AP-2β NCC KO mutants and their control littermates were weighed and anesthetized with an intraperitoneal (i.p.) injection of 2.5% avertin at 0.015 ml/g body weight. Whiskers were trimmed to ensure no obstruction of the imaging system and two sets of eye drops applied in order to dilate the pupils (0.5% Tropicamide Ophthalmic Solution and 2.5% Phenylephrine Hydrochloride Ophthalmic Solution, AKORN, Lake Forest, IL). The corneas were maintained well moistened throughout the procedure with consistent application of Tear-Gel ophthalmic liquid gel (Alcon Canada, Mississauga, ON, Canada) to prevent drying. The Phoenix Micron IV rodent eye imaging system, with optical coherence tomography (OCT) attachment (Phoenix Research Labs, Pleasanton, CA), was utilized to image the corneas and anterior chambers of the AP-2β NCC KO mutants and their control littermates.
Tropicamide ophthalmic solution
Manufactured by Akorn
Sourced in United States
Tropicamide ophthalmic solution is a pharmaceutical product used in ophthalmology. It is a mydriatic agent that dilates the pupil.
Automatically generated - may contain errors
Lab products found in correlation
Phenylephrine hydrochloride ophthalmic solution, by Akorn (9 mentions)
Metabond, by Parkell (7 mentions)
Custom coverglass, by Warner Instruments (6 mentions)
Celeris d430 rodent erg testing system, by Diagnosys (3 mentions)
Ophthalmic surgical microscope, by Zeiss (3 mentions)
Silflex 2, by World Precision Instruments (3 mentions)
Rpe kit, by World Precision Instruments (2 mentions)
Nf35bl 2, by World Precision Instruments (2 mentions)
Iol master, by Zeiss (2 mentions)
Phenylephrine hydrochloride ophthalmic solution, by Bausch & Lomb (2 mentions)
Gonak hypromellose ophthalmic demulcent solution, by Akorn (2 mentions)
Eye explorer, by Heidelberg Engineering (2 mentions)
Spectralis, by Heidelberg Engineering (2 mentions)
Genteal tears lubricant eye gel, by Alcon (2 mentions)
Microscope cover glass, by Thermo Fisher Scientific (1 mentions)
Genteal, by Novartis (1 mentions)
Rompun, by Bayer (1 mentions)
Genteal, by Alcon (1 mentions)
Spectralis oct, by Heidelberg Engineering (1 mentions)
Confocal scanning laser ophthalmoscope, by Heidelberg Engineering (1 mentions)
34 protocols using tropicamide ophthalmic solution
1
Corneal Imaging of AP-2β NCC Mutants
2
Electroretinography in Dark-Adapted Mice
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Mice were dark-adapted overnight before ERG recording. Under dim red light illumination, mice were anesthetized by intraperitoneal injection of a ketamine/xylazine mixture (87.5 mg/kg and 12.5 mg/kg, respectively), and pupils were dilated with 1% tropicamide ophthalmic solution (Akorn) for 2-3 minutes. Mice were placed on a Celeris D430 rodent ERG testing system (Diagnosys) with its heater on to maintain animals'
3
In Vivo Rat Eye Imaging Protocol
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We imaged both eyes of age-matched SD and LE rats. The experiments were conducted in agreement with the ARVO (the Association for Research of Vision and Ophthalmology) Statement for the Use of Animals in Ophthalmic and Vision Research and with the guidelines of the Florida International University’s Institutional Animal Care and Use Committee.
For imaging, an animal was anesthetized by intraperitoneal injection of ketamin (54 mg/kg body weight) and xylazine (6 mg/kg body weight). The eye to be imaged was treated with topical proparacaine hydrochloride ophthalmic solution (Akorn, 0.5%, USP) for topical anesthesia and tropicamide ophthalmic solution (Akorn, 0.5%, USP) for pupil dilation. A hard contact lens was put on the eye to prevent corneal dehydration and opacification. The sedated rat was restrained in an animal mount with five degrees of freedom.
For imaging, an animal was anesthetized by intraperitoneal injection of ketamin (54 mg/kg body weight) and xylazine (6 mg/kg body weight). The eye to be imaged was treated with topical proparacaine hydrochloride ophthalmic solution (Akorn, 0.5%, USP) for topical anesthesia and tropicamide ophthalmic solution (Akorn, 0.5%, USP) for pupil dilation. A hard contact lens was put on the eye to prevent corneal dehydration and opacification. The sedated rat was restrained in an animal mount with five degrees of freedom.
