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Perfectly registered multiphoton and reflectance confocal video rate imaging of in vivo human skin
Author(s) -
Wang Hequn,
Lee Anthony M.D.,
Frehlick Zack,
Lui Harvey,
McLean David I.,
Tang Shuo,
Zeng Haishan
Publication year - 2013
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.201200067
Subject(s) - confocal , confocal microscopy , materials science , biomedical engineering , fluorescence lifetime imaging microscopy , human skin , optics , two photon excitation microscopy , in vivo , preclinical imaging , femtosecond , microscopy , laser , fluorescence , physics , medicine , biology , microbiology and biotechnology , genetics
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)