Fundus fluorescein angiography (FFA) in mice was performed according to a previous study (24 (link)). Mice were anesthetized by ketamine/xylazine intraperitoneal injection and pupils were dilated with topical 1% tropicamide (Santen, Osaka, Japan). Before the procedure, 100 μL of 10% fluorescein sodium (Alcon, US) was injected intraperitoneally. We then placed the mice at a 170° angle with the head closer to the operator. Fluorescent fundus images were acquired within 5 minutes after fluorescein sodium injection. The vascular leakage area of the CNV lesions was determined by measuring the hyperfluorescent area with ImageJ Software.
Fluorescein sodium
Manufactured by Alcon
Sourced in United States, Switzerland, Germany
Fluorescein sodium is a water-soluble dye used in various laboratory applications. It is a bright yellow-green fluorescent compound that can be detected under ultraviolet or blue-light illumination. The compound is commonly used as a tracer or staining agent in various diagnostic and research procedures.
Automatically generated - may contain errors
Lab products found in correlation
Fluorescein, Alcon (3 mentions)
Micron IV, Phoenix Pharmaceuticals (2 mentions)
Micron III retinal imaging system, Phoenix Pharmaceuticals (2 mentions)
Tropicamide, Santen (1 mentions)
Tropicamide solution, Alcon (1 mentions)
Gonak, Akorn (1 mentions)
SD-OCT, Heidelberg Engineering (1 mentions)
Model TRC 50 IA, Topcon (1 mentions)
TRC-50EX, Topcon (1 mentions)
Plan Apo λ, Nikon (1 mentions)
TUNEL kit, Roche (1 mentions)
TRC-501X, Topcon (1 mentions)
Heidelberg Spectralis device, Heidelberg Engineering (1 mentions)
Melatonin, Merck Group (1 mentions)
Colivelin, R&D Systems (1 mentions)
Heidelberg Retina Angiograph 2, Heidelberg Engineering (1 mentions)
Spectralis HRA+OCT, Heidelberg Engineering (1 mentions)
Micron IV system, Phoenix Pharmaceuticals (1 mentions)
Micron III, Phoenix Pharmaceuticals (1 mentions)
Tropicamide, Sandoz (1 mentions)
40 protocols using Fluorescein sodium
1
Fundus Fluorescein Angiography in Mice
2
Evaluating Laser-Induced Choroidal Neovascularization
Fluorescein angiography (FA) was performed on anesthetized animals, using an Optos 200Tx ultra-wide field retinal imaging system (Optos PLC, Dunfermline, Scotland, UK), 14 days following laser photocoagulation. Photographic images were captured after 0.4 mL 10% fluorescein sodium (Alcon Laboratories, Inc., Fort Worth, TX) was injected into the peritoneum. Two masked retinal specialists, not involved in laser photocoagulation or angiography, evaluated FA images. Lesions were graded using a previously established grading system [24] (link). Grade 0 was given if no hyperfluorescence was present in the early and late phases. Grade 1 was given if hyperfluorescence without leakage was present in the early or late phase. Grade 2A was given if hyperfluorescence in the early or middle phase and leakage in the late phase was observed. With this grade, late leakage did not extend beyond the treated areas. Grade 2B was given if bright hyperfluorescence in the middle phase and leakage beyond treated areas in the late phase existed. Grade 2B lesions were regarded as clinically significant [24] (link). Grade 0 lesions were excluded from analyses because the laser injury may not have been sufficient to induce CNV.
3
Analyzing Retinal Vasculature in P17 Mice
P17 mice were lightly anesthetized by IP injection of 2.5% tribromoethanol (0.020 mL/g; Sigma-Aldrich). Before the procedure, the eyes were dilated with one drop of tropicamide solution (Alcon, Fort Worth, TX, USA) and lubricated with hypromellose ophthalmic demulcent solution (Gonak; Akorn Pharmaceuticals, Lake Forest, IL, USA). We then placed the mice on a custom heated stage for imaging. Fundus fluorescein angiography (FFA) was performed immediately after fluorescein sodium (13 mL/kg in saline; Alcon) injection via the tail vein. Images were captured every 30 seconds for 5 minutes using a retinal imaging system (OPTO-RIS; Optoprobe Science, Burnaby, BC, Canada). Fluorescence intensity was calculated by multiplying the mean retinal vessel intensity by vessel area.15 (link),22 (link) Retinal vein width for each mouse was measured by selecting two points horizontally at the edge of the vein.23 (link) Retinal arterial tortuosity was calculated as the ratio of actual vessel length to the straight linear length from the optic disc to the edge of FFA images.23 (link),24 (link) Average retinal vein width and artery tortuosity were used to represent each mouse. All parameters were measured using ImageJ software.25 (link) All of the captures were normalized to the WT in RA group.