4
Subretinal Viral Vector Delivery in Mice
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Mice were anesthetized by intraperitoneal injection of a cocktail consisting of 20 mg/ml ketamine and 1.75 mg/ml xylazine in phosphate-buffered saline at a dose of 0.1–0.13 ml per 25 g body weight, and their pupils were dilated with topical administration of 1% tropicamide ophthalmic solution (Akorn, no. 17478-102-12). Subretinal injections were performed using an ophthalmic surgical microscope (Zeiss). An incision was made through the cornea adjacent to the limbus at the nasal side using a 26-gauge needle. A 34-gauge blunt-end needle (World Precision Instruments, no. NF35BL-2) connected to an RPE-KIT (World Precision Instruments, no. RPE-KIT) by SilFlex tubing (World Precision Instruments, no. SILFLEX-2) was inserted through the corneal incision while avoiding the lens and advanced through the retina. Each mouse was injected with 1 μl of viral vector prep per eye. Only mice that had more than 95% retinal coverage after subretinal injection and with minimal complications were kept for further evaluation.
5
Standardized Murine Retinal Imaging
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Imaging was performed in a laboratory room at ~ 400 lx. Stray light from electronic equipment was masked, and the imaging stage was shrouded with blackout fabric. Experiments were regularly performed between 12 and 3 pm. At the time of the experiment, the mouse retina was light-adapted due to constant ambient light conditions. The mouse was first anesthetized by intraperitoneal injection of a mixture of 100 mg/kg ketamine and 5 mg/kg xylazine. The anesthesia cocktail concentration was 10 mg/mL ketamine and 0.5 mg/mL xylazine. A drop of 1% tropicamide ophthalmic solution (Akorn, Lake Forest, IL) was applied to the imaging eye for pupil dilation, and a cover glass (12-545-80; Microscope cover glass, Fisherbrand, Waltham, MA) with a drop of eye gel (GenTeal, Novartis, Basel, Switzerland) was placed on the imaging eye. After the mouse was fully anesthetized, the head was fixed by a bite bar and ear bars in an animal holder. A heating pad was placed around the animal holder to keep the mouse warm. It took around 8–10 min between the time of intraperitoneal injection and the first OCT volume acquisition.
6
Measuring Intraocular Pressure and Axial Length
7
Rodent Electroretinography Protocol
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Mice were anesthetized by an intraperitoneal injection of ketamine (100 mg/kg, KetaVed, Bioniche Teoranta, Inverin Co, Galway, Ireland) and xylazine (10 mg/kg, Rompun, Bayer, Shawnee Mission, KS), and their pupils were dilated with 1% tropicamide (Tropicamide Ophthalmic Solution, Akorn, Lake Forest, IL). Thereafter, the corneas were moistened using 0.3% hypromellose gel (GenTeal, Alcon, Fort Worth, TX) which also secured electrical conductivity during electroretinography (ERG) recording. The ERG was performed as previously described [46 (link)] using a Diagnosys Celeris rodent ERG device (Diagnosys LLC, Lowell, MA). Briefly, mice were dark-adapted overnight, and all handling before ERG was done under dim red light (> 600 nm). After electrodes were attached, three more minutes were allowed to fully dark-adapt the animal before stimulation. Stimulation was performed using a green LED (peak emission at ∼ 544 nm, bandwidth ∼ 160 nm) and an ascending 1-log step light intensity series between 0.0005–50 cd s/m2.
8
Ferret Brain Imaging Surgical Protocol
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After 3-5 weeks of expression, ferrets were anesthetized with 12.5mg/kg ketamine and isoflurane. Atropine and bupivacaine were administered, animals were placed on a feedback-controlled heating pad to maintain an internal temperature of 37° Celsius, and intubated to be artificially respirated. Isoflurane was delivered throughout the surgical procedure to maintain a surgical plane of anesthesia. An intravenous cannula was placed to deliver fluids. Tidal CO2, external temperature, and internal temperature were continuously monitored. The scalp was retracted and a custom titanium headplate adhered to the skull (Metabond, Parkell). A craniotomy was performed and the dura retracted to reveal the cortex. One piece of custom coverglass (5 mm diameter, 0.7 mm thickness, Warner Instruments) attached to a custom insert using optical adhesive (71, Norland Products) was placed onto the brain to dampen biological motion during imaging.
