The modern otoscope is an indispensable instrument utilized by primary care physicians as the gold standard tool to diagnose an array of otologic diseases and conditions. At present, commercially available, traditional otoscopes remain cost-prohibitive to many potential users despite limited innovation since its invention in the early 19th century. In this publication, the design and assembly of a low-cost, open-source, 3D-printed otoscope, the Glia Otoscope V1.0, is outlined. Subsequently, we describe the benchtop evaluation conducted, which measured several outcomes relevant to otoscopy performance against a traditional, gold standard otoscope, the Welch Allyn Rechargeable V3.5 Halogen HPX Otoscope. Measured outcomes included illuminance, correlated color temperature, color rendering index, spatial resolution, field of view, weight, battery life, and cost. Overall, the Glia Otoscope V1.0 demonstrated comparable performance across measured outcomes against the traditional otoscope. Further validation in the clinical setting is warranted as the Glia Otoscope V1.0 and its future iterations hold tremendous potential in improving access and alleviating the burden of otologic disease in lower and middle-income countries. Finally, we present a novel tool, the Otoscope Assessment Tool, which establishes a standard set of performance characteristics for benchtop evaluation of otoscope performance.