Multiphoton microscopy imaging of developing tooth germs.
OBJECTIVE
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.


CONCLUSIONS
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.
School of Dentistry, National Taiwan University, Taipei, Taiwan, ROC.
Pei-Yu
PY
Pan
Graduate Institute of Clinical Dental Science, School of Dentistry, National Taiwan University, Taipei, Taiwan, ROC.
Rung-Shu
RS
Chen
School of Dentistry, National Taiwan University, Taipei, Taiwan, ROC.
Chih-Liang
CL
Ting
Department of Physics and Institute of Biotechnology, National Taiwan University, Taipei, Taiwan, ROC.
Wei-Liang
WL
Chen
Department of Physics and Institute of Biotechnology, National Taiwan University, Taipei, Taiwan, ROC.
Chen-Yuan
CY
Dong
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.
Min-Huey
MH
Chen
D000818:Animals; D000831:Animals, Newborn; D051379:Mice; D036641:Microscopy, Fluorescence, Multiphoton; D014083:Tooth Germ / Q000166:cytology* / Q000196:embryology
autofluorescence; dentistry; multiphoton laser fluorescence microscope; second-harmonic generation signal; tooth germ