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Perfectly registered multiphoton and reflectance confocal video rate imaging of in vivo human skin
Authors:Zack Frehlick  Harvey Lui  David I. McLean  Shuo Tang  Haishan Zeng
Affiliation:1. Imaging Unit – Integrative Oncology Department, British Columbia Cancer Agency Research Centre, Vancouver, BC, Canada;2. Photomedicine Institute, Department of Dermatology and Skin Science, University of British Columbia and Vancouver Coastal Health Research Institute, Vancouver, BC, Canada;3. Department of Electrical and Computer Engineering, University of British Columbia, Vancouver, BC, Canada;4. Department of Physics and Astronomy, University of British Columbia, Vancouver, BC, Canada
Abstract:We present a multimodal in vivo skin imaging instrument that is capable of simultaneously acquiring multiphoton and reflectance confocal images at up to 27 frames per second with 256 × 256 pixel resolution without the use of exogenous contrast agents. A single femtosecond laser excitation source is used for all channels ensuring perfect image registration between the two‐photon fluorescence (TPF), second harmonic generation (SHG), and reflectance confocal microscopy (RCM) images. Images and videos acquired with the system show that the three imaging channels provide complementary information in in vivo human skin measurements. In the epidermis, cell boundaries are clearly seen in the RCM channel, while cytoplasm is better seen in the TPF imaging channel, whereas in the dermis, SHG and TPF channels show collagen bundles and elastin fibers, respectively. The demonstrated fast imaging speed and multimodal imaging capabilities of this MPM/RCM instrument are essential features for future clinical application of this technique. (© 2013 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Keywords:multiphoton microscopy  reflectance confocal microscopy  in vivo video microscopy  skin diagnosis
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