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. 2010 Apr;90(4):521-7.
doi: 10.1016/j.exer.2010.01.008. Epub 2010 Jan 28.

Spectral domain optical coherence tomography in a murine retinal detachment model

Colleen M Cebulla  1 , Marco Ruggeri, Timothy G Murray, William J Feuer, Eleut Hernandez
Affiliations

Spectral domain optical coherence tomography in a murine retinal detachment model

Colleen M Cebulla et al. Exp Eye Res. 2010 Apr.

Abstract

Spectral domain optical coherence tomography (SD-OCT) was used to image retinal detachments in vivo, in a murine model of retinal detachment (RD). Subretinal injections of hyaluronic acid (Healon) were delivered to the right eye of seventeen 10-20 week-old C57Bl6 mice. Evaluation of the fundus with an operating microscope and fundus photography were performed. In vivo, non-contact, ultra high resolution SD-OCT imaging was performed on day 0, day 1-2, day 5-6 and day 15-16. The retinal morphology at the edge and in the area of maximal RD was evaluated. Eyes were enucleated for histologic analysis. The retinal detachment was confirmed by microscopy in all mice. The extent of the retinal detachment was evaluated by measuring the height of the retinal detachment. The retinal layers, including the photoreceptor layer, were evaluated. Retinal layers appeared indistinct soon after RD (day 1, 5), particularly over areas of maximal detachment. By day 5 and 15 the external limiting membrane was no longer visible and there was increased reflectivity of the photoreceptor layer and undulation of the outer retina in areas of RD on both SD-OCT and histology. The thickness of the outer nuclear layer and photoreceptor outer segments decreased on day 5 and 15. SD-OCT is a promising technology to follow retinal detachment and outer retinal abnormalities in a murine model.

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Figures

Figure 1
Figure 1
RDs were confirmed by microscopy in all mice. Representative fundus photographs of a mouse with no RD (A) and day 0 RD (E) are shown. H&E sections are shown with representative histopathology of RDs 1, 5, and 16 days duration (B–D,×40 magnification). Corresponding SD-OCT images taken prior to the enucleation of each animal are shown (day 1 (F, G), day5 (H, I), day 16 (J, K)). Areas corresponding to the edge (F, H, J) and maximal height ( G, I, K) of the RD at each timepoint are shown. Retinal layers appeared indistinct soon after RD (day 1 (B, F, G)), particularly over areas of maximal detachment (G). Detached areas are denoted with brackets. The detached retina at day 16 (D, K) appears thinner than day 1 (B, G) or day 5 retina (C, I).
Figure 2
Figure 2
Maximal RD height decreases over time. Average maximal height of separation of the retina from the retinal pigment epithelium in the area of detachment measured by SD-OCT is graphically displayed (n=6 day 1–2, n=3 day 5–6, n=3 day 15–16; error bars represent SD).
Figure 3
Figure 3
ELM and photoreceptor IS/OS changes on SD-OCT. In attached areas, outer retinal structures of the mouse retina are well visualized on histology (A, Magnification × 200) and the corresponding SD-OCT (B). Layers are labeled: Nerve fiber layer (NFL), ganglion cell layer (GCL), inner plexiform layer (IPL), inner nuclear layer (INL), outer plexiform layer (OPL), outer nuclear layer (ONL), external limiting membrane (ELM), photoreceptor inner segment (IS) outer segment (OS), retinal pigment epithelium (RPE). In the area of RD, the ELM in not visible, the IS and OS layers are indistinct and there is increased hyper-reflectivity of this region on SD-OCT (C, representative animal day 15). The bracket labels the subretinal space. Note that the nuclear layers are dark on SD-OCT.
Figure 4
Figure 4
Undulation of the outer retina. Representative examples of photoreceptor and outer nuclear layer undulation, undetectable ELM and outer segment degeneration seen at day 5 (A, H&E histology at × 40 magnification (a black box denotes the area shown in B at × 200 magnification) and C, the SD-OCT image of the same animal). Brackets denote the subretinal space in areas of detachment. White arrows denote points of retinal undulation and short, black arrows denote photoreceptor outer segment degeneration. Similar results are seen at day 15 (D, representative histology × 40 magnification, E, × 200 magnification of the boxed area from D, and F, SD-OCT image).
Figure 5
Figure 5
Retinal thickness and outer retinal thickness decreases with time after detachment. Total retinal thickness was quantitated by H&E histology (A) and SD-OCT (B). Changes in retinal structure thickness were determined for inner retina (Inner Ret), ONL, and PR segments by histology (C) and SD-OCT (D). Error bars represent SD. Brackets label statistically significant differences between groups: * Represents p≤0.05 and ** represents p≤0.01.
Figure 6
Figure 6
Immune infiltrate present in experimental RD. An immune infiltrate (arrows) is present in the subretinal space and photoreceptor layer in day 5–6 and day 15–16 samples (A, ×100 magnification from representative day 16 eye). Correlation of the histology (B, ×400 magnification) and SD-OCT (C) from a representative day 5 eye shows the undulation of the ONL, undetectable ELM, and high reflective structures (thick arrows) morphologically similar to, but larger than immune cells seen on histology (thin arrows). Brackets label the subretinal space.
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