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Fig. 15 | Nano Convergence

Fig. 15

From: Nanoscale fluorescence imaging of biological ultrastructure via molecular anchoring and physical expansion

Fig. 15

(a, b Reprinted/adapted from Damstra et al. [48]. c–e Reprinted/adapted from Truckenbrodt et al. [49]. f, g Reprinted/adapted from M’Saad et al. [47]. ho Reprinted/adapted by permission from Rockefeller University Press: ©2021 Shi et al. [55]. Originally published in J. Cell. Biol. https://doi.org/10.1083/jcb.202105067. p Reprinted/adapted from Sarkar et al. [52] by permission from the author)

High-resolution images obtained with high-expansion-factor ExM variants. a Nucleus imaged by confocal microscopy after TREx. Scale bar, 1 μm. b High-resolution view of several nuclear pores from boxed region (2) of a and the distribution of diameters of individual nuclear pores. Scale bar, 200 nm. c Immunostainings for the peroxisome membrane protein Pmp70 in neurons by both super-resolution and X10 Expansion Microscopy. Scale bar, 100 nm. d The exemplary line scan from the X10 Expansion Microscopy image in c is shown with a best Gaussian fit curve. e Quantification of the average resolution of X10 Expansion Microscopy image, which is 25.2 ± 0.2 nm. f pan-ExM reveals nuclear architecture in interphase. Image of SYTOX Green nucleic acid stain (middle column) and NHS ester pan-stained sample (left column) and overlay of them (right column). Scale bars, (top row) 5 μm, (middle row) 250 nm, (bottom row), 1 μm. g Overlay of the NHS ester and anti-GFP images. The insets show the zoom-in yellow boxes and reveal individual ManII-positive Golgi cisternae. Scale bars, 2 μm. h Label-retention expansion STORM (LR-ExSTORM) image of a HeLa cell overexpressing SNAP-CLTB, stained with BG-MA-biotin, and post expansion labeled with streptavidin-AF647. Scale bar, 2 μm. i, j Images of x–y cross sections at the top of single CCPs as illustrated in m. Scale bar, 100 nm. k, l Images of x–y cross sections in the middle of single CCPs as illustrated in n. Images in il are different CCPs. Scale bar, 100 nm. o Nearest cluster distance analysis of 134 CCPs imaged with LR-STORM. p Expansion Revealing (ExR) reveals co-localized clusters of Aβ42 peptide and potassium ion channels in the fornix of Alzheimer’s model 5× FAD mice. The image showing post-expansion Aβ42 (magenta), SMI (cyan) and Kv7.2 (yellow) staining in the fornix of a 5xFAD mouse (Scale bar, 400 nm). The left most panel, merged low magnification image (Scale bar, 4 μm).

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