Two-photon 3D mapping of tissue endogenous fluorescence species based on fluoresence excitation spectra

Lily Laiho Hsu; Thomas M. Hancewicz; Peter D. Kaplan; Keith M. Berland; Peter T. C. So

doi:10.1117/12.424567

24 April 2001 Two-photon 3D mapping of tissue endogenous fluorescence species based on fluoresence excitation spectra

Lily Laiho Hsu, Thomas M. Hancewicz, Peter D. Kaplan, Keith M. Berland, Peter T. C. So

Proceedings Volume 4262, Multiphoton Microscopy in the Biomedical Sciences; (2001) https://doi.org/10.1117/12.424567
Event: BiOS 2001 The International Symposium on Biomedical Optics, 2001, San Jose, CA, United States

Abstract

Deep tissue imaging may have important biomedical applications in the areas of skin disease diagnosis, wound healing, and tissue engineering. For the study oftissue physiology with microscopic resolution, we used two-photon microscopic imaging based on the excitation of endogenous fluorophores. While autofluorescence is observed ubiquitously in many tissue types, the identities and distributions of these fluorophores have not been completely characterized. The different fluorescent species are expected to have different fluorescence excitation and emission spectra. Self-modeling curve resolution (SMCR) can be applied to extract spectroscopic components from two-photon images. In ex vivo human skin, we were able to acquire a four-dimensional data set (3D space + excitation spectra). We extracted the major spectral components from this data set using multivariate curve resolution and correlated these species with known tissue structures. From the SMCR analysis, it was determined that there are approximately seven factors that contribute to most of the autofluorescence from human skin. This analysis provides us with the concentration ofthe species at different depths within the skin and also with a reconstructed image of the skin due to each single factor alone. Several ofthese chemical components have been identified, such as collagen, elastin, and NAD(P)H. In addition to providing insight into tissue physiology, we are able to optimize the excitation wavelength for each biochemical species for skin imaging applications.

Citation Download Citation

Lily Laiho Hsu, Thomas M. Hancewicz, Peter D. Kaplan, Keith M. Berland, and Peter T. C. So "Two-photon 3D mapping of tissue endogenous fluorescence species based on fluoresence excitation spectra", Proc. SPIE 4262, Multiphoton Microscopy in the Biomedical Sciences, (24 April 2001); https://doi.org/10.1117/12.424567

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