Iwasaki Shin-Ichi, Aoyagi Hidekazu
Advanced Research Center, The Nippon Dental University School of Life Dentistry at Niigata, Niigata, Japan.
Odontology. 2007 Jul;95(1):61-5. doi: 10.1007/s10266-007-0072-x. Epub 2007 Jul 25.
We examined the expression of keratin 14 (K14) on the lingual epithelium by immunofluorescent staining while monitoring morphological changes in the filiform papillae of mice by confocal laser-scanning microscopy in the transmission mode of the same sections to define both the histology and the morphology of cells. It is difficult to visualize histological details of the fetal lingual epithelium of the mouse on semi-ultrathin sections by light microscopy after immunohistochemical staining because the histological structures in such sections cannot be distinguished by standard counterstaining. To solve this problem and to visualize the immunoreactivity specific for K14, we analyzed the results of immunofluorescent staining of semi-ultrathin sections in combination with an examination of the corresponding images by laser-scanning microscopy in the transmission mode after staining of specimens with toluidine blue. No immunoreactivity specific for K14 was detected on the lingual epithelium of fetuses on embryonic day 15 (E15), but immunoreactivity was distinct at all postnatal stages from postnatal day 0 (P0) to P21.
我们通过免疫荧光染色检查了角蛋白14（K14）在舌上皮中的表达，同时在同一切片的透射模式下，利用共聚焦激光扫描显微镜监测小鼠丝状乳头的形态变化，以确定细胞的组织学和形态学特征。在免疫组织化学染色后，通过光学显微镜观察小鼠胎儿舌上皮的半超薄切片时，很难看到其组织学细节，因为在这样的切片中，组织学结构无法通过标准复染来区分。为了解决这个问题并可视化K14特异性免疫反应性，我们分析了半超薄切片的免疫荧光染色结果，并结合在标本用甲苯胺蓝染色后，通过透射模式的激光扫描显微镜检查相应图像。在胚胎第15天（E15）的胎儿舌上皮上未检测到K14特异性免疫反应性，但在出生后第0天（P0）至P21的所有出生后阶段，免疫反应性都很明显。
