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EMBO Rep. 2018 Sep;19(9). pii: e45836. doi: 10.15252/embr.201845836. Epub 2018 Jul 9.

X10 expansion microscopy enables 25-nm resolution on conventional microscopes.

Author information

1
Institute for Neuro- and Sensory Physiology, Center for Biostructural Imaging of Neurodegeneration, Cluster of Excellence Nanoscale Microscopy and Molecular Physiology of the Brain, University Medical Center Göttingen, Göttingen, Germany strucke@gwdg.de srizzol@gwdg.de.
2
International Max Planck Research School for Molecular Biology, Göttingen, Germany.
3
Institute for Neuro- and Sensory Physiology, Center for Biostructural Imaging of Neurodegeneration, Cluster of Excellence Nanoscale Microscopy and Molecular Physiology of the Brain, University Medical Center Göttingen, Göttingen, Germany.

Abstract

Expansion microscopy is a recently introduced imaging technique that achieves super-resolution through physically expanding the specimen by ~4×, after embedding into a swellable gel. The resolution attained is, correspondingly, approximately fourfold better than the diffraction limit, or ~70 nm. This is a major improvement over conventional microscopy, but still lags behind modern STED or STORM setups, whose resolution can reach 20-30 nm. We addressed this issue here by introducing an improved gel recipe that enables an expansion factor of ~10× in each dimension, which corresponds to an expansion of the sample volume by more than 1,000-fold. Our protocol, which we termed X10 microscopy, achieves a resolution of 25-30 nm on conventional epifluorescence microscopes. X10 provides multi-color images similar or even superior to those produced with more challenging methods, such as STED, STORM, and iterative expansion microscopy (iExM). X10 is therefore the cheapest and easiest option for high-quality super-resolution imaging currently available. X10 should be usable in any laboratory, irrespective of the machinery owned or of the technical knowledge.

KEYWORDS:

STED ; STORM ; expansion microscopy; nanoscopy; super‐resolution

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