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Fig. 2. | BMC Biology

Fig. 2.

From: Q&A: Expansion microscopy

Fig. 2.

Nanoscale imaging with ExM. ac Imaging at 60–70 nm resolution of mouse hippocampus after viral delivery of membrane-bound Brainbow3.0 followed by antibody staining, using protein retention ExM (proExM) and imaged on a confocal microscope. a Maximum intensity projection (MIP) of an image stack. b Pre-expansion image showing one optical section of the boxed region in a. c Post-expansion image of b. d, e Nanoscale imaging of mouse brain using proExM with post-expansion antibody delivery. d Post-expansion image of mouse cortex from Thy1-YFP mouse after high-temperature treatment, followed by immunostaining against bassoon (blue), homer (red), and YFP (green). e Post-expansion image of mouse cortex from Thy1-YFP mouse after high-temperature treatment, followed by immunostaining against myelin basic protein (red) and YFP (green). f, g Nanoscale imaging of RNA in Thy1–YFP mouse brain using expansion fluorescence in situ hybridization (ExFISH). f Wide-field image showing YFP protein (red), and hybridization chain reaction ExFISH (HCR-ExFISH) signals for YFP mRNA (cyan) and Gad1 mRNA (magenta). g Confocal image of mouse hippocampus from f. Inset: one plane of the boxed region. Colors as in f. hj Imaging at ~25 nm resolution of mouse hippocampus using iterative expansion microscopy (iExM). h Image of unexpanded mouse hippocampus after viral delivery of membrane-bound Brainbow3.0, followed by antibody staining against EYFP (blue), TagBFP (red), and mTFP (green). i As in h, but expanded using proExM. Inset: a magnified image of spines from the dotted box of i. j MIP of an image stack of iExM-expanded mouse hippocampus with staining against EYFP (blue) and mCherry (green). Inset: zoomed-out view of the image of j. Scale bars, in biological units: a 50 μm (physical size post-expansion 198 μm); b 5 μm; c 5 μm (19.8 μm); d 5 μm (21 μm); e 5 μm (21 μm); f 500 μm (1450 μm); g 50 μm (145 μm), inset 10 μm (29 μm); h 3 μm; i 3 μm (14 μm), inset 1 μm (4.5 μm); j 1 μm (20 μm), inset 3 μm (60 μm). Images adapted by permission from Macmillan Publishers Ltd: Nature Biotechnology [4], copyright 2016; Nature Methods [5], copyright 2016; Nature Methods [6], copyright 2017

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