Larvae were immobilized dorsal up in 3% methylcellulose and imaged with Optical Coherence Tomography (OCT) (~840nm; Leica Bioptigen R2210 Spectral Domain Ophthalmic Imaging System). After imaging, larvae were rinsed and transferred into petri dishes to recover. OCT scans were analyzed using Bioptigen software (InVivoVue; Bioptigen, Research Triangle Park, NC), FIJI, and MetaMorph.
Invivovue
Manufactured by Bioptigen
Sourced in Germany
InVivoVue is a high-performance optical coherence tomography (OCT) system designed for preclinical research applications. It provides real-time, high-resolution, three-dimensional imaging of biological samples, including small animal models.
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Lab products found in correlation
7 protocols using invivovue
1
Larval OCT Imaging Protocol
2
Optical Coherence Tomography of Myo7a Mice
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Prior to Optical Coherence Tomography (OCT) analysis, pupils of mice were dilated (atropine 1%, phenylephrine 2.5%). Mice were anesthetized, and retinal cross-section images were obtain with a spectral domain (SD) OCT system (Bioptigen, Inc., Durham, NC, United States). Post-imaging ONL measurements were performed with commercial software (InVivoVue, Bioptigen, Inc.), as previously described (Pang et al., 2011 (link)). Equal numbers of males and females were used for each genotype (n = 10–11/genotype). Two-way ANOVA with Bonferroni’s multiple comparisons tests were used to analyze OCT measurements in Myo7a+/+, Myo7a+/–, and Myo7a–/– mice. Significance was defined as a P-value <0.05.
3
Intraoperative OCT-Guided Subretinal Therapy
4
In vivo Retinal Imaging and Layer Analysis
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In vivo retinal images were acquired using an Envisu UHR2200 (Bioptigen, Durham, NC) with theoretical axial resolution in the tissue of 1.75 µm. Mice were anesthetized, eyedrops were used to dilate the pupils and keep the eyes hydrated as described above. Mice were positioned to have the eye look directly into the lens, and then to align the optic nerve head to be on the center of the OCT scan horizontally and vertically. A rectangular scan with a 1.8 mm length and width, 0° angle, 0 mm horizontal and vertical offsets 1000 lines of A-scans/B-scans, 100 B-scans, 10 frames/B-scan, 80 lines of inactive A-scans/B-scan, and 1 volume was captured. Ten-frame OCT images were averaged using Bioptigen InVivoVue® and the averaged image was subjected to auto-segmentation for retinal layer thickness measurement using Bioptigen Diver® V. 3.4.4 software.
5
Retinal Layer Thickness Measurement
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The individual retinal layers were identified using human retina OCT images and domestic pig OCT data published by the Gloesmann et al.14 (link) and Staurenghi et al.15 Nine retinal layers were measured, including the nerve fiber layer, ganglion cell layer, inner plexiform layer, inner nuclear layer, outer plexiform layer, outer nuclear layer, photoreceptor inner segment, photoreceptor outer segment, and retinal pigment epithelium (RPE). On the OCT scans, manual retinal thickness measurements were performed in the middle of the horizontal line scans for each individual retinal layer by two masked, independent graders using the caliper tool provided by the software (Bioptigen InVivoVue, version 2.4.34; Fig. 3 ). After determining the corresponding histologic section, the area and length of each retinal layer were measured using ImageJ (version 1.48, National Institutes of Health, Bethesda, MD). The average thickness then was calculated using these measurements (e.g., area divided by length).
6
In Vivo Retinal Imaging of Mouse Models
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In vivo retinal imaging was performed as previously described in detail by DeRamus et al. [15 (link)], using a Bioptigen Model 840 Envisu Class-R high-resolution SD-OCT instrument (Bioptigen/Leica, Inc.; Durham, NC, USA). Data were collected from DhddsK42E/K42E and WT mice at postnatal day (PN) 1 (KI, n = 5; WT, n = 9), 2 (KI, n = 4; WT, n = 8), 3 (KI, n = 5; WT n = 3), 8 (KI, n = 4; WT n = 5), and 12 months (mos) (KI, n = 3; WT n = 3) to assess retinal structure. Layer thicknesses were determined manually using Bioptigen InVivoVue® and Bioptigen Diver® V. 3.4.4 software and the data were analyzed and graphed using Microsoft Excel software.
7
In Vivo Retinal Imaging in Mice
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Pupils were dilated and mice were anesthetized using ketamine-xylazine (50–5 mg/kg) as previously described [28 (link)]. In vivo imaging was performed on a Bioptigen Model 840 Envisu Class-R high-resolution SD-OCT instrument (Bioptigen/Leica, Inc; Durham, NC, USA). Retinal layer thicknesses were determined manually using Bioptigen InVivoVue® and Bioptigen Diver® V. 3.4.4 software. Data were analyzed and presented using Microsoft Excel software.
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