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. 2022 Apr 5;13:1842. doi: 10.1038/s41467-022-29546-4

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© The Author(s) 2022

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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Fig. 3 — 55% sucrose was added to the buffer to slow down the protein diffusion by 30× and ease observing the autofluorescence bursts. a Autofluorescence time traces with increasing β-galactosidase-streptavidin concentrations. The binning time is 30 ms. The stars indicate selected autofluorescence bursts which are displayed on the right panel with 15 ms bin time to better view individual bursts. b Normalized photon counts histograms computed over the full trace duration of 200 s. The thick lines are numerical fits using the sum of a Gaussian and an exponential distribution to account for the noise and the autofluorescence bursts respectively. The inset shows the number of detected events per minute above the threshold corresponding to 2.5× the standard deviation of the noise when no protein is present (dashed lines at 310 counts in (a). c FCS correlation functions of the traces displayed in (a). The thick lines are numerical fits.