On-axis full-field swept-source optical coherence tomography for murine retinal imaging.

A full-field swept-source optical coherence tomography (FF-SS-OCT) for in vivo murine retinal imaging is demonstrated. The on-axis FF-SS-OCT system was built in a Mach-Zehnder interferometer configuration employing a tunable laser source with an adjustable sweep rate and sweep range in conjunction with a fast 2D-CMOS camera. A large field retinal (coherent) illumination was accomplished using an imaging interface comprised of a short-focal length imaging lens and a contact lens.

High-speed, phase contrast retinal and blood flow imaging using an adaptive optics partially confocal multi-line ophthalmoscope.

High-speed, phase contrast retinal and blood flow imaging using an adaptive optics partially confocal multi-line ophthalmosocope (AO-pcMLO) is described. It allows for simultaneous confocal and phase contrast imaging with various directional multi-line illumination by using a single 2D camera and a digital micromirror device (DMD). Both vertical and horizontal line illumination directions were tested, for photoreceptor and vascular imaging.

Programmable, high-speed, adaptive optics partially confocal multi-spot ophthalmoscope using a digital micromirror device.

A high-speed, adaptive optics partially confocal multi-spot ophthalmoscope (AO-pcMSO) using a digital micromirror device (DMD) in the illumination channel and a fast 2D CMOS camera is described. The camera is synchronized with the DMD allowing projection of multiple, simultaneous AO-corrected spots onto the human retina. Spatial filtering on each raw retinal image before reconstruction works as an array virtual pinholes.

In Vivo Imaging of Retinal and Choroidal Morphology and Vascular Plexuses of Vertebrates Using Swept-Source Optical Coherence Tomography.

Purpose: To perform in vivo evaluation of the structural morphology and vascular plexuses of the neurosensory retina and choroid across vertebrate species using swept-source optical coherence tomography (SS-OCT) and SS-OCT angiography (SS-OCTA) imaging.

Methods: A custom-built SS-OCT system with an incorporated flexible imaging arm was used to acquire the three-dimensional (3D) retinal OCT and vascular OCTA data of five different vertebrates: a mouse (C57BL/6J), a rat (Long Evans), a gray short-tailed opossum (Monodelphis domestica), a white sturgeon (Acipenser transmontanus), and a great horned owl (Bubo virginianus).

Results: In vivo structural morphology of the retina and choroid, as well as en face OCTA images of retinal and choroidal vasculature of all species were generated.

CRISPR-based VEGF suppression using paired guide RNAs for treatment of choroidal neovascularization.

Clustered regularly interspaced short palindromic repeats (CRISPR)-based genomic disruption of vascular endothelial growth factor A () with a single gRNA suppresses choroidal neovascularization (CNV) in preclinical studies, offering the prospect of long-term anti-angiogenesis therapy for neovascular age-related macular degeneration (AMD). Genome editing using CRISPR-CRISPR-associated endonucleases (Cas9) with multiple guide RNAs (gRNAs) can enhance gene-ablation efficacy by augmenting insertion-deletion (indel) mutations with gene truncations but may also increase the risk of off-target effects. In this study, we compare the effectiveness of adeno-associated virus (AAV)-mediated CRISPR-Cas9 systems using single versus paired gRNAs to target two different loci in the gene that are conserved in human, rhesus macaque, and mouse.

In vivo multimodal retinal imaging of disease-related pigmentary changes in retinal pigment epithelium.

Melanosomes, lipofuscin, and melanolipofuscin are the three principal types of pigmented granules found in retinal pigment epithelium (RPE) cells. Changes in the density of melanosomes and lipofuscin in RPE cells are considered hallmarks of various retinal diseases, including Stargardt disease and age-related macular degeneration (AMD). Herein, we report the potential of an in vivo multimodal imaging technique based on directional back-scattering and short-wavelength fundus autofluorescence (SW-FAF) to study disease-related changes in the density of melanosomes and lipofuscin granules in RPE cells.

An efficient retinal blood vessel segmentation in eye fundus images by using optimized top-hat and homomorphic filtering.

Background And Objective: Automatic segmentation of retinal blood vessels makes a major contribution in CADx of various ophthalmic and cardiovascular diseases. A procedure to segment thin and thick retinal vessels is essential for medical analysis and diagnosis of related diseases. In this article, a novel methodology for robust vessel segmentation is proposed, handling the existing challenges presented in the literature.

