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, Betty Yu, Lily Laiho Hsu, Peter D. Kaplan, D. Blankschstein, Robert Langer, 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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