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. 2020 Mar 13;12:1758835920905415. doi: 10.1177/1758835920905415

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© The Author(s), 2020

This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage).

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Figure 4. — Detection of circulating tumor cells (CTCs) in melanoma patients. (a) shows representative patterns of CTCs isolated from a patient with metastatic melanoma (#349). As shown, nuclei were not damaged (Hoechst-33342 positive cells) while all isolated populations were negative for the expression of CD45, CD31 and CD34. Line 1 reveals PE⁺ CTCs (CD44⁺ or CD146⁺ or NCAD⁺), line 2 the positivity for the FITC-conjugated monoclonal antibodies (CD271 or ABCB5 or RANK), line 3 the double positivity (PE⁺/FITC⁺) and line 4 the CTCs negative for all the mentioned antigens. The bottom panel in (a) shows the negative control represented by lymphocytes detected by the expression of CD45 or CD31 or CD34. (b) shows the phenotypic profile of CTCs isolated from the 17 patients with metastatic melanoma. (c) is representative of the mutational assessment by Droplet Digital polymerase chain reaction (ddPCR) of CTCs from two patients previously investigated for the presence of BRAF^V600E mutation in primary tumor. The ddPCR analysis produced the same results obtained in primary melanoma specimens.