Name: in vivo structural assessments of ocular disease in rodent models using optical coherence tomography
Uploaded: 2020-07-24T14:00:00
Description: Watch this Scientific Journal Video about In vivo Structural Assessments of Ocular Disease in Rodent Models using Optical Coherence Tomography at JoVE.com

This work was supported by the Department of Veterans Affairs Rehab R&#38;D Service Career Development Awards (CDA-1, RX002111; CDA-2; RX002928) to RSA, Merit Award (RX002615) and Research Career Scientist Award (RX003134) to MTP, Career Development Award (CDA-2, RX002342) to AJF, EY028859 to MTP, NEI Core Grant P30EY006360, Research to Prevent Blindness, and Foundation Fighting Blindness.

All procedures described were approved by the Atlanta Veterans Affairs Institutional Animal Care and Use Committee and conformed to the National Institutes of Health guide for the care and use of laboratory animals (NIH Publications, 8th edition, updated 2011).
NOTE: The SD-OCT system used to develop the protocol below is described in the Table of Materials. While some of the procedures are specific to this particular system, the overall approach can be adapted for ...

<ol>
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A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Altered+ocular+parameters+from+circadian+clock+gene+disruptions.">Altered ocular parameters from circadian clock gene disruptions.</a> PLoS One. 14 (6), 0217111 (2019).</li><li>Chakraborty, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Circadian+rhythms,+refractive+development,+and+myopia.">Circadian rhythms, refractive development, and myopia.</a> Ophthalmic &amp; Physiological Optics. 38 (3), 217-245 (2018).</li><li>Davies, E. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retinal+ganglion+cell+layer+volumetric+assessment+by+spectral-domain+optical+coherence+tomography+in+multiple+sclerosis:+application+of+a+high-precision+manual+estimation+technique.">Retinal ganglion cell layer volumetric assessment by spectral-domain optical coherence tomography in multiple sclerosis: application of a high-precision manual estimation technique.</a> Journal of Neuro-ophthalmology. 31 (3), 260-264 (2011).</li><li>Carnevali, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+coherence+tomography+angiography+analysis+of+retinal+vascular+plexuses+and+choriocapillaris+in+patients+with+type+1+diabetes+without+diabetic+retinopathy.">Optical coherence tomography angiography analysis of retinal vascular plexuses and choriocapillaris in patients with type 1 diabetes without diabetic retinopathy.</a> Acta Diabetologica. 54 (7), 695-702 (2017).</li><li>Springelkamp, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Population-based+evaluation+of+retinal+nerve+fiber+layer,+retinal+ganglion+cell+layer,+and+inner+plexiform+layer+as+a+diagnostic+tool+for+glaucoma.">Population-based evaluation of retinal nerve fiber layer, retinal ganglion cell layer, and inner plexiform layer as a diagnostic tool for glaucoma.</a> Investigative Ophthalmology &amp; Visual Science. 55 (12), 8428-8438 (2014).</li><li>Allen, R. 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High resolution imaging of ocular structures in vivo allows for the assessment of retinal and ocular changes over time. In this protocol, SD-OCT was demonstrated to capture differences in ocular structures in vivo in models of retinal degeneration, glaucoma, diabetic retinopathy, and myopia.
The most critical aspect when performing SD-OCT is obtaining a clear image of the retina or other ocular structure of interest. It is important to take time to make sure the retina is perfectly centered an...

Spectral-domain optical coherence tomography (SD-OCT) is a precise, high-resolution imaging modality that allows clinicians and researchers to examine ocular structures noninvasively. This imaging technique is based on interferometry to capture three-dimensional retinal images in vivo on a micrometer scale1,2. It has become one of the most frequently used imaging modalities in vision research and in the clinic due to the easy detection and accuracy of pathological features such as structural defects and/or thinning of retinal layers and subretinal fluid3. In research using animal models of vision-related disorders, SD-OCT has provided essential noninvasive analyses of relationships between structure and function and their histopathological origins4. Due to its resolution (up to 2-3 microns, depending on the depth into the eye5), SD-OCT has the capability to detect even small changes in retinal layer thickness. This type of analysis can provide essential information for disease progression and assess the efficacy of neuroprotective methods and treatments for vision-related disorders.
SD-OCT is a noninvasive alternative to examining structure histologically, and the two have been shown to be correlated6. While SD-OCT does not reach cellular resolution, it does allow for longitudinal studies in animals. This is advantageous because disease progression can be tracked in individual animals over time as opposed to having to euthanize animals at specific time points. As imaging techniques continue to improve, SD-OCT technology will also progress, providing enhanced image quality as well as the capability to assess biological processes such as retinal blood vessel function in fine detail. Even since its advent in 1991, SD-OCT technology has seen huge advances in resolution, speed, and sensitivity7.
The present study utilizes an SD-OCT system to quantify changes in retinal layers in rodent models of retinal degeneration, glaucoma, and diabetic retinopathy. The SD-OCT system used here is a Fourier-domain OCT-system that utilizes low-power, near-infrared light to acquire, process, and store depth-resolved images in real time. The SD-OCT system has extended depth-imaging capability in the 800 nm wavelength band, providing 8 mm depth and 4 &#181;m resolution. In Fourier domain detection, the interference signal between scattered light from the tissue and a reference path is Fourier transformed to construct axial scans and/or axial depth profiles of scattered intensity8. For the studies here, the OCT beam is scanned over the desired retinal structure while serially acquiring axial scans. Typically, a scan pattern acquires the two-dimensional grid (B-Scans) as a collection of linear one-dimensional scan lines (A-Scans), which correspond to 2D cross-sectional images using a raster scan pattern. For studies focused on myopia in mice, this system is also used to measure dimensions of ocular structures (e.g., cornea thickness, lens thickness, vitreous chamber depth, and axial length).
The current system allows users to design their own protocols, creating scans that can be tailored and selected based on the ocular structures of interest. The principal scans featured in these user defined protocols makes this imaging technique user-friendly. For image analyses, we have developed customized programming in a mathematic modeling program. SD-OCT is a powerful tool to non-invasively identify and quantify pathomorphological changes in ocular structures and monitor vision-related disease progression.

