Pan Pei-Yu, Chen Rung-Shu, Ting Chih-Liang, Chen Wei-Liang, Dong Chen-Yuan, Chen Min-Huey
School of Dentistry, National Taiwan University, Taipei, Taiwan, ROC.
Graduate Institute of Clinical Dental Science, School of Dentistry, National Taiwan University, Taipei, Taiwan, ROC.
J Formos Med Assoc. 2014 Jan;113(1):42-9. doi: 10.1016/j.jfma.2012.03.016. Epub 2012 Jun 5.
BACKGROUND/PURPOSE: Traditionally, tooth germ is observed by histological investigation with hematoxylin and eosin stain and information may loss during the process. The purpose of this study is to use multiphoton laser fluorescence microscopy to observe the developing tooth germs of mice for building up the database of the images of tooth germs and compare with those from conventional histological analysis.
Tooth germs were isolated from embryonic and newborn mice with age of Embryonic Day 14.5 and Postnatal Days 1, 3, 5, and 7.
Comparison of the images of tooth germ sections in multiphoton microscopy with the images of histology was performed for investigating the molar tooth germs. It was found that various signals arose from different structures of tooth germs. Pre-dentin and dentin have strong second-harmonic generation signals, while ameloblasts and enamel tissues were shown with strong autofluorescence signals.
In this study, a novel multiphoton microscopy database of images from developing tooth germs in mice was set up. We confirmed that multiphoton laser microscopy is a powerful tool for investigating the development of tooth germ and is worthy for further application in the study of tooth regeneration.
背景/目的：传统上，牙胚是通过苏木精和伊红染色的组织学研究来观察的，在此过程中信息可能会丢失。本研究的目的是使用多光子激光荧光显微镜观察小鼠正在发育的牙胚，以建立牙胚图像数据库，并与传统组织学分析的图像进行比较。
从胚胎期第14.5天以及出生后第1、3、5和7天的胚胎和新生小鼠中分离出牙胚。
对多光子显微镜下牙胚切片的图像与组织学图像进行比较，以研究磨牙牙胚。发现牙胚的不同结构产生了各种信号。前期牙本质和牙本质具有强烈的二次谐波产生信号，而成釉细胞和釉质组织则显示出强烈的自发荧光信号。
在本研究中，建立了一个来自小鼠正在发育的牙胚的新型多光子显微镜图像数据库。我们证实多光子激光显微镜是研究牙胚发育的有力工具，值得在牙齿再生研究中进一步应用。
