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. 2021 Dec 9;221(2):e202110137. doi: 10.1083/jcb.202110137

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© 2021 Paramasivam et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure 4. — Multicolor SMLM detects and visualizes singular LNP-mRNA in endosomal compartments with nanometer resolution. (A–D) Exemplary images of endosomes containing a single, clearly detectable LNP-Cy5-mRNA (magenta) together with transferrin (green) and EGF (cyan) cargo for each imaged LNP formulation (top row). SMLM resolves single LNP as round nano domains (zoom-in; middle row) with characteristic size distribution. LNP diameter distributions were built by determining the FWHM of single LNPs (see Materials and methods). Mean LNP diameters (FWHM) were calculated by Gaussian fitting of the distributions to (mean ± SD) 74.9 ± 22.6, 71.0 ± 22.0, 66.7 ± 20.2, and 68.9 ± 21.7 nm, respectively. The cellular context of displayed endosomes is provided in Supplemental figures, Figs. S10, S25, S26, and S27. Scale bars are 100 nm. rel., relative.