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1.
Invest Ophthalmol Vis Sci ; 59(5): 2054-2068, 2018 04 01.
Article in English | MEDLINE | ID: mdl-29677368

ABSTRACT

Purpose: To investigate the early development of spontaneous retinal neovascularization in the murine retina by a multifunctional optical coherence tomography approach. To characterize involved tissue changes in vivo and describe structural and functional changes over time. Methods: A multifunctional optical coherence tomography (OCT) system providing 3-fold contrast comprising reflectivity, polarization sensitivity, and OCT angiography (OCTA) was utilized to image very-low-density lipoprotein receptor (VLDLR) knockout mice. Baseline measurements were acquired as early as postnatal day 14 and a follow-up of neovascularization development was performed until the age of 3 months. Control mice were imaged accordingly and a multiparametric image analysis was performed to characterize different stages of pathologic vascular growth. Histology was conducted at the endpoint of the experiment. An interventional pilot experiment was conducted to investigate the effect of the anti-vascular endothelial growth factor (VEGF) agent aflibercept on the development of retinal neovascularization. Results: Onset of neovascularization was imaged at baseline, and significant changes were encountered in the retina over time, including reduced retinal thickness, increase of lesion volume, migration of pigmented structures, and presence of abnormal blood flow in the outer retina. Multifunctional image contrast was correlated to ex vivo histology. Microscopic analysis of retinal flat mounts and cross-sectional samples confirmed the changes observed in in vivo structural and functional OCT images. Administration of an anti-VEGF agent resulted in a significantly reduced lesion volume. Conclusions: Longitudinal, multifunctional OCT imaging of infant VLDLR-/- mouse retinas enabled a multiparametric, in vivo staging of neovascularization formation from before lesion onset until their manifestation.


Subject(s)
Retinal Neovascularization/diagnostic imaging , Retinal Vessels/diagnostic imaging , Tomography, Optical Coherence/methods , Animals , Cross-Sectional Studies , Female , Fluorescein Angiography , Male , Mice , Mice, Knockout , Multimodal Imaging , Receptors, LDL/genetics
2.
Sci Rep ; 7(1): 8780, 2017 08 18.
Article in English | MEDLINE | ID: mdl-28821834

ABSTRACT

A multi-functional optical coherence tomography (OCT) approach is presented to determine ocular fundus pulsations as an axial displacement between the retina and the chorioscleral complex in the albino rat eye. By combining optical coherence elastography and OCT angiography (OCTA), we measure subtle deformations in the nanometer range within the eye and simultaneously map retinal and choroidal perfusion. The conventional OCT reflectivity contrast serves as a backbone to segment the retina and to define several slabs which are subsequently used for quantitative ocular pulsation measurements as well as for a qualitative exploration of the multi-functional OCT image data. The proposed concept is applied in healthy albino rats as well as in rats under acute elevation of the intraocular pressure (IOP). The evaluation of this experiment revealed an increased pulsatility and deformation between the retinal and chorioscleral complex while increasing the IOP level from 15 mmHg to 65 mmHg. At IOP levels exceeding 65 mmHg, the pulsatility decreased significantly and retinal as well as choroidal perfusion vanished in OCTA. Furthermore, the evaluation of the multi-parametric experiment revealed a spatial correlation between fundus pulsatility and choroidal blood flow. This indicates that the assessed pulsatility may be a valuable parameter describing the choroidal perfusion.


Subject(s)
Choroid/blood supply , Fundus Oculi , Intraocular Pressure , Angiography , Animals , Male , Perfusion , Rats , Reproducibility of Results , Tomography, Optical Coherence
3.
Biomed Opt Express ; 8(1): 298-314, 2017 Jan 01.
Article in English | MEDLINE | ID: mdl-28101419

ABSTRACT

Polarization sensitive optical coherence tomography (PS-OCT) operating at 840 nm with axial resolution of 3.8 µm in tissue was used for investigating the posterior rat eye during an acute intraocular pressure (IOP) increase experiment. IOP was elevated in the eyes of anesthetized Sprague Dawley rats by cannulation of the anterior chamber. Three dimensional PS-OCT data sets were acquired at IOP levels between 14 mmHg and 105 mmHg. Maps of scleral birefringence, retinal nerve fiber layer (RNFL) retardation and relative RNFL/retina reflectivity were generated in the peripapillary area and quantitatively analyzed. All investigated parameters showed a substantial correlation with IOP. In the low IOP range of 14-45 mmHg only scleral birefringence showed statistically significant correlation. The polarization changes observed in the PS-OCT imaging study presented in this work suggest that birefringence of the sclera may be a promising IOP-related parameter to investigate.

