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. 2020 Dec 14;10:21842. doi: 10.1038/s41598-020-78827-9

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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TIRF microscopy imaging with fluorescent ligand 10b. TIRF images of CHO cells stably expressing (a) D_2SR or (b) D₃R labeled with 10b (10 nM for D_2SR, 1 nM for D₃R) visualize the receptor distribution on the cell surface and demonstrate the suitability of the fluorescent ligand for high resolution fluorescence microscopy. (c, d) Validation of specific labeling: representative brightfield (left) and TIRF images (right) of a CHO cell stably expressing (c) D_2SR or (d) D₃R, preincubated with the D_2/3R antagonist spiperone (10 μM) and treated with 10b (10 nM) show only a few non-specifically adhered, immobile fluorescent spots and no cell-specific labeling.