Purpose: There is a growing interest in targeting minimally invasive surgery devices to the aqueous outflow system to optimize treatment outcomes. However, methods to visualize functioning, large-caliber aqueous and episcleral veins in-vivo are lacking. This pilot study establishes an ex-vivo system to evaluate the use of a confocal laser microendoscope to noninvasively image episcleral vessels and quantify regional flow variation along the limbal circumference. Methods: A fiber-optic confocal laser endomicroscopy (CLE) system with lateral and axial resolution of 3.5 μ m and 15 μ m, respectively, was used on three porcine and four human eyes. Diluted fluorescein (0.04%) was injected into eyes kept under constant infusion. The microprobe was applied to the sclera 1 mm behind the limbus to acquire real-time video. Image acquisition was performed at 15-degree intervals along the limbal circumference to quantify regional flow variation in human eyes. Results: Vascular structures were visualized in whole human eyes without processing. Schlemm's canal was visualized only after a scleral flap was created. Fluorescent signal intensity and vessel diameter variation were observed along the limbal circumference, with the inferior quadrant having a statistically higher fluorescein signal compared to the other quadrants in human eyes ( P < 0.05). Conclusion: This study demonstrates for the first time that the fiber-optic CLE platform can visualize the episcleral vasculature with high resolution ex-vivo with minimal tissue manipulation. Intravascular signal intensities and vessel diameters were acquired in real-time; such information can help select target areas for minimally invasive glaucoma surgery (MIGS) to achieve greater intraocular pressure reduction.
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