Dual-mode reflectance and fluorescence near-video-rate confocal microscope for architectural, morphological and molecular imaging of tissue
Dual-mode reflectance and fluorescence near-video-rate confocal microscope for architectural, morphological and molecular imaging of tissue
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DOI：
10.1111/j.1365-2818.2007.01818.x
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发表时间：
2007-10-01
影响因子：
2
通讯作者：
Richards-Kortum, Rebecca R.
中科院分区：
文献类型：
--
作者：
Carlson, Alicia L.;Coghlan, Lezlee G.;Richards-Kortum, Rebecca R.
我们已经开发了一个近视频率的双模反射和荧光共聚焦显微镜成像离体人体标本和在体动物模型的目的。双模式共聚焦显微镜（DCM）具有488，664和784 nm的光源，每秒15帧的帧速率，300 × 250 μ m的最大视场和0.31 μ m的横向和1.37 μ m的轴向分辨率极限。DCM可以使用反射和荧光分子特异性光学造影剂对组织结构和细胞形态以及组织的分子特性进行成像，使用DCM获得的图像表明，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，并且具有对体内疾病动物模型进行成像的能力。在口腔癌发生的仓鼠颊囊模型中，DCM用于对颊囊的上皮和基质成像;血流可见，并且使用6%乙酸造影剂可以将发育异常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，肿瘤组织中的核与细胞质比率和核密度增加。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表达的增加。在一个单一的系统中的反射和荧光成像的组合允许成像的两个不同的参数参与肿瘤的进展，提供信息的形态和分子表达的变化，发生与cancer progression.The双模式成像能力的DCM允许调查的形态变化，以及发生在疾病过程中的分子变化。当试图检测和诊断疾病时，同时分析这两个因素可能是有利的。DCM的高分辨率和近视频速率的图像采集和不断增长的库存的分子特异性造影剂和疾病特异性分子标记物持有显着的承诺，在体内研究的疾病过程，如致癌作用。
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.