Multimodal photoacoustic remote sensing (PARS) microscopy combined with swept-source optical coherence tomography (SS-OCT) for in-vivo, non-contact, functional and structural ophthalmic imaging applications

Ophthalmic imaging has long played an important role in the understanding, diagnosis, and treatment of a wide variety of ocular disorders. Currently, available clinical ophthalmic imaging instruments are primarily optical-based, including slit-lamp microscopy, fundus photography, confocal microscopy, scanning laser ophthalmoscopy, and optical coherence tomography (OCT). The development of these imaging instruments has greatly extended our ability to evaluate the ocular environment. Studies have shown that at least 40% of blinding disorders in the United States are either preventable or treatable with timely diagnosis and intervention. OCT is a state-of-the-art imaging technique extensively used in preclinical and clinical applications for imaging both anterior and posterior parts of the eye. OCT has become a standard of care for the assessment and treatment of most ocular conditions. The technology enables non-contact, high-speed, cross-sectional imaging over a large field of view with submicron resolutions. In eye imaging applications, functional extensions of OCT such as spectroscopic OCT and Doppler OCT have been applied to provide a better understanding of tissue activity. Spectroscopic OCT is usually achieved through OCT systems in the visible spectral range, and it enables the amount of light absorption inside the ocular environment to be measured. This indirect optical absorption measurement is used to estimate the amount of ocular oxygen saturation (SO2) which is a well-known biomarker in prevalent eye diseases including diabetic retinopathy, glaucoma, and retinal vein occlusions. Despite all the advancements in functional spectroscopic OCT methods, they still rely primarily on measuring the backscattered photons to quantify the absorption of chromophores inside the tissue. Therefore, they are sensitive to local geometrical parameters, such as retinal thickness, vessel diameters, and retinal pigmentation, and may result in biased estimations. Of the various optical imaging modalities, photoacoustic imaging (PAI) offers unique imaging contrast of optical absorption because PAI can image any target that absorbs light energy. This unique imaging ability makes PAI a favorable candidate for various functional and molecular imaging applications as well as for measuring chromophore concentration. Over the past decade, photoacoustic ophthalmoscopy has been applied for visualizing hemoglobin and melanin content in ocular tissue, quantifying ocular SO2, and measuring the metabolic rate of oxygen consumption (MRO2). Despite all these advantages offered by PAI devices, a major limitation arises from their need to be in contact with the ocular tissues. This physical contact may increase the risk of infection and cause patient discomfort. Furthermore, this contact-based imaging approach applies pressure to the eye and introduces barriers to oxygen diffusion. Thus, it has a crucial influence on the physiological and pathophysiological balance of ocular vasculature function, and it is not capable of studying dynamic processes under normal conditions. To overcome these limitations and to benefit from the numerous advantages offered by photoacoustic ophthalmoscopy, non-contact detection of photoacoustic signals has been a long-lasting goal in the field of ocular imaging. In 2017 Haji Reza et al. developed photoacoustic remote sensing (PARS) for non-contact, non-interferometric detection of photoacoustic signals. PARS is the non-contact, all-optical version of optical-resolution photoacoustic microscopy (OR-PAM), where the acoustically coupled ultrasound transducer is replaced with a co-focused probe beam. This all-optical detection scheme allows the system to measure the photoacoustic pressure waves at the subsurface origin where the pressure is at a maximum. In a very short time, PARS technology has proven its potential for various biomedical applications, including label-free histological imaging, SO2 mapping, and angiogenesis imaging. PARS is an ideal companion for OCT in ophthalmic applications, where the depth-resolved, detailed scattering information of OCT is well complemented by rich absorption information of PARS. This combined multimodal imaging technology has the potential to provide chromophore selective absorption contrast in concert with depth-resolved scattering contrast in the ocular environment. The main goals of this PhD project are to: • Develop a photoacoustic remote sensing microscopy system for in-vivo, non-contact ophthalmic imaging. This is the first time a non-contact photoacoustic imaging has been used for in-vivo imaging of the eye. • Develop a robust and temporally stable multiwavelength light source for functional photoacoustic imaging applications. • Develop a multimodal PARS-OCT imaging system that can image in-vivo and record, simultaneously, functional, and structural information in the anterior segment of a rodent eye. This is the first time a multiwavelength non-contact photoacoustic system is used for in-vivo measurement of oxygen saturation in the ocular environment. • Develop and modify the multimodal PARS-OCT imaging system for non-contact, in-vivo, functional, and structural imaging of the posterior part of the rodent eye

Comparison of phase-resolved Doppler optical coherence tomography and optical coherence tomography angiography for measuring retinal blood vessels size

The goal of this study was to compare two OCT-based methods for measuring retinal blood vessels size: Phase-resolved Doppler OCT (DOCT) and OCT angiography (OCTA). The study was conducted in rats (n= 6) using a SD-OCT system operating at 1060 nm with 92 kHz image acquisition rate. Arteries and veins were separated by the phase polarity. Results from this study showed that the venal diameters are significantly larger than the arterial diameters, and there is no significant difference in the vessel diameters measured by both methods

Line-scanning SD-OCT for in-vivo, non-contact, volumetric, cellular resolution imaging of the human cornea and limbus

In-vivo, non-contact, volumetric imaging of the cellular and sub-cellular structure of the human cornea and limbus with optical coherence tomography (OCT) is challenging due to involuntary eye motion that introduces both motion artifacts and blur in the OCT images. Here we present the design of a line-scanning (LS) spectral-domain (SD) optical coherence tomography system that combines 2 × 3 × 1.7 µm (x, y, z) resolution in biological tissue with an image acquisition rate of ∼2,500 fps, and demonstrate its ability to image in-vivo and without contact with the tissue surface, the cellular structure of the human anterior segment tissues. Volumetric LS-SD-OCT images acquired over a field-of-view (FOV) of 0.7 mm × 1.4 mm reveal fine morphological details in the healthy human cornea, such as epithelial and endothelial cells, sub-basal nerves, as well as the cellular structure of the limbal crypts, the palisades of Vogt (POVs) and the blood microvasculature of the human limbus. LS-SD-OCT is a promising technology that can assist ophthalmologists with the early diagnostics and optimal treatment planning of ocular diseases affecting the human anterior eye.Published versionCanada First Research Excellence Fund; Canadian Institutes of Health Research (446387); Natural Sciences and Engineering Research Council of Canada (312037)