4
Retinal Artery Imaging in Mice
We used an intraperitoneal injection of 2.5% tribromoethanol (0.020 mL/g; Sigma-Aldrich, Dorset, UK) to anesthetize the mice. One compound-tropicamide eye drop was used to dilate each pupil, and then the eye was treated with ophthalmic gel (hypromellose 2.5% ophthalmic-demulcent solution; Gonak; Akorn, Lake Forest, IL, USA). The mice were subsequently given a tail vein injection of fluorescein sodium (13 mL/kg in saline; Alcon, TX, USA). After that, for five minutes, we used a retinal imaging equipment (OPTO-RIS; Optoprobe Science, Burnaby, BC, Canada) to take pictures of the retinal arteries every thirty seconds. The branch architecture and pulsatile activity of arteries were used to identify them. In order to determine the arteriovenous ratio for each mouse, we selected an identifiable anatomical site that was two optic-disc diameters from the optic disc. ImageJ (Rasband; NIH) software was used to compare measurements [25 (link)].
5
Fluorescein Angiography and OCT for Optic Nerve Edema
Fundus fluorescein angiography (FFA), and spectral domain optical coherence tomography (SD-OCT) were conducted on days 0, 1, 3 and 7 following treatment to observe the progress of the optic nerve head edema and the related retinal response. A solution of 10% Fluorescein sodium (Alcon Laboratories Inc. Switzerland) was injected intraperitoneally. The angiographs were recorded following the change in coloration of the conjunctiva that appeared yellow in color (Heidelberg Engineering, Heidelberg,Germany). Retinal structure and retinal thickness were measured using SD-OCT (Heidelberg Engineering). Retinal scans were centered on the optic disc in both control and injured eyes.
6
Fundus Imaging and Fluorescein Angiography
Color fundus photography and FFA were performed on a fundus imaging system (MicronIV; PHOENIX, USA) at 4 weeks after vitreous injection. After animals were anesthetized and pupils were dilated, color fundus photography was performed. FFA images were recorded 5 min after intraperitoneal injection of fluorescein sodium (10%, 0.3 ml; Alcon).
7
Fundus Fluorescein Angiography in Mice
After anesthesia with the intraperitoneal application of a mixture containing ketamine (80 mg/kg) and xylazine (10 mg/kg), the pupils were dilated with 1% tropicamide and the cornea was kept moist using 1% carboxymethylcellulose sodium eye drops. The mice were injected intraperitoneally with 10% fluorescein sodium (Alcon, USA) at a dose of 1.5 ml/kg. The assay of fundus fluorescein angiography (FFA) was performed using a Micro-III retinal image system (Phoenix Research Labs Inc., USA). The acquisition of fundus images was performed 5 mins after the injection of fluorescein sodium at a rate of 4FPS.
8
Fluorescein Angiography for SLRNPA vs. DR
To distinguish SLRNPA from DR, FFA was performed using the Spectralis Heidelberg Engineering system. After standard intravenous injection of 5 mL 10% (w/v) fluorescein sodium (Alcon Laboratories Inc.), FFA images were obtained over a 55° area 45 s and 2 and 10 min later (the early, middle, and late phases of angiography, respectively). Central images were centered on the macula; we also obtained superior, temporal, nasal, and inferior images to allow clear visualization of the peripheral edges of the retinal vasculature.
9
Fundus Fluorescein Angiography Grading Protocol
Fundus fluorescein angiography was performed 7 days postlaser by intraperitoneal injection of 50 μL of 25% fluorescein sodium (Alcon, Fort Worth, TX, USA). Fundus images were taken using a digital fundus camera (Model TRC 50 IA; Topcon, Paramus, Japan). FFA was qualitatively and quantitatively evaluated by two blind groups of observers. The laser-induced lesions were graded based on the observed fluorescein leakage and then divided into the following four categories as described by Semkova [20 (link)]. Volumes of grade 4 laser spots were evaluated by image J software.
10
Retinal Vessel Diameter Estimation
On the second day after delivery, the rats were anesthetized with sodium pentobarbital (50 mg/kg intraperitoneally), the pupils were dilated with 1% tropicamide, and the cornea was kept moist using 1% carboxymethylcellulose sodium eye drops. The rats were injected intraperitoneally with 10% fluorescein sodium (Alcon, USA) at a dose of 1.5 ml/kg. Images were recorded 3 to 240 seconds after injection. Fundus fluorescein angiography (FFA) was performed with a digital fundus camera (TRC-50EX; Topcon, Tokyo, Japan). Final fundus photographs were used for estimating the arteriolar and venular diameter. The diameter of retinal vessels was estimated by the methodology of Gupta et al.34 (link). Diameter measurements were obtained at three different locations along each vessel near optic disk, with an average of three measurements.
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