article published yes Mario Perkovic author Michael Kunz author Ulrike Endesfelder author Stefanie Bunse author Christoph Wigge author Zhou Yu author Victor-Valentin Hodirnau author 3661B498-F248-11E8-B48F-1D18A9856A87 Margot P. Scheffer author Anja Seybert author Sebastian Malkusch author Erin M. Schuman author Mike Heilemann author Achilleas S. Frangakis author Correlative microscopy incorporates the specificity of fluorescent protein labeling into high-resolution electron micrographs. Several approaches exist for correlative microscopy, most of which have used the green fluorescent protein (GFP) as the label for light microscopy. Here we use chemical tagging and synthetic fluorophores instead, in order to achieve protein-specific labeling, and to perform multicolor imaging. We show that synthetic fluorophores preserve their post-embedding fluorescence in the presence of uranyl acetate. Post-embedding fluorescence is of such quality that the specimen can be prepared with identical protocols for scanning electron microscopy (SEM) and transmission electron microscopy (TEM); this is particularly valuable when singular or otherwise difficult samples are examined. We show that synthetic fluorophores give bright, well-resolved signals in super-resolution light microscopy, enabling us to superimpose light microscopic images with a precision of up to 25 nm in the x–y plane on electron micrographs. To exemplify the preservation quality of our new method we visualize the molecular arrangement of cadherins in adherens junctions of mouse epithelial cells. https://research-explorer.ista.ac.at/download/9655/9701/2014_JournalOfStructuralBiology_Perkovic.pdf application/pdfno https://creativecommons.org/licenses/by-nc-nd/3.0/ Elsevier2014 eng 1047-8477 2469895410.1016/j.jsb.2014.03.018 1862205-213 yes Perkovic M, Kunz M, Endesfelder U, Bunse S, Wigge C, Yu Z, Hodirnau V-V, Scheffer MP, Seybert A, Malkusch S, Schuman EM, Heilemann M, Frangakis AS. 2014. Correlative light- and electron microscopy with chemical tags. Journal of Structural Biology. 186(2), 205–213. Perkovic, M., Kunz, M., Endesfelder, U., Bunse, S., Wigge, C., Yu, Z., … Frangakis, A. S. (2014). Correlative light- and electron microscopy with chemical tags. <i>Journal of Structural Biology</i>. Elsevier. <a href="https://doi.org/10.1016/j.jsb.2014.03.018">https://doi.org/10.1016/j.jsb.2014.03.018</a> Perkovic, Mario, Michael Kunz, Ulrike Endesfelder, Stefanie Bunse, Christoph Wigge, Zhou Yu, Victor-Valentin Hodirnau, et al. “Correlative Light- and Electron Microscopy with Chemical Tags.” <i>Journal of Structural Biology</i>. Elsevier, 2014. <a href="https://doi.org/10.1016/j.jsb.2014.03.018">https://doi.org/10.1016/j.jsb.2014.03.018</a>. Perkovic, Mario, et al. “Correlative Light- and Electron Microscopy with Chemical Tags.” <i>Journal of Structural Biology</i>, vol. 186, no. 2, Elsevier, 2014, pp. 205–13, doi:<a href="https://doi.org/10.1016/j.jsb.2014.03.018">10.1016/j.jsb.2014.03.018</a>. M. Perkovic <i>et al.</i>, “Correlative light- and electron microscopy with chemical tags,” <i>Journal of Structural Biology</i>, vol. 186, no. 2. Elsevier, pp. 205–213, 2014. Perkovic M, Kunz M, Endesfelder U, et al. Correlative light- and electron microscopy with chemical tags. <i>Journal of Structural Biology</i>. 2014;186(2):205-213. doi:<a href="https://doi.org/10.1016/j.jsb.2014.03.018">10.1016/j.jsb.2014.03.018</a> M. Perkovic, M. Kunz, U. Endesfelder, S. Bunse, C. Wigge, Z. Yu, V.-V. Hodirnau, M.P. Scheffer, A. Seybert, S. Malkusch, E.M. Schuman, M. Heilemann, A.S. Frangakis, Journal of Structural Biology 186 (2014) 205–213. 96552021-07-14T09:05:42Z2021-07-22T08:26:32Z