4 July 2000 Applications of two-photon fluorescence microscopy in deep-tissue imaging
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Based on the non-linear excitation of fluorescence molecules, two-photon fluorescence microscopy has become a significant new tool for biological imaging. The point-like excitation characteristic of this technique enhances image quality by the virtual elimination of off-focal fluorescence. Furthermore, sample photodamage is greatly reduced because fluorescence excitation is limited to the focal region. For deep tissue imaging, two-photon microscopy has the additional benefit in the greatly improved imaging depth penetration. Since the near- infrared laser sources used in two-photon microscopy scatter less than their UV/glue-green counterparts, in-depth imaging of highly scattering specimen can be greatly improved. In this work, we will present data characterizing both the imaging characteristics (point-spread-functions) and tissue samples (skin) images using this novel technology. In particular, we will demonstrate how blind deconvolution can be used further improve two-photon image quality and how this technique can be used to study mechanisms of chemically-enhanced, transdermal drug delivery.
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Chen-Yuan Dong, Chen-Yuan Dong, Betty Yu, Betty Yu, Lily Laiho Hsu, Lily Laiho Hsu, Peter D. Kaplan, Peter D. Kaplan, D. Blankschstein, D. Blankschstein, Robert Langer, Robert Langer, Peter T. C. So, Peter T. C. So, } "Applications of two-photon fluorescence microscopy in deep-tissue imaging", Proc. SPIE 4082, Optical Sensing, Imaging, and Manipulation for Biological and Biomedical Applications, (4 July 2000); doi: 10.1117/12.390532; https://doi.org/10.1117/12.390532

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