4.
PLoS One ; 11(10): e0164419, 2016.
Article in English | MEDLINE | ID: mdl-27711217

ABSTRACT

We present a multi-functional optical coherence tomography (OCT) imaging approach to study retinal changes in the very-low-density-lipoprotein-receptor (VLDLR) knockout mouse model with a threefold contrast. In the retinas of VLDLR knockout mice spontaneous retinal-chorodoidal neovascularizations form, having an appearance similar to choroidal and retinal neovascularizations (CNV and RNV) in neovascular age-related macular degeneration (AMD) or retinal angiomatous proliferation (RAP). For this longitudinal study, the mice were imaged every 4 to 6 weeks starting with an age of 4 weeks and following up to the age of 11 months. Significant retinal changes were identified by the multi-functional imaging approach offering a threefold contrast: reflectivity, polarization sensitivity (PS) and motion contrast based OCT angiography (OCTA). By use of this intrinsic contrast, the long-term development of neovascularizations was studied and associated processes, such as the migration of melanin pigments or retinal-choroidal anastomosis, were assessed in vivo. Furthermore, the in vivo imaging results were validated with histological sections at the endpoint of the experiment. Multi-functional OCT proves as a powerful tool for longitudinal retinal studies in preclinical research of ophthalmic diseases. Intrinsic contrast offered by the functional extensions of OCT might help to describe regulative processes in genetic animal models and potentially deepen the understanding of the pathogenesis of retinal diseases such as wet AMD.


Subject(s)
Receptors, LDL/genetics , Retina/diagnostic imaging , Animals , Choroidal Neovascularization/diagnostic imaging , Choroidal Neovascularization/pathology , Image Processing, Computer-Assisted , Longitudinal Studies , Mice , Mice, Inbred C57BL , Mice, Knockout , Receptors, LDL/deficiency , Retina/pathology , Retina/physiology , Retinal Neovascularization/diagnostic imaging , Retinal Neovascularization/pathology , Tomography, Optical Coherence
5.
Biomed Opt Express ; 7(4): 1479-95, 2016 Apr 01.
Article in English | MEDLINE | ID: mdl-27446670

ABSTRACT

We present a high resolution polarization sensitive optical coherence tomography (PS-OCT) system for ocular imaging in rodents. The system operates at 840 nm and uses a broadband superluminescent diode providing an axial resolution of 5.1 µm in air. PS-OCT data was acquired at 83 kHz A-scan rate by two identical custom-made spectrometers for orthogonal polarization states. Pigmented (Brown Norway, Long Evans) and non-pigmented (Sprague Dawley) rats as well as pigmented mice (C57BL/6) were imaged. Melanin pigment related depolarization was analyzed in the retinal pigment epithelium (RPE) and choroid of these animals using the degree of polarization uniformity (DOPU). For all rat strains, significant differences between RPE and choroidal depolarization were observed. In contrast, DOPU characteristics of RPE and choroid were similar for C57BL/6 mice. Moreover, the depolarization within the same tissue type varied significantly between different rodent strains. Retinal nerve fiber layer thickness, phase retardation, and birefringence were mapped and quantitatively measured in Long Evans rats in vivo for the first time. In a circumpapillary annulus, retinal nerve fiber layer birefringence amounted to 0.16°/µm ± 0.02°/µm and 0.17°/µm ± 0.01°/µm for the left and right eyes, respectively.

