In order to measure the average distance from RNFL to ONL, PS-OCT (Heidelberg, Germany) were performed for imaging in vivo. After intraperitoneal injected with chloral hydrate (5%, 0.075 ml/g) for deep anesthesia, pupils were dilated with compound tropicamide eye drop (Santen, Japan), using the circular scanning mode with the diameter of 3.45 mm (system setting) and centering on optic disc to get images through a 25D lens (50744, Heidelberg, Germany). The data was computed with ImageJ software.
Compound tropicamide eye drop
Manufactured by Santen
Sourced in Japan
Compound tropicamide eye drops are a type of ophthalmic solution containing the active ingredient tropicamide. Tropicamide is a parasympatholytic agent that is used to dilate the pupil of the eye.
Automatically generated - may contain errors
Lab products found in correlation
Oxybuprocaine hydrochloride eye drop, by Santen (2 mentions)
Pentobarbital sodium, by Merck Group (2 mentions)
Oxybuprocaine hydrochloride, by Santen (2 mentions)
Levofloxacin eye drops, by Santen (1 mentions)
Prednisolone acetate ophthalmic suspension, by Abbvie (1 mentions)
Sodium hyaluronate eye drops, by Santen (1 mentions)
Proparacaine hydrochloride eye drops, by Alcon (1 mentions)
Surgical microscope, by Zeiss (1 mentions)
Kr8800, by Topcon (1 mentions)
Ketamine, by Pfizer (1 mentions)
Kr 8100, by Topcon (1 mentions)
Iolmaster 500, by Zeiss (1 mentions)
12 protocols using compound tropicamide eye drop
1
In vivo RNFL to ONL Distance
2
Small Animal Refractive Measurements
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Refractive measurements for both eyes of all animals were taken at 0W, after 2 weeks (2W), and after 4 weeks (4W) using a small animal photorefractor (SriaTech Company, Germany) set at the following parameters: wavelength: 875 nm; frame rate: > 115 Hz; working distance: 56 cm. Before each measurement, pupils were dilated using a compound tropicamide eye drop (Santen Pharmaceutical Co., Ltd., Japan) for 10 min. All measurements were conducted in a dark room, and the average value of the refractive error was recorded and saved once it stabilized.
3
Implantation of V4c Implantable Collamer Lens
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All V4c ICL implantation surgeries were performed by the same ophthalmic surgeon. Compound tropicamide eye drops (1 ml: 5 mg: 5 mg, Santen, Japan) were applied 30 min before surgery to dilate the PD to 7 ± 0.5 mm. After the application of oxybuprocaine hydrochloride eye drops (20 ml: 80 mg, Santen, Japan) for topical anesthesia, a 3 mm temporal corneal incision and an auxiliary incision at 12 o'clock were made. The anterior chamber was injected with sodium hyaluronate (Shanghai Kinsson, China), a viscoelastic agent (OVD). The V4c ICL was inserted into the anterior chamber with a MicroSTAAR injector (STAAR Surgical Co., Monrovia, CA, USA) and then placed in the posterior chamber and positioned in the horizontal axis. OVD was exchanged with balanced salt solution. A 0.1 ml carbachol injection (2 ml: 0.1 mg, Freda, China), as the miotic agent, was instilled in the anterior chamber of the left eyes. After surgery, prednisolone acetate ophthalmic suspension (5 ml: 50 mg, Allergan, Ireland) and levofloxacin eye drops (5 ml: 24.4 mg, Santen) were administered topically four times daily for 2 weeks. No more mydriatic drops were applied between the procedure and each follow-up visit.
4
Electroretinography Protocol for Mouse Vision
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Electroretinograms were recorded 4 weeks after intravitreal injection, in accordance with the International Society for Clinical Electrophysiology of Vision. The recording electrode was a ring-shaped corneal contact silver-plated copper electrode with a diameter of 3 mm, and the reference electrode and ground electrode were self-made needle electrodes. Mice were placed in absolute darkness for >12 hours for dark adaptation. The mice were anesthetized and administered compound tropicamide eye drops (Santen Pharmaceutical, Chongqing, China). The head and limbs of the mouse were fixed on the detection platform in the dark room, the recording electrode was placed on the cornea, the reference electrode pierced under the skin of the mouse's front cheek, and the ground electrode pierced under the skin of the mastoid.
