Compound imaging of ocular tissues with backward second harmonic generation and two-photon excitation fluorescence

Danni Chen; Junle Qu; Gaixia Xu; Lixin Liu; Ziyang Lin; Hanben Niu

doi:10.1117/12.710887

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27 October 2006 Compound imaging of ocular tissues with backward second harmonic generation and two-photon excitation fluorescence

Danni Chen, Junle Qu, Gaixia Xu, Lixin Liu, Ziyang Lin, Hanben Niu

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Proceedings Volume 6047, Fourth International Conference on Photonics and Imaging in Biology and Medicine; 604718 (2006) https://doi.org/10.1117/12.710887
Event: Fourth International Conference on Photonics and Imaging in Biology and Medicine, 2005, Tianjin, China

Abstract

We report the implementation of compound imaging with backward second harnionic generation (SHG) and two-photon excitation fluorescence (TPEF) in a standard confocal microscope. Various biological tissues, including skin, muscle and ocular tissues (e.g. cornea. iris, choroid, and sclera) have been investigated with different sample preparation methods (fresh, fixation). The results show that collagen-rich ocular tissues, like cornea, iris, choroids, and sclera can produce strong SHG signals. In sclera, there are not only plenty of collagen, but also various endogenous fluorophores. The organization of collagen fibers in sclera is clearly distinguishable from its SHG images viewed transversely and horizontally respectively. We also find that fixation of the sample with formaldehyde has reduced the intensity of SHG intensity by almost 5O%. As the SHG and TPEF can provide complementary information about collagen and fluorophores rich biological tissues, compound imaging of SHG and TPEF presented in this study is believed to have potential applications in biomedicine and clinical diagnosis.

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Danni Chen, Junle Qu, Gaixia Xu, Lixin Liu, Ziyang Lin, and Hanben Niu "Compound imaging of ocular tissues with backward second harmonic generation and two-photon excitation fluorescence", Proc. SPIE 6047, Fourth International Conference on Photonics and Imaging in Biology and Medicine, 604718 (27 October 2006); https://doi.org/10.1117/12.710887

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