Yeh Alvin T, Nassif Nader, Zoumi Aikaterini, Tromberg Bruce J
Opt Lett. 2002 Dec 2;27(23):2082-4. doi: 10.1364/ol.27.002082.
A multiphoton microscope employing second-harmonic generation (SHG) and two-photon excited fluorescence (TPF) is used for high-resolution ex vivo imaging of rabbit cornea in a backscattering geometry. Endogenous TPF and SHG signals from corneal cells and extracellular matrix, respectively, are clearly visible without exogenous dyes. Spectral characterization of these upconverted signals provides confirmation of the structural origin of both TPF and SHG, and spectral imaging facilitates the separation of keratocyte and epithelial cells from the collagen-rich corneal stroma. The polarization dependence of collagen SHG is used to highlight fiber orientation, and three-dimensional SHG tomography reveals that approximately 88% of the stromal volume is occupied by collagen lamellae.
一种采用二次谐波产生（SHG）和双光子激发荧光（TPF）的多光子显微镜，用于在背散射几何结构下对兔角膜进行高分辨率离体成像。分别来自角膜细胞和细胞外基质的内源性TPF和SHG信号在无需外源性染料的情况下清晰可见。这些上转换信号的光谱表征证实了TPF和SHG的结构起源，并且光谱成像有助于从富含胶原蛋白的角膜基质中分离角膜细胞和上皮细胞。利用胶原蛋白SHG的偏振依赖性来突出纤维方向，三维SHG断层扫描显示约88%的基质体积被胶原板层占据。