6.
Invest Ophthalmol Vis Sci ; 56(12): 7462-72, 2015 Nov.
Article in English | MEDLINE | ID: mdl-26595606

ABSTRACT

PURPOSE: The purpose of this study was to demonstrate polarization-sensitive optical coherence tomography (PS-OCT) for imaging pigmented structures in the posterior eye segments of albino and pigmented rats and to correlate depolarization contrast of the retinal pigment epithelium (RPE) and choroid in in vivo PS-OCT to melanin pigmentation detected in postmortem histologic serial sections. METHODS: In vivo three-dimensional PS-OCT imaging was performed in adult albino and pigmented rat eyes at 70-kHz A-line rate. Degree of polarization uniformity (DOPU) fundus maps and radial DOPU profiles were generated. Postmortem histomorphologic analysis was performed in order to investigate melanin pigmentation of the RPE and choroid. Fundus pigmentation maps were extracted from histologic serial sections. Pigmentation profiles were correlated to DOPU profiles of the same eyes. RESULTS: Strong depolarization was found in the RPE/choroid complex of pigmented rats, whereas the same structures exhibited uniform polarization in albino rats. The difference between the depolarization characteristics between albino and pigmented animals was statistically significant. In the fundus pigmentation maps, optical pigment density was zero in albino rat eyes. In pigmented rat eyes, a strong negative correlation between optical pigment density and DOPU was observed. CONCLUSIONS: This in vivo and ex vivo investigation of posterior rat eyes indicates that melanin is the cause of depolarization in retinal PS-OCT images. It further demonstrates that melanin pigmentation in the RPE and choroid can be quantified via depolarization imaging and therefore suggests that PS-OCT is a useful tool for the noninvasive quantitative assessment of pigmentary changes in vision-threatening diseases such as age-related macular degeneration.


Subject(s)
Albinism/pathology , Melanins/metabolism , Retinal Pigment Epithelium/pathology , Tomography, Optical Coherence/methods , Animals , Disease Models, Animal , Fluorescein Angiography , Fundus Oculi , Male , Rats , Rats, Inbred BN , Rats, Long-Evans , Rats, Sprague-Dawley , Retinal Pigment Epithelium/metabolism , Visual Acuity
7.
Invest Ophthalmol Vis Sci ; 55(11): 7686-96, 2014 Oct 28.
Article in English | MEDLINE | ID: mdl-25352116

ABSTRACT

PURPOSE: To demonstrate polarization-sensitive (PS) optical coherence tomography (OCT) for noninvasive, volumetric, and quantitative imaging of the birefringent properties of the peripapillary rat sclera; to compare the findings from PS-OCT images to state-of-the-art histomorphometric analysis of the same tissues. METHODS: A high-speed PS-OCT prototype operating at 840 nm was modified for imaging the rat eye. Densely sampled PS-OCT raster scans covering an area of ~1.5 × 1.5 mm centered at the papilla were acquired in the eyes of anesthetized male Sprague-Dawley rats. Cross-sectional PS-OCT images were computed, and fundus maps displaying the birefringent properties of the sclera were analyzed. Postmortem histomorphologic analysis was performed. RESULTS: Polarization-sensitive OCT enables visualization of the polarization properties of ocular tissues in vivo. The birefringent characteristics of the rat sclera were quantitatively assessed. Scleral birefringence formed a donut-shaped pattern around the papilla with significantly increased values of 0.703 ± 0.089°/µm (i.e., 1.64 × 10(-3) ± 0.2 × 10(-3); mean ± standard deviation) and 0.721 ± 0.084°/µm (i.e., 1.68 × 10(-3) ± 0.2 × 10(-3)) at an eccentricity of 0.4 mm for the left and right eyes, respectively. Birefringent axis orientation maps revealed a ring-shaped distribution around the optic nerve. Postmortem PS-OCT micrographs provided access to retinal and scleral microstructure and were compared to standard histomorphologic analysis. CONCLUSIONS: Polarization-sensitive OCT enables quantitative imaging of tissue polarization properties in addition to conventional OCT imaging based on reflectivity. In the rat sclera, in vivo PS-OCT provides access to volumetric mapping of birefringence. Scleral birefringence is associated with microstructural tissue organization. Therefore, PS-OCT should prove a valuable tool for the in vivo investigation of peripapillary sclera in glaucoma.


Subject(s)
Sclera/cytology , Tomography, Optical Coherence/instrumentation , Animals , Birefringence , Equipment Design , Male , Rats , Rats, Sprague-Dawley , Reproducibility of Results
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