This experiment used the Roland visual electrophysiological system (Roland Consult, Brandenburg, Germany), and the stimulator was a Roland Ganzfeld Q 450. The maximum mixed-response stimulation frequency was 0.095 Hz, and the recording duration was 150 ms. Maximum mixed-reaction a- and b-wave amplitudes were recorded and analyzed. The a-wave amplitude refers to the peak from baseline to the a-wave bottom, and the b-wave amplitude refers to the peak from the a-wave bottom to the b-wave. The entire recording process was carried out under a weak red light.
This experiment used the Roland visual electrophysiological system (Roland Consult, Brandenburg, Germany), and the stimulator was a Roland Ganzfeld Q 450. The maximum mixed-response stimulation frequency was 0.095 Hz, and the recording duration was 150 ms. Maximum mixed-reaction a- and b-wave amplitudes were recorded and analyzed. The a-wave amplitude refers to the peak from baseline to the a-wave bottom, and the b-wave amplitude refers to the peak from the a-wave bottom to the b-wave. The entire recording process was carried out under a weak red light.
5
Rabbit Electroretinogram Recording Protocol
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After general anesthesia, compound tropicamide eye drops (Santen Pharmaceutical Co., LTD, Shiga Plant, Japan) were instilled in rabbits’ eyes to dilute the pupil, and 0.5% proparacaine hydrochloride eye drops (Alcon, Belgium) were used as corneal surface anesthesia. The circular corneal electrode was placed on the surface of the cornea of the rabbit, and the reference electrode of the silver needle was placed subcutaneously near the eye socket. The ground electrode of the silver needle was inserted subcutaneously into the back of the rabbit’s ear. ERG signals were amplified by an amplifier (Brownlee Precision Model 410, United States) at 128 Hz, and bandpass filtered between 1 and 1000 Hz. Light stimuli was applied by white LED and controlled by self-written Arduino code. Each session contains 10 stimuli, which lasts for 200 ms and is separated by 10 to 15 s randomly. The rabbit’s cornea was lubricated with 0.3% sodium hyaluronate eye drops (Santen Pharmaceutical Co., LTD, Shiga Plant, Japan) during recordings.
6
Intravitreal Injection and Optic Nerve Crush
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Intravitreal injection and ONC were performed as previously described (Huang et al., 2019b). Briefly, after anesthesia by intraperitoneal injection of 1% sodium pentobarbital (8 mL/kg; Sigma, St. Louis, MO, USA), the left eye was first dilated with compound tropicamide eye drops (Santen Pharmaceutical Co., Ltd., Osaka, Japan) and then 1 µL of AAV (approximately 2.5–3.0 × 1012 genome copies) was injected into the vitreous chamber using a 30G Hamilton syringe (Hamilton Co., Bonaduz, Switzerland).
Two weeks after injection, the left optic nerve was exposed intraorbitally under a surgical microscope (Carl Zeiss, Oberkochen, Germany) and crushed for 3 seconds with cross-action forceps 1 mm behind the eyeball to avoid disrupting the blood supply to the optic nerve. For the sham operation, the optic nerve was exposed but not crushed. For the selection of the crushing time, we referred to previous literature (Watkins et al., 2013; Miao et al., 2016; Tran et al., 2019). After surgery, the mouse was placed on a heating pad and monitored until it had fully recovered from anesthesia.
Two weeks after injection, the left optic nerve was exposed intraorbitally under a surgical microscope (Carl Zeiss, Oberkochen, Germany) and crushed for 3 seconds with cross-action forceps 1 mm behind the eyeball to avoid disrupting the blood supply to the optic nerve. For the sham operation, the optic nerve was exposed but not crushed. For the selection of the crushing time, we referred to previous literature (Watkins et al., 2013; Miao et al., 2016; Tran et al., 2019). After surgery, the mouse was placed on a heating pad and monitored until it had fully recovered from anesthesia.
