Abstract
.The development of technologies to characterize the ocular aqueous outflow system (AOS) is important for the understanding of the pathophysiology of glaucoma. Multiphoton microscopy (MPM) offers the advantage of high-resolution, label-free imaging with intrinsic image contrast because the emitted signals result from the specific biomolecular content of the tissue. Previous attempts to use MPM to image the murine irido-corneal region directly through the sclera have suffered from degradation in image resolution due to scattering of the focused laser light. As a result, transscleral MPM has limited ability to observe fine structures in the AOS. In this work, the porcine irido-corneal angle was successfully imaged through the transparent cornea using a gonioscopic lens to circumvent the highly scattering scleral tissue. The resulting high-resolution images allowed the detailed structures in the trabecular meshwork (TM) to be observed. Multimodal imaging by two-photon autofluorescence and second harmonic generation allowed visualization of different features in the TM without labels and without disruption of the TM or surrounding tissues. MPM gonioscopy is a promising noninvasive imaging tool for high-resolution studies of the AOS, and research continues to explore the potential for future clinical applications in humans.
Highlights
In glaucoma, abnormalities in the ocular aqueous outflow system (AOS) can lead to elevated intraocular pressure (IOP) with subsequent death of retinal ganglion cells, resulting in loss of vision.[1]
Advancement in three-dimensional (3-D) micro– computed tomography using a synchrotron radiation source can achieve 2-μm voxel resolution, which is adequate for resolving details of the AOS structures including trabecular meshwork (TM), s canal (SC), and collector channels (CC).[11]
We explore the use of a unique Multiphoton microscopy (MPM) setup that leverages a gonioscopic lens for direct transcorneal imaging of the AOS in an intact porcine eye
Summary
Abnormalities in the ocular aqueous outflow system (AOS) can lead to elevated intraocular pressure (IOP) with subsequent death of retinal ganglion cells, resulting in loss of vision.[1]. Current diagnostic techniques used to study the AOS include gonioscopy,[2] optical coherence tomography (OCT),[3,4,5,6,7,8] and ultrasound biomicroscopy (UBM).[9,10] None of these techniques can image with molecular specificity, nor do they have the spatial resolution (∼1 to 5 μm) required to resolve TM structures. Advancement in three-dimensional (3-D) micro– computed tomography (micro-CT) using a synchrotron radiation source can achieve 2-μm voxel resolution, which is adequate for resolving details of the AOS structures including TM, SC, and CC.[11] beam line availability and the associated costs of synchrotron radiation sources prohibit the use of this technique in most laboratories, and the required use of contrast agents prevents its routine clinical use for studying the ocular AOS
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