Abstract
We demonstrate multimodal label-free nonlinear optical microscopy in human skin enabled by a fiber-based two-color ultrafast source. Energetic femtosecond pulses at 775 nm and 1250 nm are simultaneously generated by an Er-fiber laser source employing frequency doubling and self-phase modulation enabled spectral selection. The integrated nonlinear optical microscope driven by such a two-color femtosecond source enables the excitation of endogenous two-photon excitation fluorescence, second-harmonic generation, and third-harmonic generation in human skin. Such a 3-channel imaging platform constitutes a powerful tool for clinical application and optical virtual skin biopsy.
Highlights
Nonlinear optical microscopy (NLOM) is one of the most important label-free techniques to conduct optical virtual skin biopsy [1]
We demonstrate multimodal label-free nonlinear optical microscopy in human skin enabled by a fiber-based two-color ultrafast source
To demonstrate the capability of our multimodal platform for multiphoton label-free imaging in human skin, we conduct second-harmonic generation (SHG)/third-harmonic generation (THG) microscopy excited by 1250-nm pulses and 2PEF microscopy excited by 775-nm pulses
Summary
Nonlinear optical microscopy (NLOM) is one of the most important label-free techniques to conduct optical virtual skin biopsy [1]. Arising from interfaces and optical inhomogeneity, THG can reveal the cell outline and differentiate stratum corneum (SC), stratum granulosum (SG), stratum spinosum (SS), and stratum basale (SB) [13,15,16]. These modalities are powerful bioimaging tools for histopathology, morphology, and disease diagnosis. Besides imaging tumors [28,29], THG is able to measure the nuclear-cytoplasmic (NC) ratio—an important index for scoring skin aging [30]
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