<table>
	<tbody>
		<tr>
			<td>1% tropicamide</td>
			<td>Sandoz</td>
			<td>Sandoz #6131403550; NDC- 24208-585-59</td>
			<td></td>
		</tr>
		<tr>
			<td>0.5% tetracaine</td>
			<td>Alcon</td>
			<td>NDC 0065-0741-12</td>
			<td></td>
		</tr>
		<tr>
			<td>AIM-RAS G3 120 V</td>
			<td>Leica Bioptigen</td>
			<td>90-AIMRAS-G3-120</td>
			<td>Specialized platform to hold the OCT Scanner Head for mice</td>
		</tr>
		<tr>
			<td>Celluvisc gel</td>
			<td>REFRESH CELLUVISC</td>
			<td>#4554; NDC-0023-4554-30</td>
			<td></td>
		</tr>
		<tr>
			<td>G3 18 mm Telecentric Lens</td>
			<td>Leica Bioptigen</td>
			<td>90-BORE-G3-18</td>
			<td></td>
		</tr>
		<tr>
			<td>G3 Mouse Lens</td>
			<td>Leica Bioptigen</td>
			<td>90-BORE-G3-M</td>
			<td></td>
		</tr>
		<tr>
			<td>G3 Rat Lens</td>
			<td>Leica Bioptigen</td>
			<td>90-BORE-G3-R</td>
			<td></td>
		</tr>
		<tr>
			<td>heating pad</td>
			<td>Fabrication</td>
			<td>11-1130</td>
			<td></td>
		</tr>
		<tr>
			<td>InVivoVue software</td>
			<td>Leica Bioptigen</td>
			<td></td>
			<td>Specialized software that pairs with the Leica Bioptigen SD-OCT system</td>
		</tr>
		<tr>
			<td>MATLAB</td>
			<td>Mathworks</td>
			<td></td>
			<td>mathematical modeling program</td>
		</tr>
		<tr>
			<td>Mouse/Rat Kit</td>
			<td>Leica Bioptigen</td>
			<td>90-KIT-M/R</td>
			<td>Mouse/rat rodent alignment system</td>
		</tr>
		<tr>
			<td>saline</td>
			<td>ADDIPAK</td>
			<td>200-39</td>
			<td></td>
		</tr>
		<tr>
			<td>System Envisu R4300 VHR 120 V</td>
			<td>Leica Bioptigen</td>
			<td>90-R4300-V1-120</td>
			<td>SD-OCT system</td>
		</tr>
	</tbody>
</table>

in vivo structural assessments of ocular disease in rodent models using optical coherence tomography

Spectral-domain optical coherence tomography (SD-OCT) is useful for visualizing retinal and ocular structures in vivo. In research, SD-OCT is a valuable tool to evaluate and characterize changes in a variety of retinal and ocular disease and injury models. In light induced retinal degeneration models, SD-OCT can be used to track thinning of the photoreceptor layer over time. In glaucoma models, SD-OCT can be used to monitor decreased retinal nerve fiber layer and total retinal thickness and to observe optic nerve cupping after inducing ocular hypertension. In diabetic rodents, SD-OCT has helped researchers observe decreased total retinal thickness as well as decreased thickness of specific retinal layers, particularly the retinal nerve fiber layer with disease progression. In mouse models of myopia, SD-OCT can be used to evaluate axial parameters, such as axial length changes. Advantages of SD-OCT include in vivo imaging of ocular structures, the ability to quantitatively track changes in ocular dimensions over time, and its rapid scanning speed and high resolution. Here, we detail the methods of SD-OCT and show examples of its use in our laboratory in models of retinal degeneration, glaucoma, diabetic retinopathy, and myopia. Methods include anesthesia, SD-OCT imaging, and processing of the images for thickness measurements.

SD-OCT is considered successful if high quality images are obtained such that ocular dimensions can be reliably measured. Here, a variety of uses of SD-OCT are illustrated using models of retinal degeneration, glaucoma, diabetic retinopathy, and myopia.
In a light-induced retinal degeneration (LIRD) model, exposure to bright light (10,000 lux) induces degeneration of photoreceptor cells in the retina9. Representative SD-OCT images reveal a thinner outer nuclear layer, w...

Watch this Scientific Journal Video about In vivo Structural Assessments of Ocular Disease in Rodent Models using Optical Coherence Tomography at JoVE.com

In vivo Structural Assessments of Ocular Disease in Rodent Models using Optical Coherence Tomography

Here, we describe the use of spectral-domain optical coherence tomography (SD-OCT) to visualize retinal and ocular structures in vivo in models of retinal degeneration, glaucoma, diabetic retinopathy, and myopia.

Spectral-domain optical coherence tomography (SD-OCT) is useful for visualizing retinal and ocular structures in vivo. In research, SD-OCT is a valuable ...

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