White W M, Rajadhyaksha M, González S, Fabian R L, Anderson R R
Otolaryngology/Head and Neck Surgery, Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, USA.
Laryngoscope. 1999 Oct;109(10):1709-17. doi: 10.1097/00005537-199910000-00029.
OBJECTIVES/HYPOTHESIS: To study the microscopic anatomy of normal oral tissues in vivo using confocal reflectance microscopy (CRM). This novel and noninvasive imaging modality can define and characterize healthy oral mucosa and thus this work serves as the foundation for studying oral diseases in vivo.
This was a pilot observational cohort study comparing noninvasive CRM images with histology.
Lip and tongue mucosa were imaged by CRM in six healthy human subjects. In CRM living tissue is illuminated by a laser source and backscattered (or reflected) light is collected by a detector. Image contrast is determined by natural differences in refractive indices of organelles and other subcellular structures within the tissues. Gray-scale images were displayed in real-time on a video monitor and represented horizontal (en face) optical sections through the tissue. Motion of the oral tissue relative to the objective lens was minimized with a tissue stabilizer. After imaging, biopsies were taken from the same site of lip mucosa to correlate noninvasive confocal images with conventional histology.
Confocal images correlated well with conventional histology, both qualitatively (visual analysis) and quantitatively (stereology). Imaging was possible up to depths of 490 and 250 microm in the lip and tongue, respectively. Cells and organelles including nuclei, circulating blood cells, and extracellular matrix were clearly observed.
CRM provides details of normal human oral mucosa at the cellular level without the artifacts of histological processing, and thus has the potential for further development and use in clinical practice as a diagnostic tool for the early detection of oral cancer and precancer.
目的/假设:使用共聚焦反射显微镜(CRM)研究体内正常口腔组织的微观解剖结构。这种新颖的非侵入性成像方式能够定义和表征健康的口腔黏膜,因此这项工作为体内口腔疾病的研究奠定了基础。
这是一项将非侵入性CRM图像与组织学进行比较的试点观察性队列研究。
对6名健康人类受试者的唇部和舌部黏膜进行CRM成像。在CRM中,激光源照射活体组织,探测器收集反向散射(或反射)光。图像对比度由组织内细胞器和其他亚细胞结构的折射率自然差异决定。灰度图像实时显示在视频监视器上,代表穿过组织的水平(正面)光学切片。使用组织稳定器将口腔组织相对于物镜的运动降至最低。成像后,从唇部黏膜的同一部位取活检组织,以将非侵入性共聚焦图像与传统组织学相关联。
共聚焦图像在定性(视觉分析)和定量(体视学)方面均与传统组织学良好相关。在唇部和舌部分别可达490和250微米深度进行成像。清晰观察到包括细胞核、循环血细胞和细胞外基质在内的细胞和细胞器。
CRM可在细胞水平提供正常人类口腔黏膜的详细信息,而无组织学处理的伪像,因此有进一步发展并作为口腔癌和癌前病变早期检测的诊断工具应用于临床实践的潜力。