Greening Gage J, Powless Amy J, Hutcheson Joshua A, Prieto Sandra P, Majid Aneeka A, Muldoon Timothy J
Department of Biomedical Engineering, University of Arkansas, Fayetteville, AR, USA 72701.
Proc SPIE Int Soc Opt Eng. 2015 Mar 9;9332:93320R. doi: 10.1117/12.2076300.
Many cases of epithelial cancer originate in basal layers of tissue and are initially undetected by conventional microendoscopy techniques. We present a bench-top, fiber-bundle microendoscope capable of providing high resolution images of surface cell morphology. Additionally, the microendoscope has the capability to interrogate deeper into material by using diffuse reflectance and broadband diffuse reflectance spectroscopy. The purpose of this multimodal technique was to overcome the limitation of microendoscopy techniques that are limited to only visualizing morphology at the tissue or cellular level. Using a custom fiber optic probe, high resolution surface images were acquired using topical proflavine to fluorescently stain non-keratinized epithelia. A 635 nm laser coupled to a 200 μm multimode fiber delivers light to the sample and the diffuse reflectance signal was captured by a 1 mm image guide fiber. Finally, a tungsten-halogen lamp coupled to a 200 μm multimode fiber delivers broadband light to the sample to acquire spectra at source-detector separations of 374, 729, and 1051 μm. To test the instrumentation, a high resolution proflavine-induced fluorescent image of resected healthy mouse colon was acquired. Additionally, five monolayer poly(dimethylsiloxane)-based optical phantoms with varying absorption and scattering properties were created to acquire diffuse reflectance profiles and broadband spectra.
许多上皮癌病例起源于组织的基底层，最初无法通过传统的显微内窥镜技术检测到。我们展示了一种台式纤维束显微内窥镜，它能够提供表面细胞形态的高分辨率图像。此外，该显微内窥镜还能够通过使用漫反射和宽带漫反射光谱对更深层的物质进行检测。这种多模态技术的目的是克服显微内窥镜技术的局限性，即仅限于在组织或细胞水平观察形态。使用定制的光纤探头，通过局部使用硫酸原黄素对非角化上皮进行荧光染色来获取高分辨率表面图像。一台与200μm多模光纤耦合的635nm激光器将光传输到样品，漫反射信号由一根1mm的图像引导光纤捕获。最后，一台与200μm多模光纤耦合的钨卤灯将宽带光传输到样品，以在源探测器间距为374、729和1051μm时获取光谱。为了测试该仪器，获取了切除的健康小鼠结肠的高分辨率硫酸原黄素诱导荧光图像。此外，制作了五个具有不同吸收和散射特性的基于聚二甲基硅氧烷的单层光学模型，以获取漫反射轮廓和宽带光谱。
