Virtual histology of colorectal lesions using laser-scanning confocal microscopy.
Virtual histology of colorectal lesions using laser-scanning confocal microscopy.
复制标题
使用激光扫描共聚焦显微镜进行结直肠病变的虚拟组织学。
作者：
M. Sakashita;H. Inoue;H. Kashida;Jun‐ichi Tanaka;J. Cho;H. Satodate;E. Hidaka;Tatsuya Yoshida;N. Fukami;Y. Tamegai;A. Shiokawa;S. Kudo
背景和研究目的
胃肠道病变的组织学检查目前基于薄切片标本的光学显微镜检查，并用苏木精和伊红染色。进行了一项研究，使用激光扫描共焦显微镜 (LCM) 获取未经处理的标本的即时显微图像，以检查结直肠病变。还开发出了可穿过内窥镜工作通道的探头式LCM原型内窥镜。
材料和方法
研究材料包括在昭和大学北横滨医院通过内窥镜或手术切除的结直肠病变。使用 LCM 检查一百个未经处理的样本。病变组织病理学结果为：正常结肠粘膜7例，增生性息肉5例，低度不典型增生腺瘤68例，高度不典型增生腺瘤10例，腺癌10例。 LCM 研究使用波长为 488 nm 的氩激光束。使用内窥镜可以观察切除的正常结肠粘膜（体外）和健康志愿者的直肠粘膜（体内）。将每个样本的 LCM 图像与苏木精-伊红染色的组织病理学横截面进行比较。
结果
LCM 图像与传统的苏木精-伊红光显微图像吻合良好。在正常粘膜或增生性息肉中看不到细胞核。在具有高度不典型增生和癌的腺瘤中，细胞核比具有低度不典型增生的腺瘤更常见。两组之间的细胞核可视化率存在显着差异（P < 0.01）（60.0 % vs. 10.3 %）。在使用内窥镜的 LCM 图像中，可以在体外和体内识别结肠腺和杯状细胞的孔口。
结论
激光扫描共焦显微镜可提供与苏木精-伊红染色的图像良好对应的即时图像。一种改进的探针型 LCM 内窥镜正在开发中，它可以提供更好的体内结直肠病变组织学图像。
BACKGROUND AND STUDY AIMS
Histological examination of gastrointestinal lesions is currently based on light-microscopic examination of thin-slice specimens, with hematoxylin and eosin staining. A study of the use of laser-scanning confocal microscopy (LCM) to obtain immediate microscopic images of untreated specimens for examining colorectal lesions was carried out. A probe-type LCM prototype endomicroscope that can be passed through the working channel of an endoscope has also been developed.
MATERIALS AND METHODS
The study materials consisted of colorectal lesions resected either endoscopically or surgically at Showa University Northern Yokohama Hospital. One hundred untreated specimens were examined using LCM. The histopathological findings in the lesions were seven cases of normal colonic mucosa, five hyperplastic polyps, 68 adenomas with low-grade dysplasia, 10 adenomas with high-grade dysplasia, and 10 adenocarcinomas. An argon laser beam with a wavelength of 488 nm was used for the LCM study. Observation of the resected normal colonic mucosa (in vitro) and the rectal mucosa of a healthy volunteer (in vivo) was possible using the endomicroscope. The LCM images for each specimen were compared with the hematoxylin-eosin-stained histopathological cross-sections.
RESULTS
The LCM images corresponded well with the conventional hematoxylin-eosin light-microscopic images. The nuclei were not visualized in normal mucosa or hyperplastic polyps. In adenomas with high-grade dysplasia and carcinomas, nuclei were more often visible than in adenomas with low-grade dysplasia. The rate of visualization of nuclei was significantly different ( P < 0.01) between these two groups (60.0 % vs. 10.3 %). In LCM images using endomicroscope, it was possible to recognize the orifices of the colonic glands and goblet cells both in vitro and in vivo.
CONCLUSIONS
Laser-scanning confocal microscopy provides immediate images that correspond well with those of hematoxylin-eosin staining. An improved probe-type LCM endomicroscope is being developed which should provide better histological images of colorectal lesions in vivo.