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. 2017 Apr 19;7:951. doi: 10.1038/s41598-017-01122-7

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© The Author(s) 2017

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Spectral crosstalk between actin rings and vesicles labelled by Alexa 647 and Alexa 568 dyes, respectively, using a single multiband filter set and excitation of 647 nm (a,c) and 561 nm (b,d). Additional thresholding of the accepted localizations based on their photon number can reduce the crosstalk (c,d). Emission spectra of Alexa 568 (orange) and 647 (red) filtered by the multiband emission filter (black). Scale bar is 1 micron.