Subcellular-resolution molecular imaging within living tissue by fiber microendoscopy
Subcellular-resolution molecular imaging within living tissue by fiber microendoscopy
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DOI：
10.1364/oe.15.016413
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发表时间：
2007-12-10
期刊：
影响因子：
3.8
通讯作者：
Richards-Kortum, Rebecca
中科院分区：
文献类型：
--
作者：
Muldoon, Timothy J.;Pierce, Mark C.;Richards-Kortum, Rebecca
传统的组织病理学包括在显微镜评价之前对组织标本进行取样、切片和染色，并在单个位置和时间点提供诊断信息。体内显微镜和分子靶向光学标记是两个快速发展的领域，它们共同具有通过原位染色和成像组织来提供疾病的解剖和功能指示的潜力。为了满足高分辨率成像仪器的需求，我们开发了一种紧凑，坚固，廉价的光纤显微内窥镜系统，该系统基于宽视场LED照明，直径为1 mm的柔性光纤束和彩色CCD摄像机。我们通过一系列实验证明了该系统的亚细胞分辨率成像能力，首先对培养中的三种不同癌细胞系进行同时成像，每种细胞系都有不同的荧光标记。我们使用窄直径探针进入体内小鼠模型中的皮下肿瘤，允许直接比较显微内窥镜图像与宏观图像和组织病理学。对来自人口腔的手术切除的组织标本在临床边缘上进行成像，基于亚细胞图像特征证明正常组织和癌组织之间的定性和定量区别。最后，将光纤显微内窥镜用于局部染色的正常人口腔粘膜，在实时荧光图像中解析上皮细胞核和膜。我们的研究结果表明，这种成像系统可以潜在地补充传统的诊断技术，并支持将新兴的分子诊断和治疗剂转化为临床应用的努力。(c)2007年，美国光学学会。
Conventional histopathology involves sampling, sectioning and staining of tissue specimens prior to microscopic evaluation, and provides diagnostic information at a single location and point in time. In vivo microscopy and molecular-targeted optical labeling are two rapidly developing fields, which together have the potential to provide anatomical and functional indications of disease by staining and imaging tissue in situ. To address the need for high-resolution imaging instrumentation, we have developed a compact, robust, and inexpensive fiber-optic microendoscopy system based around wide-field LED illumination, a flexible 1 mm diameter fiber-optic bundle, and a color CCD camera. We demonstrate the subcellular resolution imaging capabilities of the system through a series of experiments, beginning with simultaneous imaging of three different cancer cell lines in culture, each targeted with a distinct fluorescent label. We used the narrow diameter probe to access subcutaneous tumors in an in vivo murine model, allowing direct comparison of microendoscopy images with macroscopic images and histopathology. A surgically resected tissue specimen from the human oral cavity was imaged across the clinical margin, demonstrating qualitative and quantitative distinction between normal and cancerous tissue based on sub-cellular image features. Finally, the fiber-optic microendoscope was used on topically-stained normal human oral mucosa in vivo, resolving epithelial cell nuclei and membranes in real-time fluorescence images. Our results demonstrate that this imaging system can potentially complement conventional diagnostic techniques, and support efforts to translate emerging molecular-diagnostic and therapeutic agents into clinical use. (c) 2007 Optical Society of America.