In Situ Morphologic and Spectral Characterization of Retinal Pigment Epithelium Organelles in Mice Using Multicolor Confocal Fluorescence Imaging.

Purpose: To investigate the major organelles of the retinal pigment epithelium (RPE) in wild-type (WT, control) mice and their changes in pigmented Abca4 knockout (Abca4-/-) mice with in situ morphologic, spatial, and spectral characterization of live ex vivo flat-mounted RPE using multicolor confocal fluorescence microscopy (MCFM).

Methods: In situ imaging of RPE flat-mounts of agouti Abca4-/- (129S4), agouti WT (129S1/SvlmJ) controls, and B6 albino mice (C57BL/6J-Tyrc-Brd) was performed with a Nikon A1 confocal microscope. High-resolution confocal image z-stacks of the RPE cell mosaic were acquired with four different excitation wavelengths (405 nm, 488 nm, 561 nm, and 640 nm).

Factors Impacting Efficacy of AAV-Mediated CRISPR-Based Genome Editing for Treatment of Choroidal Neovascularization.

Frequent injections of anti-vascular endothelial growth factor (anti-VEGF) agents are a clinical burden for patients with neovascular age-related macular degeneration (AMD). Genomic disruption of using adeno-associated viral (AAV) delivery of clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 has the potential to permanently suppress aberrant angiogenesis, but the factors that determine the optimal efficacy are unknown. Here, we investigate two widely used Cas9 endonucleases, SpCas9 and SaCas9, and evaluate the relative contribution of AAV-delivery efficiency and genome-editing rates to determine the mechanisms that drive successful CRISPR-based suppression of VEGF-A, using a mouse model of laser-induced choroidal neovascularization (CNV).

Directional optical coherence tomography reveals melanin concentration-dependent scattering properties of retinal pigment epithelium.

Optical coherence tomography (OCT) is a powerful tool in ophthalmology that provides in vivo morphology of the retinal layers and their light scattering properties. The directional (angular) reflectivity of the retinal layers was investigated with focus on the scattering from retinal pigment epithelium (RPE). The directional scattering of the RPE was studied in three mice strains with three distinct melanin concentrations: albino (BALB/c), agouti (129S1/SvlmJ), and strongly pigmented (C57BL/6J).

Aperture phase modulation with adaptive optics: a novel approach for speckle reduction and structure extraction in optical coherence tomography.

Speckle is an inevitable consequence of the use of coherent light in imaging and acts as noise that corrupts image formation in most applications. Optical coherence tomographic imaging, as a technique employing coherence time gating, suffers from speckle. We present here a novel method of suppressing speckle noise intrinsically compatible with adaptive optics (AO) for confocal coherent imaging: modulation of the phase in the system pupil aperture with a segmented deformable mirror (DM) to introduce minor perturbations in the point spread function.

Effect of a contact lens on mouse retinal in vivo imaging: Effective focal length changes and monochromatic aberrations.

For in vivo mouse retinal imaging, especially with Adaptive Optics instruments, application of a contact lens is desirable, as it allows maintenance of cornea hydration and helps to prevent cataract formation during lengthy imaging sessions. However, since the refractive elements of the eye (cornea and lens) serve as the objective for most in vivo retinal imaging systems, the use of a contact lens, even with 0 Dpt. refractive power, can alter the system's optical properties.

Quantitative optical coherence microscopy for the in situ investigation of the biofilm.

This paper explores the potential of optical coherence microscopy (OCM) for the View Article and Find Full Text PDF

Plasmon Resonant Silica-Coated Silver Nanoplates as Contrast agents for Optical Coherence Tomography.

Silica-coated silver nanostructures are identified as potential contrast agents for visible and near-infrared bio-imaging applications due to their high optical extinction caused by localized surface plasmon resonance (LSPR), improved chemical stability, and lower toxicity. We demonstrate the potential of plasmon resonant silica-coated silver nanoplates as a contrast agent for optical coherence tomography (OCT). It is shown that, triangular-shaped silica-coated silver nanoplates (SSNPs) with a side length of 170 ± 5 nm, base silver thickness of 10 ± 1 nm, and silica shell thickness of 40 ± 2 nm, exhibit higher optical extinction at a 1300 nm wavelength range, thus making them an excellent contrast agent for OCT imaging.