Nanoscale subcellular architecture revealed by multicolor 3D salvaged fluorescence imaging

Combining the molecular specificity of fluorescent probes with three-dimensional (3D) imaging at nanoscale resolution is critical for investigating the spatial organization and interactions of cellular organelles and protein complexes. We present a super-resolution light microscope that enables simu...

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Published inNature methods Vol. 17; no. 2; pp. 225 - 231
Main Authors Zhang, Yongdeng, Schroeder, Lena K., Lessard, Mark D., Kidd, Phylicia, Chung, Jeeyun, Song, Yuanbin, Benedetti, Lorena, Li, Yiming, Ries, Jonas, Grimm, Jonathan B., Lavis, Luke D., De Camilli, Pietro, Rothman, James E., Baddeley, David, Bewersdorf, Joerg
Format Journal Article
LanguageEnglish
Published 06.01.2020
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Summary:Combining the molecular specificity of fluorescent probes with three-dimensional (3D) imaging at nanoscale resolution is critical for investigating the spatial organization and interactions of cellular organelles and protein complexes. We present a super-resolution light microscope that enables simultaneous multicolor imaging of mammalian cells at 5–10 nm single-molecule localization precision in 3D. We show its power for cell biology research with fluorescence images that resolve the highly convoluted Golgi apparatus and the close contacts between the endoplasmic reticulum and the plasma membrane, structures that have traditionally been the imaging realm of electron microscopy.
Bibliography:Y.Z. and J.B. conceived the idea. Y.Z. implemented the hardware. Y.Z., Y.L., J.R., and D.B. wrote the software. L.K.S., J.C., P.D.C., J.E.R., and J.B. designed biological experiments. L.K.S. developed sample preparation protocols. L.K.S. prepared and optimized microtubule, ER, mitochondria, Golgi, and ER-PM contact sites samples. Y.S. and L.B. provided the synaptonemal complex samples. M.D.L. prepared and optimized the synaptonemal complex samples. P.K. prepared and optimized microtubule samples. Y.Z. imaged the samples and analyzed the images. J.B.G. and L.D.L. synthesized PA-JF646 and PA-JF669. Y.Z., L.K.S., and J.B. wrote the manuscript with input from all authors. All authors discussed the results and commented on the manuscript.
Author contributions
ISSN:1548-7091
1548-7105
DOI:10.1038/s41592-019-0676-4