Fig. 16

Combining ExM and super-resolution microscopy methods. a–e Expansion STED (ExSTED). a Magnified view of the microtubules, showing clear distinction of the antibody positions on the opposing sites of a single microtubule. Scale bar, 100 nm. b Quantification of the apparent diameter of microtubule and the 2D resolution. c 3D STED of the expanded microtubules using an oil objective. Scale bar, 500 nm. d Model of the centrioles (blue) and the centriole-associated protein CEP152. e ExSTED HeLa cells with CEP152 staining. Scale bar, 200 nm. f–h Expansion SIM (ExSIM). f Sectioning of the expanded sample. g Z-projection of the reconstructed SIM image. h By matching the RI of the immersion oil with the sample, fine details of the Drosophila SC structure can be analyzed. Scale bars, 5 μm. i–o Expansion SMLM (ExSMLM). i dSTORM image of pre-labeled, proExM-expanded and re-embedded Cos-7 cells stained with primary antibodies against α-tubulin and Alexa Fluor 532-conjugated secondary antibodies. Scale bar, 500 nm. j 3D dSTORM image of the expanded and re-embedded Cos-7 cells labeled with anti α- and β-tubulin antibodies and additionally post-labeling with anti-α-tubulin. Scale bar, 5 μm. k Magnified view of highlighted region (white box) in j. Scale bar, 500 nm. l Image projection of the xz-axes averaged along two microtubule filaments shown in j. Scale bar, 100 nm. m Cross-sectional profile (blue dots) of the xz-projection shown in l. Using a bi-Gaussian fit (red), the peak-to-peak distance is determined to 81.2 nm. n, o 3D dSTORM image of U-ExM expanded and re-embedded Chlamydomonas centrioles stained post re-embedding. Zoom-in image of the boxed area in n is shown in o. Scale bar, 1 μm (n), 500 nm (o).   (a–e) Reprinted/adapted with permission from Gao et al. [97]. Copyright 2018 American Chemical Society. f–h Reprinted/adapted by permission from Springer Nature: Wang et al. [99], copyright 2018. i–o Reprinted/adapted from Zwettler et al. [43]