A 1:1 mixture of tropicamide ophthalmic solution (Akorn) and phenylephrine hydrochloride ophthalmic solution (Akorn) was applied to both eyes to dilate the pupils and retract the nictitating membranes. Contact lenses were inserted to protect the eyes. Upon completion of the surgical procedure, isoflurane was gradually reduced and pancuronium (0.2 mg/kg/hr) was delivered IV.
A 1:1 mixture of tropicamide ophthalmic solution (Akorn) and phenylephrine hydrochloride ophthalmic solution (Akorn) was applied to both eyes to dilate the pupils and retract the nictitating membranes. Contact lenses were inserted to protect the eyes. Upon completion of the surgical procedure, isoflurane was gradually reduced and pancuronium (0.2 mg/kg/hr) was delivered IV.
9
Craniotomy and Cortical Imaging in Ferrets
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After 3-5 weeks of expression, ferrets were anesthetized with 50mg/kg ketamine and isoflurane. Atropine and bupivacaine were administered, animals were placed on a feedbackcontrolled heating pad to maintain an internal temperature of 37 degrees Celsius, and intubated to be artificially respirated. Isoflurane was delivered throughout the surgical procedure to maintain a surgical plane of anesthesia. An intravenous cannula was placed to deliver fluids. Tidal CO2, external temperature, and internal temperature were continuously monitored. The scalp was retracted and a custom titanium headplate adhered to the skull (Metabond, Parkell). A craniotomy was performed and the dura retracted to reveal the cortex. One piece of custom cover-glass (3mm diameter, 0.7mm thickness, Warner Instruments) adhered using optical adhesive (71, Norland Products) to custom insert was placed onto the brain to dampen biological motion during imaging.
A 1:1 mixture of tropicamide ophthalmic solution (Akorn) and phenylephrine hydrochloride ophthalmic solution (Akorn) was applied to both eyes to dilate the pupils and retract the nictating membranes. Contact lenses were inserted to protect the eyes. Upon completion of the surgical procedure, isoflurane was gradually reduced and pancuronium (2 mg/kg/hr) was delivered IV.
A 1:1 mixture of tropicamide ophthalmic solution (Akorn) and phenylephrine hydrochloride ophthalmic solution (Akorn) was applied to both eyes to dilate the pupils and retract the nictating membranes. Contact lenses were inserted to protect the eyes. Upon completion of the surgical procedure, isoflurane was gradually reduced and pancuronium (2 mg/kg/hr) was delivered IV.
10
Craniotomy and Cortical Imaging in Ferrets
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After 3-5 weeks of expression, ferrets were anesthetized with 50mg/kg ketamine and isoflurane. Atropine and bupivacaine were administered, animals were placed on a feedbackcontrolled heating pad to maintain an internal temperature of 37 degrees Celsius, and intubated to be artificially respirated. Isoflurane was delivered throughout the surgical procedure to maintain a surgical plane of anesthesia. An intravenous cannula was placed to deliver fluids. Tidal CO2, external temperature, and internal temperature were continuously monitored. The scalp was retracted and a custom titanium headplate adhered to the skull (Metabond, Parkell). A craniotomy was performed and the dura retracted to reveal the cortex. One piece of custom cover-glass (3mm diameter, 0.7mm thickness, Warner Instruments) adhered using optical adhesive (71, Norland Products) to custom insert was placed onto the brain to dampen biological motion during imaging.
A 1:1 mixture of tropicamide ophthalmic solution (Akorn) and phenylephrine hydrochloride ophthalmic solution (Akorn) was applied to both eyes to dilate the pupils and retract the nictating membranes. Contact lenses were inserted to protect the eyes. Upon completion of the surgical procedure, isoflurane was gradually reduced and pancuronium (2 mg/kg/hr) was delivered IV.
A 1:1 mixture of tropicamide ophthalmic solution (Akorn) and phenylephrine hydrochloride ophthalmic solution (Akorn) was applied to both eyes to dilate the pupils and retract the nictating membranes. Contact lenses were inserted to protect the eyes. Upon completion of the surgical procedure, isoflurane was gradually reduced and pancuronium (2 mg/kg/hr) was delivered IV.
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