Multiphoton microscopy imaging of developing tooth germs

Pei-Yu Pan; Rung-Shu Chen; Chih-Liang Ting; Wei-Liang Chen; Chen-Yuan Dong; Min-Huey Chen

doi:10.1016/j.jfma.2012.03.016

Multiphoton microscopy imaging of developing tooth germs

J Formos Med Assoc. 2014 Jan;113(1):42-9. doi: 10.1016/j.jfma.2012.03.016. Epub 2012 Jun 5.

Authors

Pei-Yu Pan¹, Rung-Shu Chen², Chih-Liang Ting¹, Wei-Liang Chen³, Chen-Yuan Dong³, Min-Huey Chen⁴

Affiliations

¹ School of Dentistry, National Taiwan University, Taipei, Taiwan, ROC.
² Graduate Institute of Clinical Dental Science, School of Dentistry, National Taiwan University, Taipei, Taiwan, ROC.
³ Department of Physics and Institute of Biotechnology, National Taiwan University, Taipei, Taiwan, ROC.
⁴ School of Dentistry, National Taiwan University, Taipei, Taiwan, ROC; Graduate Institute of Clinical Dental Science, School of Dentistry, National Taiwan University, Taipei, Taiwan, ROC; Department of Dentistry, National Taiwan University Hospital, Taipei, Taiwan, ROC. Electronic address: minhueychen@ntu.edu.tw.

PMID: 24445010
DOI: 10.1016/j.jfma.2012.03.016

Abstract

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.

Methods: Tooth germs were isolated from embryonic and newborn mice with age of Embryonic Day 14.5 and Postnatal Days 1, 3, 5, and 7.

Results: 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.

Conclusion: 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.

Keywords: autofluorescence; dentistry; multiphoton laser fluorescence microscope; second-harmonic generation signal; tooth germ.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Animals, Newborn
Mice
Microscopy, Fluorescence, Multiphoton*
Tooth Germ / cytology*
Tooth Germ / embryology