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Multifunctional Imaging of Endogenous Contrast by Simultaneous Nonlinear and Optical Coherence Microscopy of Thick Tissues
Author(s) -
Yazdanfar Siavash,
Chen Yen Yu,
So Peter T.C.,
Laiho Lily H.
Publication year - 2007
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.20447
Subject(s) - microscopy , autofluorescence , two photon excitation microscopy , second harmonic generation , materials science , confocal microscopy , microscope , optics , optical coherence tomography , elastin , fluorescence microscope , fluorescence , laser , pathology , physics , medicine
A variety of high resolution optical microscopy techniques have been developed in recent years for basic and clinical studies of biological systems. We demonstrate a trimodal microscope combining optical coherence microscopy (OCM) with two forms of nonlinear microscopy, namely two‐photon excited fluorescence (2PF) and second harmonic generation (SHG), for imaging turbid media. OCM combines the advantages of confocal detection and coherence gating for structural imaging in highly scattering tissues. Nonlinear microscopy enables the detection of biochemical species, such as elastin, NAD(P)H, and collagen. While 2PF arises from nonlinear excitation of fluorescent species, SHG is a form of nonlinear scattering observed in materials that lack a center of inversion symmetry, such as type I collagen. Characterization of the microscope showed nearly diffraction‐limited spatial resolution in all modalities. Images were obtained in fish scales and excised human skin samples. The primary endogenous sources of contrast in the dermis were due to elastin autofluorescence and collagen SHG. Multimodal microscopy allows the simultaneous visualization of structural and functional information of biological systems. Microsc. Res. Tech., 2007. © 2007 Wiley‐Liss, Inc.