Background: High-resolution images of the epidermis are important to understand the transdermal penetration and changes in epidermal components. Both ex vivo and in vivo technologies are available to picture the epidermal thickness (ET). So far, the illustration of the stratum corneum (SC) has not been possible without artifacts. Objective: Precision in vivo measurement of the ET and SC, duly considering the impact of location on the body, age, and gender. Methods: In this pilot study on 20 skin-healthy subjects aged 18–66 years, the ET was imaged by two-photon microscopy (2PM) and optical coherence tomography (OCT), and the SC by using 2PM at five different body sites. Results: On solar-exposed body areas, both the epidermis and SC are thicker compared to solar-protected areas (p < 0.05), the epidermis at the gluteal region being the thickest (p < 0.05). The ET decreases with age (p < 0.05). Males show a thicker epidermis than females (p < 0.05). Conclusion: 2PM provides a noninvasive method for imaging the epidermis and especially the SC in vivo and is optimally suited for the application of histological criteria.

Background/Aims: Extrinsic and intrinsic skin aging is subject to constant remodeling and degradation processes, primarily in components of the extracellular matrix. While collagen fibers thin out during the aging process, the amorphous elastin fibers accumulate. These are essential formative components of the dermis. So far, these processes have been detected in vertical histological sections of invasive biopsies and recently in noninvasive horizontal scans. Methods: In this pilot study, a modified noninvasive 2-photon microscope was applied to measure the collagen/elastin index of skin in vivo. The obtained images permit an immediate vertical survey and allow a conclusion on the dermal composition at once. The collagen/elastin index was quantified by the second harmonic to autofluorescence aging index of dermis (SAAID) depending on volunteers' age (18-66 years), gender, and body area. Results: The highest SAAID was measured at the volar forearm as compared to the abdominal SAAID, which was significantly lower (p < 0.05). The gluteal region showed the significantly lowest SAAID (p < 0.05). The SAAID in female skin was higher compared to male skin and decreased with increasing age. Conclusion: These effects are to be considered in subsequent studies to be able to specifically detect and evaluate influences.

The optical biopsy could be a quick and painless support or alternative to a punch biopsy. In this letter the first in vivo vertical wide field two photon microscopy (2PM) images of healthy volunteers are shown. The 2PM images are fused images of two photon excited auto fluorescence (AF) and second harmonic generation (SHG) signals given as false-color images of 200 μm  ×  7 mm in size. By using these two nonlinear effects, the epidermis can be easily distinguished from the dermis at a glance. The auto fluorescence provides cellular resolution of the epidermal cells, and elastin fibers are partly visible in the dermis. Collagen, visible by SHG signal, is the dominant structure in the dermis. As contact agent water was evaluated to increase the AF signal, especially in the deeper layers of epidermis and dermis. For further improvement any terminal hairs should be removed by shaving and by taking tape strips of the first five layers of the stratum corneum. The first images illustrated that young skin compared to aged skin shows remarkably different dermal elastin and collagen signals in the dermis.