An objective measurement of cutaneous topographical information is important for quantifying the degree of skin aging. Our aim was to improve methods for measuring microrelief patterns using a three-dimensional analysis based on silicone replicas and scanning electron microscope (SEM). Another objective was to compare the results with those obtained using a two-dimensional analysis method based on dermoscopy. Silicone replicas were obtained from forearms, dorsum of the hands and fingers of 51 volunteers. Cutaneous profiles obtained by SEM with silicone replicas showed more consistent correlations with age than data obtained by dermoscopy. This indicates the advantage of three-dimensional topography analysis using silicone replicas and SEM over the widely used dermoscopic assessment. The cutaneous age was calculated using stepwise linear regression, and the result was 57.40-9.47 × (number of furrows on dorsum of the hand) × (width of furrows on dorsum of the hand).

The thickness of specimen is an important factor in microscopic researches. Thicker specimen contains more information, but it is diffi cult to obtain well focused image with precise details due to optical limit of conventional microscope. Recently, a microscope unit that combines improved illumination system, which allows real time three-dimensional (3D) image and automatic z-stack merging software. In this research, we evaluated the usefulness of this unit in observing thick samples; Golgi stained nervous tissue and ground prepared bone, tooth, and non-transparent small sample; zebra fish teeth. Well focused image in thick samples was obtained by processing z-stack images with Panfocal software. A clear feature of neuronal dendrite branching pattern could be taken. 3D features were clearly observed by oblique illumination. Furthermore, 3D array and shape of zebra fi sh teeth was clearly distinguished. A novel combination of two channel oblique illumination and z-stack imaging process increased depth of fi eld and optimized contrast, which has a potential to be further applied in the fi eld of neuroscience, hard tissue biology, and analysis of small organic structures such as ear ossicles and zebra fi sh teeth.