Carlson Alicia L, Gillenwater Ann M, Williams Michelle D, El-Naggar Adel K, Richards-Kortum R R
Dept. of Biomedical Engineering, The University of Texas at Austin, 1 University Station CO800, Austin, TX 78712, USA.
Technol Cancer Res Treat. 2007 Oct;6(5):361-74. doi: 10.1177/153303460700600501.
Using current clinical diagnostic techniques, it is difficult to visualize tumor morphology and architecture at the cellular level, which is necessary for diagnostic localization of pathologic lesions. Optical imaging techniques have the potential to address this clinical need by providing real-time, sub-cellular resolution images. This paper describes the use of dual mode confocal microscopy and optical molecular-specific contrast agents to image tissue architecture, cellular morphology, and sub-cellular molecular features of normal and neoplastic oral tissues. Fresh tissue slices were prepared from 33 biopsies of clinically normal and abnormal oral mucosa obtained from 14 patients. Reflectance confocal images were acquired after the application of 6% acetic acid, and fluorescence confocal images were acquired after the application of a fluorescence contrast agent targeting the epidermal growth factor receptor (EGFR). The dual imaging modes provided images similar to light microscopy of hematoxylin and eosin and immunohistochemistry staining, but from thick fresh tissue slices. Reflectance images provided information on the architecture of the tissue and the cellular morphology. The nuclear-to-cytoplasmic (N/C) ratio from the reflectance images was at least 7.5 times greater for the carcinoma than the corresponding normal samples, except for one case of highly keratinized carcinoma. Separation of carcinoma from normal and mild dysplasia was achieved using this ratio (p<0.01). Fluorescence images of EGFR expression yielded a mean fluorescence labeling intensity (FLI) that was at least 2.7 times higher for severe dysplasia and carcinoma samples than for the corresponding normal sample, and could be used to distinguish carcinoma from normal and mild dysplasia (p<0.01). Analyzed together, the N/C ratio and the mean FLI may improve the ability to distinguish carcinoma from normal squamous epithelium.
使用当前的临床诊断技术，很难在细胞水平上可视化肿瘤形态和结构，而这对于病理病变的诊断定位是必要的。光学成像技术有潜力通过提供实时的亚细胞分辨率图像来满足这一临床需求。本文描述了使用双模式共聚焦显微镜和光学分子特异性造影剂对正常和肿瘤性口腔组织的组织结构、细胞形态和亚细胞分子特征进行成像。从14例患者的33份临床正常和异常口腔黏膜活检样本中制备新鲜组织切片。在应用6%醋酸后采集反射共聚焦图像，在应用靶向表皮生长因子受体（EGFR）的荧光造影剂后采集荧光共聚焦图像。这两种成像模式提供的图像类似于苏木精和伊红染色及免疫组织化学染色的光学显微镜图像，但来自较厚的新鲜组织切片。反射图像提供了有关组织结构和细胞形态的信息。除了一例高度角化的癌外，癌组织反射图像中的核质比（N/C）至少是相应正常样本的7.5倍。使用该比例可实现癌组织与正常及轻度发育异常组织的区分（p<0.01）。EGFR表达的荧光图像显示，重度发育异常和癌组织样本的平均荧光标记强度（FLI）至少比相应正常样本高2.7倍，可用于区分癌组织与正常及轻度发育异常组织（p<0.01）。综合分析，N/C比例和平均FLI可能会提高区分癌组织与正常鳞状上皮的能力。
