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Research Letter |

New Diagnostic Method for Lesions With Transepidermal Melanocytic Migration

Matthias Scholz, MDSc1; Susanne Buder, MD2; Katrin Kerl, MD3; Reinhard Dummer, MD3; Claus Garbe, MD4
[+] Author Affiliations
1LTB Lasertechnik Berlin, Berlin, Germany
2Vivantes Klinikum Neukölln, Klinik für Dermatologie und Venerologie, Berlin, Germany
3Department of Dermatology, University Hospital of Zurich, Zurich, Switzerland
4Zentrum für Dermatoonkologie,Universität Tübingen, Universitäts-Hautklinik, Tübingen, Germany
JAMA Dermatol. 2014;150(6):654-656. doi:10.1001/jamadermatol.2013.8119.
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Transepidermal melanocytic migration (TEM) is a histological feature that is frequently observed in malignant melanoma but may also occur in nevi (eg, Spitz nevi, acral nevi). Pagetoid TEM is considered a key indicator of malignant disease. We refer to an unusual case report by Kerl et al1 in a young female patient. Eleven melanomas were originally diagnosed within 10 years, and in a comprehensive retrospective diagnosis, these could be reclassified as nevi. (See Kerl et al1 for details of the patient’s medical history and the context of this case.) Herein, we report on the application of a new diagnostic method for melanocytic lesions to the issue of TEM as melanoma indicator.

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Figure 1.
Schematic Representation of Different Types of Fluorescence Excitation of Organic Fluorophores (Simplified)

The figure shows a Jablonski diagram. The length of the arrow indicates the energy of the photon (wavelength is written next to the arrow). The solid arrows indicate laser photons, and the dashed arrows indicate fluorescence photons. A, Conventional 1-photon excitation; B, simultaneous 2-photon excitation; C, stepwise 2-photon excitation of melanin fluorescence. A and B illustrate excitation suitable for measuring the conventional autofluorescence of skin tissue. The corresponding fluorophores show no absorption of 800-nm photons; therefore, the latter can be used for fluorescence excitation only in a simultaneous 2-photon absorption process. However, melanin has sufficient absorption at 800 nm and therefore can absorb 2 such photons in a stepwise absorption process (mechanism illustrated in panel C), which is substantially more effective than the mechanism shown in panel B. This way, the ultraweak fluorescence of melanin in skin, which otherwise is completely masked by the autofluorescence, is detectable. (For details, see Eichhorn et al.3)

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Figure 2.
Characteristics of Stepwise 2-Photon Excited Melanin Fluorescence Spectra

A-C, Paraffin-embedded skin tissue samples (measured with 800-nm/2.5-ns pulses). A, Normally pigmented skin; B, nevus; and C, malignant melanocytic melanoma. Each tissue sample was measured at histologically proven reference samples, especially in panel B, a compound nevus, and in panel C, a superficially spreading melanoma. D and E, Ascending cells in the upper epidermis. D, In all of the 8 paraffin-embedded samples of the present study (E) in a histologically proven, paraffin-embedded sample of a nodular melanoma with transepidermal melanocytic migration (measured with 800 nm/2.5 ns pulses). The measurement points are integral intensity values via 16 nm (error bars indicate means [SDs] of measurements in 10 areas of the tissue type). The diameter of the measured skin area is 50 µm. The solid lines are for guidance only. An additional signal appears at 400 nm, if the measuring area is in the dermis, resulting from collagen. Under the present conditions of excitation, there is no contribution of paraffin to the fluorescence.

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