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* These authors contributed equally
The protocol presented here details the procedures of data collection and data analysis for image-guided optical coherence tomography (OCT) and demonstrates its application in multiple rodent models of ocular diseases.
Ocular diseases, such as age-related macular degeneration, glaucoma, retinitis pigmentosa, and uveitis, are always accompanied by retinal structural changes. These diseases affecting the fundus always exhibit typical abnormalities in certain cell types in the retina, including photoreceptor cells, retinal ganglion cells, cells in the retinal blood vessels, and cells in the choroidal vascular cells. Noninvasive, highly efficient, and adaptable imaging techniques are required for both clinical practice and basic research. Image-guided optical coherence tomography (OCT) satisfies these requirements because it combines fundus photography and high-resolution OCT, providing an accurate diagnosis of tiny lesions as well as important changes in the retinal architecture. This study details the procedures of data collection and data analysis for image-guided OCT and demonstrates its application in rodent models of choroidal neovascularization (CNV), optic nerve crush (ONC), light-induced retinal degeneration, and experimental autoimmune uveitis (EAU). This technique helps researchers in the eye field to identify rodent retinal structural changes conveniently, reliably, and tractably.
Ocular diseases affecting the fundus always exhibit typical abnormalities in certain cell types in the retina, such as photoreceptor cells, retinal ganglion cells, cells in the retinal blood vessels, and cells in the choroidal blood vessels, which may subsequently influence the visual acuity of patients1. To avoid irreversible visual impairment, timely diagnoses and appropriate treatments are required1. Optical coherence tomography (OCT) has been widely used in the clinic to evaluate a range of ocular diseases, including age-related macular degeneration, retinitis pigmentosa, glaucoma, uveitis, and retinal detachment, among others2,3,4. This kind of noninvasive, highly efficient, and adaptable imaging technique is also needed for the timely evaluation of the disease conditions in experimental animals5,6,7,8,9,10.
Image-guided optical coherence tomography (OCT) uses interferometry to produce cross-sectional images of animal retinas at 1.8 µm longitudinal resolution and 2 µm axial resolution. It has at least three advantages in the investigation of retinal architectural changes2,3,4,5,6,7,8,9,10. First, it is a noninvasive technique that allows researchers to dynamically follow the location of interest in the same animal retina5,6,7,8,9,10. Second, this trait substantially reduces the sample size for every experiment3. Meanwhile, it saves considerable time and effort in research projects2,3,4,5,6,7,8,9,10. Third, image-guided OCT acquires colorful fundus images while capturing OCT images, thus providing accurate and reliable results for users.
This manuscript describes the procedures of image collection and data analysis for image-guided OCT and elaborates on its application in mouse and rat models of choroidal neovascularization (CNV)11,12, optic nerve crush (ONC)13,14,15,16, light-induced retinal degeneration17,18,19,20,21, and experimental autoimmune uveitis (EAU)22,23. With this versatile technique, researchers can capture high-resolution OCT images as well as fundus images conveniently and efficiently.
All the animal procedures conformed to the Association for Research on Vision and Ophthalmology's statement on the Use of Animals in Ophthalmic and Vision Research and were approved by the Institutional Animal Care and Use Committee of Wenzhou Medical University (WMU). The rats and mice were given free access to water and food with an environmental light intensity of 18 lux on a 12 h dark/light cycle.
1. Preparation of the ocular animal models
2. OCT module setup
3. Animal preparation for OCT experiments
4. Image-guided OCT Imaging
NOTE: The software interface was divided into three parts: brightfield image, OCT control tabs, and OCT display (Figure 2).
5. Thickness measurement and quantitative analysis
NOTE: This OCT has built-in analysis software. OCT images can be segmented and analyzed using this software (Figure 3).
Image-guided OCT can be used to monitor the development of the laser spot in laser-induced choroidal neovascularization (CNV) in mice. As shown in Figure 1, the newborn blood vessels passed through Bruch's membrane as well as the retinal pigment epithelium (RPE) layer and formed a fibrotic scar after laser injury11,12. This lesion spot could be captured under either full-size scanning (Figure 4A) or ...
This protocol provides instructions for the image collection and thickness measurement of image-guided OCT. By demonstrating the four most popular rodent models of ocular diseases, the researchers found that image-guided OCT provided excellent performance in examining drastic retinal structural alterations. In fact, with high-resolution images, tiny lesions can be found easily in OCT images as well. With the aid of image-guided OCT, a group in the laboratory also found abnormal hyperreflectivity spots within the OPL in a...
None of the authors have any conflicts of interest to disclose.
The authors thank the members of the State Key Laboratory of Ophthalmology, Optometry, and Vision Science for their technical support and useful comments regarding the manuscript. This work was supported by grants from the National Natural Science Foundation of China (82101169, 81800857, 81870690), the Zhejiang Provincial Natural Science Foundation of China (LGD22H120001, LTGD23H120001, LTGC23H120001), the Program of Wenzhou Science and Technology Bureau of China (Y20211159), the Guizhou Science and Technology Support Project (Qiankehezhicheng [2020] 4Y146) and the Project of State Key Laboratory of Ophthalmology, Optometry and Vision Science (No. K03-20220205).
Name | Company | Catalog Number | Comments |
BALB/c mouse | Beijing Vital River Laboratory Animal Technology Co., Ltd | Animal model preparations | |
C57BL/6JNifdc mouse | Beijing Vital River Laboratory Animal Technology Co., Ltd | Animal model preparations | |
Carbomer Eye Gel | Fabrik GmbH Subsidiary of Bausch & Lomb | Moisten the cornea | |
Complete Freund’s adjuvant | Sigma | F5881 | EAU experiment |
Experimental platform | Phoenix Technology Group | Animal model preparations | |
hIRBP161-180 | Shanghai Sangon Biological Engineering Technology & Services Co., Ltd. | EAU experiment | |
Ketamine | Ceva Sante Animale | General anesthesia | |
Laser box | Haag-Streit Group | Merilas 532α | Animal model preparations |
Lewis rat | Beijing Vital River Laboratory Animal Technology Co., Ltd | Animal model preparations | |
Mycobacterium Tuberculosis H37RA | Sigma | 344289 | EAU experiment |
Phoneix Micron IV with image-guided OCT and image-guided laser | Phoenix Technology Group | Animal model preparations | |
Tissue forceps | Suzhou Mingren Medical Instrument Co., Ltd | MR-F101A-5 | Animal model preparations |
Tropicamide Phenylephrine Eye Drops | SANTEN OY, Japan | Eye dilatation | |
Vannas scissors | Suzhou Mingren Medical Instrument Co., Ltd | MR-S121A | Animal model preparations |
Xylazine | Ceva Sante Animale | General anesthesia |
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