood Dale, IL, US). After the induction, animals were placed in ventral recumbent position for 15 minutes to allow ventral blood clot formation. Perfusion-fixation on the animals was then performed. After close monitoring for respiratory distress and giving mechanical ventilation if necessary, the animals were returned to the vivarium when fully awake. They were placed in a habitat with a 12 hr light-dark cycle and had access to food and water ad libitum.
10
Glycyrrhizin Pretreatment in Subarachnoid Hemorrhage
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The rats were randomly divided into the following five groups (nine animals/group): (1) sham operated (no SAH); (2) SAH only; (3) SAH plus vehicle; (4) SAH plus glycyrrhizin (5 mg/kg/day); and (5) SAH plus precondition glycyrrhizin (5 mg/kg/day). The dosage was adjusted according to our pilot study. The first injections given to animals were administered 24 hr before (pretreatment) and 1 hr after induction of SAH (treatment) by using an osmotic mini-pump (Alzet corp, Palo Alto, Ca). After the animals received reanesthesia, CSF sampling was obtained through a 30-gauge needle into the cranial-cervical junction by using stereotactic apparatus (Stoelting, Wood Dale, IL, US). The animals were sacrificed by perfusion-fixation 72 hr after 2nd SAH. Cortical tissue samples were obtained by means of placing a 22-gauge needle inserted 5 mm in depth into the skull bone (N = 5) through a burr hole craniectomy (2 mm apart from the bregma) at 24, 48, and 72 hrs.
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