Confocal scanning laser microscopy (CSLM) represents a novel imaging technique for in vivo microscopic analysis of skin lesions at a level of resolution that allows morphologic analysis of microanatomic structures. We investigated the feasibility of recognizing the cellular constituents of pigmented skin lesions, such as pigmented keratinocytes, melanocytes, and melanophages, by CSLM. Fifteen pigmented lesions (five pigmented seborrheic keratoses, and 10 compound melanocytic nevi) from 15 patients were studied, as well as normal skin. After the clinical lesions were imaged by CSLM, they were biopsied or excised for examination by conventional histology for comparison of the morphologic features. In images obtained by CSLM, pigmented keratinocytes were seen as polygonal cohesive cells with variably bright granular cytoplasm. Melanocytes appeared as bright round, oval, fusiform, or dendritic cells. The architectural growth pattern of melanocytes could be analyzed. Melanocytes were identified by their nested growth pattern as aggregates of bright round to oval structures at the dermoepidermal junction or in the superficial dermis. Melanocytes were also recognizable as single cells along the dermoepidermal junction, usually separated from each other by a variable number of keratinocytes. Melanophages appeared as large bright plump cells with ill-defined cytoplasmic borders, usually located around or near vessels of the superficial dermis. Our results demonstrate that the cellular constituents of pigmented lesions can be recognized by CSLM. This technique sets a new paradigm for noninvasive quasihistologic examination of pigmented lesions in vivo and merits further evaluation for diagnostic use.

Ex vivo confocal scanning laser microscopy can potentially expedite Mohs surgery in rapidly detecting large nodular basal cell carcinomas without conventional frozen histopathology. Further improvements in instrumentation and image quality are necessary to allow broader application and acceptance of this novel technology in Mohs surgery.

The early detection of malignant melanoma remains challenging for physicians. New techniques are being explored in order to improve diagnostic accuracy. Confocal scanning laser microscopy (CSLM) represents one such novel imaging modality. It allows in vivo microscopic analysis of skin lesions at a level of resolution approaching histological detail. Therefore, interpretation of optical sections represents in principle a histopathological analysis. Pigmented lesions are particularly amenable to examination by CSLM, since melanin pigment provides endogenous contrast, facilitating the recognition of melanocytes and their distribution within the epidermis. As a first step to explore the use of CSLM in the detection of melanoma, we sought to determine whether images obtained by CSLM are suitable for analysis by established histopathological criteria for the diagnosis of melanoma. We examined five pigmented lesions clinically suspicious for melanoma from five individual patients. Following imaging by CSLM, the clinical lesions were excised for examination by conventional histology. The melanocytes in the confocal images were recognized within the epidermis by their bright cytoplasmic signal intensity. They were round to oval in shape and frequently showed dendritic processes of various lengths. Confocal images of melanoma showed an increased number of intraepidermal melanocytes in solitary units at all layers of the epidermis, including the upper spinous and granular cell layers. Our results demonstrate that intraepidermal melanoma can be recognized by CSLM through analysis of the intraepidermal growth patterns of melanocytes using the same criteria as established for conventional histology. Thus, the application of CSLM represents a new tool for non-invasive screening of intraepidermal pigmented lesions in vivo and offers the opportunity to bring histopathological analysis to the bedside.

Background Venous leg ulcers are responsible for more than half of all lower extremity ulcerations. Significant interest has been focused on understanding the physiologic basis upon which patients fail to heal with standard therapy. Objective This study uses complementary DNA microarray analysis of tissue samples from healing and non-healing venous leg ulcers to identify the genetic expression profiles from these dichotomous populations. Methods Ulcer size and chronicity, factors that have been identified as prognostic indicators for healing, were used to distribute venous leg ulcers as healing versus non-healing. Punch biopsy samples were obtained from the wound edge and wound bed of all venous leg ulcers. The top fifteen genes with differential expression greater than twofold between the two populations of wounds (p < 0.05) were reported. Results Significant differences were demonstrated in the expression of a diverse collection of genes, with particular differences demonstrated by genes coding for structural epidermal proteins, genes associated with hyperproliferation and tissue injury, as well as transcription factors. Limitations Small sample size may mitigate potential clinical implications of findings. Conclusions The genetic expression profiles displayed here may have implications for the development of novel therapies for chronic venous leg ulcers, and may also serve as prognostic indicators for wound healing.

Etanercept is an alternative treatment option for patients with refractory ulcers due to PG.