KEYWORDS: Skin, Two photon excitation microscopy, In vivo imaging, Collagen, Confocal laser scanning microscopy, 3D image processing, Microscopy, Confocal microscopy, Image analysis, 3D acquisition
We present results from 2D Fourier analysis on 3D stacks of images obtained by confocal laser scanning reflectance microscopy (CLSM) and two-photon fluorescence microscopy (2PM) on human skin in vivo. CLSM images were obtained with a modified commercial system (Vivascope1000, Lucid Inc, excitation wavelength 830 nm) equipped with a piezo-focusing element (350 μm range) for depth positioning of the objective lens. 2PM was performed with a specially designed set-up with excitation wavelength 730 nm. Mean cell size in the epidermal layer and structural orientation in the dermal layer have been determined as a function of depth by 2D Fourier analysis. Fourier analysis on microscopic images enables automatic non-invasive quantitative structural analysis (mean cell size and orientation) of living human skin.
Optical coherence tomography (OCT) and confocal laser scanning microscopy (CLSM) were applied to characterize non-invasively and in vivo the upper layers of human skin on the back of the
hand. The techniques enable a detailed determination of the thickness and location of various skin layers in the epidermis and superficial dermis. Due to differences in spatial resolution and
penetration depth of these methods, OCT and CLSM give complementary information on the composition and structure of skin. OCT signals of the back of the hand show three reflecting layers at different depth in the skin. A direct comparison with CLSM enables the assignment of these layers: the first one is due to the reflection at the skin surface, the second one appears to be caused by the reflection at the basal epidermal layer and the third layer can be ascribed to reflection at fibrous structure in the upper dermis. A comparison of methods reveals a consistent interpretation of the images.
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