Al-Arashi Munir Y, Salomatina Elena, Yaroslavsky Anna N
Harvard Medical School, Boston, Massachusetts 02114, USA.
Lasers Surg Med. 2007 Oct;39(9):696-705. doi: 10.1002/lsm.20578.
The standard diagnostic procedure for skin cancers is invasive biopsy followed by histopathological evaluation. The biopsy may result in scarring and infection. A reliable way to noninvasively image suspicious lesions with high resolution and contrast would be valuable. In this study, the suitability of dye-enhanced multimodal confocal microscopy for the detection of nonmelanoma skin cancers was evaluated.
For the experiments we used fresh tumor material stained using 0.2 mg/ml or 0.05 mg/ml aqueous solutions of methylene blue (MB) or toluidine blue (TB), respectively. Reflectance, fluorescence, and fluorescence polarization images of skin specimens stained with MB and TB were excited by 656 nm and 633 nm light, respectively. Fluorescence emission and anisotropy were registered between 690 nm and 710 nm. In addition, reference reflectance images at 830 nm were acquired. In total we imaged, analyzed, and compared to histology at least 10 samples of each tumor-type including nodular basal cell carcinoma (BCC), infiltrative basal cell carcinoma, and squamous cell carcinoma (SCC).
The morphological features and appearance of skin structures in the fluorescence images correlate well with corresponding histology for all investigated tumor-types. Multi-spectral reflectance images provide information on the tissue spectral responses and are complimentary to the fluorescence images. The differences detected by fluorescence polarization in cancerous and normal structures may be used for cancerous tissue discrimination. Our results indicate the feasibility of using multimodal confocal microscopy as real-time tool for detecting skin pathology.
皮肤癌的标准诊断程序是进行侵入性活检，随后进行组织病理学评估。活检可能会导致瘢痕形成和感染。一种能够以高分辨率和对比度对可疑病变进行无创成像的可靠方法将具有重要价值。在本研究中，评估了染料增强多模态共聚焦显微镜用于检测非黑素瘤皮肤癌的适用性。
在实验中，我们分别使用0.2毫克/毫升或0.05毫克/毫升的亚甲蓝（MB）或甲苯胺蓝（TB）水溶液对新鲜肿瘤材料进行染色。用656纳米和633纳米的光分别激发用MB和TB染色的皮肤标本的反射率、荧光和荧光偏振图像。在690纳米至710纳米之间记录荧光发射和各向异性。此外，还采集了830纳米处的参考反射率图像。我们总共对每种肿瘤类型（包括结节性基底细胞癌（BCC）、浸润性基底细胞癌和鳞状细胞癌（SCC））的至少10个样本进行了成像、分析，并与组织学进行了比较。
对于所有研究的肿瘤类型，荧光图像中皮肤结构的形态特征和外观与相应的组织学结果具有良好的相关性。多光谱反射率图像提供了有关组织光谱响应的信息，并且是对荧光图像的补充。通过荧光偏振在癌组织和正常组织中检测到的差异可用于癌组织的鉴别。我们的结果表明，使用多模态共聚焦显微镜作为检测皮肤病理的实时工具是可行的。
