Three-dimensional skin imaging using the combination of reflected confocal and multiphoton microscopy

Reflected confocal microscopy has been widely used in clinical application in dermatology. In recent years, multiphoton microscopy has also emerged as an important minimally invasive bioimaging technique for the skin. In this study, we combine reflected confocal microscopy and multiphoton microscopy for skin imaging. In the epidermis, reflected confocal signals are expected to help in delineating cell borders while multiphoton signals provide cytoplasmic morphologies. In the dermis, second harmonic generation signals provide the morphology of collagen fibers. When three-dimensional images are projected, the detailed distribution of cellular component and extracellular matrix in skin can be obtained. Properly developed, this technique is of great potential for in vivo clinical application.