To evaluate retinal morphology and BRB integrity in vivo, OCT, and FA were performed simultaneously in living mice following our previous publication (25 (link), 52 ). Mice were put to sleep using 2% isoflurane anesthetic, followed by dilation of their pupils using 1% tropicamide eye drop and Genteal gel was applied liberally to keep the eye moist during imaging. Each mouse was evaluated by OCT and FA after being placed on the platform of the Phoenix Micron III retinal imaging microscope supplemented with an OCT imaging device (Phoenix Research Laboratories, Pleasanton, CA). Mice were injected intraperitoneal with fluorescein sodium 10 to 20μL 10% (Apollo Ophthalmics, Newport Beach, CA), followed by rapid acquisition of fluorescent images ensued for ~5 minutes. Leakage of fluorescein manifests as unclear vascular borders increasing progressively to diffusing hazy fluorescence.
Phoenix micron 3
Manufactured by Phoenix Pharmaceuticals
Sourced in United States
The Phoenix Micron III is a high-precision laboratory instrument designed for accurate measurements and analysis. It features advanced technology to ensure reliable and consistent results. The core function of the Phoenix Micron III is to provide precise data for scientific research and experiments.
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7 protocols using phoenix micron 3
1
In Vivo Retinal Morphology Evaluation
2
Retinal Vascular Permeability Assessment in Rats
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Retinal vascular permeability in living animals was assessed as described before (34 (link)). Briefly, rats were anesthetized (ketamine, 100 mg/kg; xylazine, 30 mg/kg; and acepromazine, 10 mg/kg). Pupils were dilated using 1% tropicamide (Bausch & Lomb, Rochester, NY), and Goniovisc 2.5% (hypromellose; Sigma Pharmaceuticals, LLC, Monticello, IA) was applied liberally to retain surface moisture during imaging. Each animal was placed on the imaging platform of the Phoenix Micron III retinal imaging microscope (Phoenix Research Laboratories, Pleasanton, CA), and an intraperitoneal injection (80–100 μL) of fluorescein sodium (10% Lite) (Apollo Ophthalmics, Newport Beach, CA) was administered. Rapid acquisition of fluorescent images ensued for ∼5 min. For a quantitative evaluation of blood-retinal barrier (BRB) integrity, we assessed albumin extravasation in rats after perfusion (34 (link),35 (link)). Serum albumin levels were measured in the perfused retinal tissue by Western blot using anti-rat albumin antibody (Cell Signaling Technology, Danvers, MA).
3
Retinal Vasculature and Permeability Evaluation
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To evaluate the retinal vasculature and permeability in vivo, one and six weeks after oral gavage/infection with Pg and biofilm, mice were anesthetized using intramuscular injection of the rodent anesthesia cocktail described above [74 (link)]. The pupils were dilated with 1% tropicamide eye drops, and the mice were placed on the imaging platform of the Phoenix Micron III retinal-imaging microscope (Phoenix Research Laboratories, Pleasanton, CA, USA). Goniovisc 2.5% (hypromellose; Sigma Pharmaceuticals, LLC, Monticello, IA, USA) was applied generously to maintain moisture in the eyes during imaging. Mice were administered 10 to 20 μL 10% fluorescein sodium (Apollo Ophthalmics, Newport Beach, CA, USA), and rapid acquisition of fluorescent images ensued for ∼5 min. Fluorescein leakage manifests as indistinct vascular borders progressing to diffusely hazy fluorescence. Fluorescein leakage (CNV area) was compared between different groups by quantifying the fluorescence intensities collected after 1, 2, and 3 min following fluorescein injection using ImageJ software (National Institutes of Health, Bethesda, MD, USA).
4
In Vivo Retinal Vascular Imaging
5
Mouse Fundus and OCT Imaging
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Fundus imaging and SD-OCT were done as previously described [58 (link),59 (link)]. Briefly, 2% isoflurane was used to anesthetize the mice, and their pupils were dilated using 1% tropicamide eye drops. Mice were placed on the imaging platform of the Phoenix Micron III retinal imaging microscope, supplemented with an OCT imaging device (Phoenix Research Laboratories, Pleasanton, CA). Genteal gel was applied during imaging to keep the eye moist. Four images were taken for each mouse eye.
6
Characterization of Healthy Ocular Tissues
7
Multimodal Imaging of Retinal RPE
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To evaluate retinal RPE structure and function in vivo, OCT and FA were performed simultaneously as described previously with some modifications [41 (link)]. Briefly, mice were anesthetized using 2% isoflurane and their pupils were dilated using 1% tropicamide eye drop. Each mouse was then placed on the imaging platform of the Phoenix Micron III retinal imaging microscope supplemented with OCT imaging device (Phoenix Research Laboratories, Pleasanton, CA). Genteal gel was applied liberally to keep the eye moist during imaging. Mice were administered 10 to 20 μL 10% fluorescein sodium (Apollo Ophthalmics, Newport Beach, CA), and rapid acquisition of fluorescent images ensued for ∼5 minutes. Fluorescein leakage manifests as indistinct vascular borders progressing to diffusely hazy fluorescence.
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