Focal cortical ischemia was induced by photothrombosis of cortical microvessels using Rose Bengal (Sigma-Aldrich, St. Louis, MO, USA) with cold light (Zeiss KL1500 LCD, Germany) 17 (link). Each animal was anesthetized with 5% isoflurane and anesthesia was maintained with 3% isoflurane in an oxygen/air mixture using a gas anesthesia mask in a stereotaxic frame (Stoelting, Wood Dale, IL, USA). Body temperature was maintained at 37 ± 0.5°C during surgery using a heating pad controlled by a rectal probe. For illumination, a 4.5 mm fiber-optic bundle from a cold light source was positioned on the exposed skull 0.5 mm anterior to the bregma and 3.7 mm lateral to the midline over the left sensorimotor cortex, as previously described 18 (link). The brain was illuminated for 10 minutes after infusion of 50 mg/kg Rose Bengal in normal saline into the right femoral vein via a microinjection pump within 1 minute. The scalp was sutured, and the mice were allowed to wake up before being returned to their home cages.
Gas anesthesia mask
Manufactured by Stoelting
Sourced in United States
The gas anesthesia mask is a medical device used to administer anesthetic gases to a patient during medical procedures. It is designed to cover the patient's nose and mouth to deliver a controlled mixture of gases, such as oxygen and anesthetic agents, to the patient's respiratory system.
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5 protocols using gas anesthesia mask
1
Photothrombotic Ischemic Stroke Model
2
Focal Cerebral Ischemia Induction in Mice
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Focal cerebral ischemia was induced by intraluminal middle cerebral artery occlusion (MCAO). Briefly, a 7/0 surgical nylon monofilament with rounded tip was introduced into the left internal carotid through the external carotid stump and advanced 10–13 mm past the carotid bifurcation. Occlusion was confirmed when blood flow was reduced by at least 80% of the baseline. The filament was left in place for 60 min and then withdrawn. The sham-operated animals were treated identically, except that the middle cerebral artery was not occluded after the neck incision. Each mouse was anesthetized with 2% isoflurane and maintained with 1% isoflurane in an oxygen/air mixture by using a gas anesthesia mask in a stereotaxic frame (Stoelting). The rectal temperature was maintained during surgery at 37 ± 0.5°C with a homeothermic blanket (Harvard Apparatus).
3
Photothrombosis-induced Focal Cortical Ischemia
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All surgical procedures and postoperative care were performed in accordance with the guidelines of the Chonnam National University Animal Care and Usage Committee, Korea. Sixty male Sprague-Dawley rats, weighing 230–300 g, were used. The animals were maintained on a 12 hour light/dark cycle and were allowed free access to food and water. The rats were anesthetized with 5% isoflurane and maintained with 2% isoflurane in an oxygen/air mixture using a gas anesthesia mask in a stereotaxic frame (Stoelting, Wood Dale, IL, USA). Focal cortical ischemia was induced by photothrombosis of the cortical microvessels using Rose Bengal (Sigma Chemical Co., St. Louis, MO, USA) with cold light (Zeiss KL1500 LCD, Oberkochen, Germany), as described previously (Watson et al., 1985). Measurements were made 1 hour, 12 hours, 1 day, 3 days, and 7 days after the onset of ischemia (n = 5 rats per group). The scalp was sutured and the rats were allowed to awaken before being returned to their home cages. Five animals received illumination after infusion of normal saline instead of Rose Bengal for the sham surgery.
4
Photothrombotic Focal Cortical Ischemia
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Focal cortical ischemia was induced by photothrombosis of the cortical microvessels using Rose Bengal (Sigma, St. Louis, MO, USA) with cold light (Zeiss KL1500 LCD, Germany) 5 (link),20 (link). Each animal was anesthetized with 5% isoflurane and maintained with 3% isoflurane in an oxygen/air mixture using a gas anesthesia mask in a stereotaxic frame (Stoelting, Wood Dale, IL, USA). Body temperature was maintained during surgery at 37 ± 0.5°C using a heating pad controlled by a rectal probe. For illumination, a 4.5 mm fiber-optic bundle from a cold light source was positioned onto the exposed skull 0.5 mm anterior to bregma and 3.7 mm lateral to the midline over the left sensorimotor cortex, as previously described 21 (link). The brain was illuminated for 10 minutes after infusion of 50 mg/kg Rose Bengal in normal saline into the right femoral vein via a microinjection pump within 1 minute. The scalp was sutured, and the mice were allowed to wake before being returned to their home cages. For the sham surgery, 6 animals received illumination after infusion of normal saline instead of Rose Bengal. Focal cerebral ischemia confirmation was also evaluated by using Evans blue (Sigma, St. Louis, MO, USA) dye extravasations via unaided examination or NIR fluorescence-based ring-enhancement.
5
Electroretinography Monitoring of Rat Visual Function
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Electroretinography (ERG) was performed every month after birth using the HMsERG system (Ocuscience, Las Vegas, NV) as previously described [8] (link). Briefly, after dark adapted overnight, the rats were anesthetized with an injection of ketamine/xylazine (37.5 mg/kg ketamine and 5 mg/kg xylazine, i.p.) and 0-2% isoflurane mixed with oxygen through a gas anesthesia mask (Stoelting, Wood Dale, IL, USA); and eyes were dilated using tropicamide 1% (Bausch & Lomb Inc., Tampa, FL) eye drops. Contact lens electrodes were placed on the cornea of both eyes, with reference and ground electrodes placed subcutaneously. An optically clear ophthalmic gel was used to maintain hydration and conductivity between the cornea and recording electrodes. Scotopic testing was conducted with flash stimuli intensities ranging from 1 to 25,000 millicandela (mcd) followed by photopic testing (light adaptation of 10 min prior to the photopic test which records flash stimuli responses of 10-25,000 mcd).
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