Abstract
Confocal and nonlinear optical microscopies have been applied for dermatological studies because of their capability to provide sub-surface three-dimensional images with sub-microm spatial resolutions. Optical signal degradation as the imaging plane being moved toward deeper regions in skin specimens is the key factor that limits the observation depth for the laser scanning based linear or nonlinear imaging modalities. In this article, we studied the signal degradation in fixed human skin specimens using reflection confocal microscopy and higher-harmonic optical microscopy based on a Cr:forsterite femtosecond laser centered at 1230-nm. By analyzing the optical properties through these linear and nonlinear imaging modalities, we found that the optical signal degradation in the studied human skin specimen is dominated by the distortion of the point spread function.
Highlights
Optical imaging and microscopy such as optical coherence tomography (OCT) [1,2,3], confocal microscopy [4,5], and nonlinear optical microscopy [6-10] have been widely applied for dermatological studies because of their high penetration and three-dimensional (3D) imaging capability with high spatial resolutions
We studied the signal degradation in fixed human skin specimens using reflection confocal microscopy and higher-harmonic optical microscopy based on a Cr:forsterite femtosecond laser centered at 1230-nm
By analyzing the optical properties through these linear and nonlinear imaging modalities, we found that the optical signal degradation in the studied human skin specimen is dominated by the distortion of the point spread function
Summary
Optical imaging and microscopy such as optical coherence tomography (OCT) [1,2,3], confocal microscopy [4,5], and nonlinear optical microscopy [6-10] have been widely applied for dermatological studies because of their high penetration and three-dimensional (3D) imaging capability with high spatial resolutions. Compared with OCT, laser scanning confocal and nonlinear microscopies provide much higher lateral resolution while the sectioned two-dimensional images are in the plane parallel to the surface, in contrast to the cross-sectional (tangential) images provided by OCT. Two-photon fluorescence microscopy [14] of skin based on 780-nm femtosecond light provides high resolution imaging from the skin surface through the epidermal-dermal junction [7]. We demonstrated that higher-harmonic optical microscopy (HOM) based on a 1230-nm femtosecond light source could provide sub-micrometer-resolution deep-tissue biopsy images of skin without the use of fluorescence or exogenous markers [9,10], which could be considered as a powerful tool for dermatological studies
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