Virtual biopsy of rat tympanic membrane using higher harmonic generation microscopy.
Multiharmonic optical microscopy has been widely applied in biomedical research due to its unique capability to perform noninvasive studies of biomaterials. In this study, virtual biopsy based on back-propagating multiple optical harmonics, combining second and third harmonics, is applied in unfixed rat tympanic membrane. We show that third harmonic generation can provide morphologic information on the epithelial layers of rat tympanic membrane as well as radial collagen fibers in middle fibrous layers, and that second harmonic generation can provide information on both radial and circular collagen fibers in middle fibrous layers. Through third harmonic generation, the capillary and red blood cells in the middle fibrous layer are also noted. Additionally, the 3-D relationship to adjacent bony structures and spatial variations in thickness and curvature are obtained. Our study demonstrates the feasibility of using a noninvasive optical imaging system for comprehensive evaluation of the tympanic membrane.
Lee
Wen-Jeng
WJ
National Taiwan University, Department of Electrical Engineering, Taipei, Taiwan.
Lee
Chia-Fone
CF
Chen
Szu-Yu
SY
Chen
Yuh-Shyang
YS
Sun
Chi-Kuang
CK
D000465:Algorithms; D000818:Animals; D001706:Biopsy / Q000379:methods*; D007089:Image Enhancement / Q000379:methods; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D010363:Pattern Recognition, Automated / Q000379:methods*; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014432:Tympanic Membrane / Q000166:cytology*; D014584:User-Computer Interface