7
Assessment of Pediatric Amblyopia
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A standard logarithmic visual acuity chart was used to check corrected visual acuity in the naked eyes of those who could cooperate with the examination. Routine anterior segment examination and fundoscopic examination were performed on all patients to exclude other organic ocular lesions. Cycloplegic refraction was used to assess refractive errors. Cycloplegia was achieved using four drops of compound tropicamide eye drops (mixture of 0.5% tropicamide and 0.5% phenylephrine eye drops; Santen Pharmaceutical Co., Ltd., Osaka, Japan), administered approximately 5 min apart. Cycloplegic autorefraction was measured at 30 min after the last drop using a desktop autorefractor (KR-8800; Topcon Corporation, Tokyo, Japan). Three readings with 0.25 Diopter (D) or less apart in both the spherical and cylindrical components were averaged. Then, cycloplegic retinoscopy was performed by an experienced optometrist. According to the consensus of experts on the Prevention and Treatment of Amblyopia in Chinese Children (2021) [7 (link)], to exclude ocular organic disease, the normal lower limit of vision is 0.5 for children aged 3–5 years and 0.7 for children aged ≥6 years. If the corrected visual acuity is lower than this limit or if the difference of visual acuity is two rows or more, amblyopia is considered.
8
Laser-Induced Choroidal Neovascularization Model
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The laser photoagglutination-induced CNV model was performed in accordance with previous reports [21 (link)]. Sodium pentobarbital (1%, Sigma, St. Louis, MO, USA; P3761) was first administered intraperitoneally to male C57BL/6J mice to anesthetize them; compound tropicamide eyedrops (Santen Pharmaceutical, Osaka, Japan) and oxybuprocaine hydrochloride eyedrops (0.4%, Santen Pharmaceutical) were then applied to the eyes. An incident 532 nm laser (Twin, Quantel, France) was shone into the right eye of each animal while their pupils were dilated, inducing burns 1.5–2 disc diameters away from the optic nerve (75 μm, 100 ms, and 120 mV). As long as a vaporization bubble did not form with hemorrhage, laser spots that ruptured Bruch’s membrane were considered valid, and the CNV lesions were employed for analysis. Immediately after laser photocoagulation, animals were randomly divided into four groups.
9
Pupil Dilation and ICL Implantation
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Before surgery, the eyes were instilled with compound tropicamide eye drops (Santen, Japan) 4 times to expand the pupil diameter to greater than 7 mm. Oxybuprocaine hydrochloride (Santen, Japan) eye drops were applied 4 times as topical anaesthesia. A small OVD amount (1.7% sodium hyaluronate, Shandong Bausch and Lomb Freda Company) and balanced salt solution (BSS, Alcon, USA) were injected into the ICL implant container. The injector was properly installed and placed in BSS for later use. An assisted right-side incision was made, through which an appropriate OVD amount was injected into the AC. A 3.0 mm clear corneal main incision was made, typically placed on the steep axis of the corneal curvature, to release corneal astigmatism. The ICL was inserted from the main incision of the AC, and a sufficient OVD amount was injected into the front surface of the ICL to maintain the operating space. After gently tucking the footplate beneath the iris and adjusting the ICL to a central position, the OVD was manually irrigated from the AC using BSS by an injection needle attached to a syringe. Finally, the incisions were made watertight.
10
Retinal Function Assessment in Mice
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After 7 days of MNU administration, the mice were examined by ERG. After 24 hr of dark adaptation, mice were anesthetized by ip injection of chlorpromazine (15 mg/kg) and ketamine 60 mg/kg (Pfizer, USA). Compound tropicamide eye drops (Santen Pharmaceuticals, Japan) were administered to dilate the pupils, and the corneas were anesthetized with cocaine hydrochloride eye drops (Santen Pharmaceutical Co., Japan). On the recording stage under dim red light, the recording electrode was placed on the limbus, and the reference electrode and the ground electrode were placed on the cheek and tail, respectively. The b-wave amplitude and latency were recorded with a single flash stimulation.
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