Scientific article
Open access

Molecular resolution imaging by post-labeling expansion single-molecule localization microscopy (Ex-SMLM)

Published inNature Communications, vol. 11, no. 1, p. 1-11; 3388
Publication date2020-07-07
First online date2020-07-07

Expansion microscopy (ExM) enables super-resolution fluorescence imaging of physically expanded biological samples with conventional microscopes. By combining ExM with single-molecule localization microscopy (SMLM) it is potentially possible to approach the resolution of electron microscopy. However, current attempts to combine both methods remained challenging because of protein and fluorophore loss during digestion or denaturation, gelation, and the incompatibility of expanded polyelectrolyte hydrogels with photoswitching buffers. Here we show that re-embedding of expanded hydrogels enables d STORM imaging of expanded samples and demonstrate that post-labeling ExM resolves the current limitations of super-resolution microscopy. Using microtubules as a reference structure and centrioles, we demonstrate that post-labeling Ex-SMLM preserves ultrastructural details, improves the labeling efficiency and reduces the positional error arising from linking fluorophores into the gel thus paving the way for super-resolution imaging of immunolabeled endogenous proteins with true molecular resolution.

Citation (ISO format)
ZWETTLER, Fabian U. et al. Molecular resolution imaging by post-labeling expansion single-molecule localization microscopy (Ex-SMLM). In: Nature Communications, 2020, vol. 11, n° 1, p. 1–11. doi: 10.1038/s41467-020-17086-8
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Article (Published version)
ISSN of the journal2041-1723

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