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首页> 外文期刊>Journal of Microscopy >Dual-mode reflectance and fluorescence near-video-rate confocal microscope for architectural, morphological and molecular imaging of tissue
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Dual-mode reflectance and fluorescence near-video-rate confocal microscope for architectural, morphological and molecular imaging of tissue

机译：双模式反射和荧光近视频共聚焦显微镜，用于组织的结构，形态和分子成像

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We have developed a near-video-rate dual-mode reflectance and fluorescence confocal microscope for the purpose of imaging ex vivo human specimens and in vivo animal models. The dual-mode confocal microscope (DCM) has light sources at 488, 664 and 784 nm, a frame rate of 15 frames per second, a maximum field of view of 300 x 250 mu m and a resolution limit of 0.31 mu m laterally and 1.37 mu m axially. The DCM can image tissue architecture and cellular morphology, as well as molecular properties of tissue, using reflective and fluorescent molecular-specific optical contrast agents. Images acquired with the DCM demonstrate that the system has the sub-cellular resolution needed to visualize the morphological and molecular changes associated with cancer progression and has the capability to image animal models of disease in vivo. In the hamster cheek pouch model of oral carcinogenesis, the DCM was used to image the epithelium and stroma of the cheek pouch; blood flow was visible and areas of dysplasia could be distinguished from normal epithelium using 6% acetic acid contrast. In human oral cavity tissue slices, DCM reflectance images showed an increase in the nuclear-to-cytoplasmic ratio and density of nuclei in neoplastic tissues as compared to normal tissue. After labelling tissue slices with fluorescent contrast agents targeting the epidermal growth factor receptor, an increase in epidermal growth factor receptor expression was detected in cancerous tissue as compared to normal tissue. The combination of reflectance and fluorescence imaging in a single system allowed imaging of two different parameters involved in neoplastic progression, providing information about both the morphological and molecular expression changes that occur with cancer progression. The dual-mode imaging capabilities of the DCM allow investigation of both morphological changes as well as molecular changes that occur in disease processes. Analyzing both factors simultaneously may be advantageous when trying to detect and diagnose disease. The DCM's high resolution and near-video-rate image acquisition and the growing inventory of molecular-specific contrast agents and disease-specific molecular markers holds significant promise for in vivo studies of disease processes such as carcinogenesis.
机译：我们已经开发了一种近视频速率双模反射和荧光共聚焦显微镜，用于对离体人体标本和体内动物模型进行成像。双模共焦显微镜（DCM）具有488、664和784 nm的光源，每秒15帧的帧速率，最大视野为300 x 250μm，横向分辨率极限为0.31μm。轴向1.37微米DCM可以使用反射和荧光分子特异性光学对比剂对组织结构和细胞形态以及组织的分子特性成像。用DCM采集的图像表明，该系统具有可视化与癌症进展相关的形态和分子变化所需的亚细胞分辨率，并具有对体内疾病动物模型进行成像的能力。在口腔癌变的仓鼠脸颊袋模型中，DCM用于对脸颊袋的上皮和间质进行成像。可见血流，使用6％的乙酸对比可将异常增生区域与正常上皮区分开。在人口腔组织切片中，DCM反射图像显示，与正常组织相比，肿瘤组织的核质比和核密度增加。用靶向表皮生长因子受体的荧光造影剂标记组织切片后，与正常组织相比，在癌组织中检测到表皮生长因子受体的表达增加。在单个系统中将反射率和荧光成像相结合，可以对肿瘤进展中涉及的两个不同参数进行成像，从而提供有关癌症进展中发生的形态和分子表达变化的信息。 DCM的双模式成像功能允许研究疾病过程中发生的形态变化以及分子变化。在尝试检测和诊断疾病时，同时分析这两个因素可能是有利的。 DCM的高分辨率和接近视频速率的图像采集，以及分子特异性造影剂和疾病特异性分子标记物的不断增加的存货，对疾病过程（如致癌作用）的体内研究具有重大前景。

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