Skin optics revisited by in vivo confocal microscopy: melanin and sun exposure

A new confocal prototype dedicated to the exploration of in vivo human skin has been constructed around a laser confocal module (Oz Noran, Inc.) and a skin contact device, assuring perfect stability of skin images. The power of the Argon/Krypton laser source has been limited to 2mW to secure safety, and the laser provides three visible wavelengths: 488, 568, and 647 nm. Optical sections were digitized at video rate, providing easy and rapid measurements of the thickness of epidermal layers and time-resolved information. Unexpected details of the epidermis were recorded with the blue laser line. Melanin provided strong reflection of the basal keratinocytes instead of the absorption expected. The 3D reconstruction of the melanin cap in basal keratinocytes confirmed the behavior of melanosomes acting as myriads of nanomirrors that reflected light. Confocal images of the posterior aspect of the forearm were recorded before sun exposure and then for one month after exposure. There was a 25% increase in the thickness of the stratum corneum. Bright inclusions into the dark nucleus of numerous spinous cells were interpreted as local condensation of chromatin. Numerous bright intercellular filaments were attributed to melanosomes filling up dendrites of melanocytes. A striking observation concerned the lack of melanosome caps in basal keratinocytes. In vivo confocal microscopy affords new insight to the role of melanin and its gradual migration after sun exposure.