Mice were anesthetized via intraperitoneal (IP) delivery of ketamine (90 mg/kg; Akorn, Lake Forest, IL, USA)/xylazine (12 mg/kg; Akorn) cocktail. Eyes were anesthetized with 1 drop of 0.5% tetracaine (Alcon, Fort Worth, TX, USA). Pupils were dilated with 1 drop of 2.5% phenylephrine (Akorn) and 0.5% tropicamide (Akorn). The whiskers were trimmed with scissors. A subcutaneous injection of meloxicam (1 mg/kg; Henry Schein Animal Health, Melville, NY, USA) was given for pain control. Mice were moved to the slit lamp stage and a cover slip was coupled to the cornea using Goniosoft (OCuSOFT, Rosenberg, TX, USA) to allow direct retinal visualization. Mouse eyes were treated with 4 (CNV area quantitation) or 8 (flow cytometry) focal laser burns (75 um, 110 mW, 100 msec) in each eye using an IRIDEX (Mountain View, CA, USA) 532 nm argon ophthalmic laser delivered via a Zeiss (Oberkochen, Germany) slit lamp. IP injections of propranolol (20 mg/kg) or PBS were performed daily during the course of experiments.
Protocol detail
Laser-induced Choroidal Neovascularization in Mice
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Animal
Argon laser
Burns
Cornea
Delivery
Flow cytometry
Forest
Intraperitoneal injections
Ketamine
Meloxicam
Mouse
Pain control
Phenylephrine
Propranolol
Pupils
Retinal
Slit lamp
Subcutaneous injection
Tetracaine
Tropicamide
Whiskers
Xylazine
Corresponding Organization : Northwestern University
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Variable analysis
independent variables
- IP delivery of ketamine (90 mg/kg) and xylazine (12 mg/kg) cocktail
- IP injections of propranolol (20 mg/kg) or PBS
- Quantitation of CNV area
- Flow cytometry
- Anesthesia of eyes with 0.5% tetracaine
- Dilation of pupils with 2.5% phenylephrine and 0.5% tropicamide
- Trimming of whiskers
- Subcutaneous injection of meloxicam (1 mg/kg) for pain control
- Coupling of cover slip to the cornea using Goniosoft to allow direct retinal visualization
- Focal laser burns (75 um, 110 mW, 100 msec) using a 532 nm argon ophthalmic laser
- IP injections of PBS
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