Abstract

In vivo imaging of pigmented lesions in human skin was performed with a clinical multiphoton microscopy (MPM)-based tomograph (MPTflex, JenLab, Germany). Two-photon excited fluorescence was used for visualizing endogenous fluorophores such as NADH/FAD, keratin, melanin in the epidermal cells and elastin fibers in the dermis. Collagen fibers were imaged by second harmonic generation. Our study involved in vivo imaging of benign melanocytic nevi, atypical nevi and melanoma. The goal of this preliminary study was to identify in vivo the characteristic features and their frequency in pigmented lesions at different stages (benign, atypical and malignant) and to evaluate the ability of in vivo MPM to distinguish atypical nevi from melanoma. Comparison with histopathology was performed for the biopsied lesions. Benign melanocytic nevi were characterized by the presence of nevus cell nests at the epidermal-dermal junction. In atypical nevi, features such as lentiginous hyperplasia, acanthosis and architectural disorder were imaged. Cytological atypia was present in all the melanoma lesions imaged, showing the strongest correlation with malignancy. The MPM images demonstrated very good correlation with corresponding histological images, suggesting that MPM could be a promising tool for in vivo non-invasive pigmented lesion diagnosis, particularly distinguishing atypical nevi from melanoma.

Highlights

  • The current standard for melanoma detection is based on clinical visual assessment of the lesions (ABCDE rule) [1], biopsy and histopathology

  • The multiphoton microscopy (MPM) features of melanocytic nevi were characterized by normal morphology of keratinocytes of the epidermal layers, nests of nevus cells surrounded by collagen fibers at the epidermaldermal junction (EDJ) and elongated rete ridges compared to normal skin

  • Melanocytic nevi lesions were not biopsied, but the features identified by the MPM imaging were in good correlation with features generally identified by histopathology in compound nevi (Figure 2a)

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Summary

Introduction

The current standard for melanoma detection is based on clinical visual assessment of the lesions (ABCDE rule) [1], biopsy and histopathology. The ABCD approach is effective in distinguishing well between the two extreme stages of melanocytic lesions: common nevi and melanoma. Considered a gold standard for diagnosing melanocytic nevi, this routine diagnostic method is affected by limitations in discriminating early melanoma from atypical nevi for a large number of lesions that fall into the borderline area. This creates the problem of false-negative diagnosis that could delay diagnosis and treatment, and of false-positive diagnosis, which could lead to unnecessary biopsies and treatments. Non-invasive optical imaging technologies based on laser-scanning microscopy have emerged as promising tools for real time, in situ imaging of skin lesions with the potential, if clinically accepted, to reduce the amount of excision and specimen processing as in conventional histopathology. [2,3,4]

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