Medetomidine

AL was measured from the anterior corneal surface to the retinal pigment epithelium using a spectral domain optical coherence tomography (SD-OCT) system (Envisu R4310, Leica, Germany). The refraction was detected using an eccentric infrared photo refractor (Steinbeis Transfer Center, Germany) at the vertical pupil meridian. The data were automatically recorded by the program when the parameter values were stable. Choroidal thickness (ChT) was measured using SD-OCT, and the posterior surface of the choroid was quantified using Image J (Ver 1.53, NIH). ChT was determined using the formula: area divided by circumference. Intraocular pressure (IOP) was measured using a tonometer (Tono Lab, Icare Finland Oy, Vantaa, Finland) calibrated for mice under anaesthesia. Parts images of ocular components measurement were shown in Additional file 5: Fig. S5.
Mice were anesthetized with 0.75 mg/kg medetomidine (Sandoz K.K., Tokyo, Japan), 4 mg/kg midazolam (Domitor®, Orion Corporation, Espoo, Finland), and 5 mg/kg butorphanol tartrate (Meiji Seika Pharma Co., Ltd., Tokyo, Japan) dissolved in normal saline. Mydriasis was induced by 0.5% tropicamide eye drops (Santen, Osaka, Japan), and mice were placed in a cylindrical holder for measurement.

AL was measured from the anterior corneal surface to the retinal pigment epithelium using a spectral domain optical coherence tomography (SD-OCT) system (Envisu R4310, Leica, Germany). The refraction was detected using an eccentric infrared photo refractor (Steinbeis Transfer Center, Germany) at the vertical pupil meridian. The data were automatically recorded by the program when the parameter values were stable. Choroidal thickness (ChT) was measured using SD-OCT, and the posterior surface of the choroid was quantified using Image J (Ver 1.53, NIH). ChT was determined using the formula: area divided by circumference. Intraocular pressure (IOP) was measured using a tonometer (Tono Lab, Icare Finland Oy, Vantaa, Finland) calibrated for mice under anaesthesia. Parts images of ocular components measurement were shown in Additional file 5: Fig. S5.
Mice were anesthetized with 0.75 mg/kg medetomidine (Sandoz K.K., Tokyo, Japan), 4 mg/kg midazolam (Domitor®, Orion Corporation, Espoo, Finland), and 5 mg/kg butorphanol tartrate (Meiji Seika Pharma Co., Ltd., Tokyo, Japan) dissolved in normal saline. Mydriasis was induced by 0.5% tropicamide eye drops (Santen, Osaka, Japan), and mice were placed in a cylindrical holder for measurement.

Various murine retinal I/R techniques with different IOP and time have been addressed [33 (link),34 (link)], while in the current study, the method with IOP of 90–99 mmHg in 30 min was adopted. Mice were anesthetized using intraperitoneal injection of medetomidine (0.75 mg/kg, Sandoz K.K., Tokyo, Japan), midazolam (4 mg/kg, Domitor^®, Orion Corporation, Espoo, Finland) and butorphanol tartrate (5 mg/kg, Meiji Seika Pharma Co., Ltd., Japan) dissolved in normal saline (MMB). The mydriatic agent (Tropicamide, Phenylephrine Hydrochloride, Santen Pharmaceutical Co., Ltd., Osaka, Japan) eye drop was added to the eye. Cannulation of the anterior chamber was performed to increase IOP to 90–99 mmHg by infusion of normal saline into the eye with 35-gauge stainless needle. IOP was checked with a tonometer (TonoLab, Icare Finland Oy, Vantaa, Finland) every 5 min. The High IOP condition was maintained for 30 min, then the needle was pulled out.