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Journal Article

Expansion Stimulated Emission Depletion Microscopy (ExSTED).

MPS-Authors
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Gao,  Mengfei
Max Planck Institute for Molecular Cell Biology and Genetics, Max Planck Society;

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Maraspini,  Riccardo
Max Planck Institute for Molecular Cell Biology and Genetics, Max Planck Society;

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Beutel,  Oliver
Max Planck Institute for Molecular Cell Biology and Genetics, Max Planck Society;

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Honigmann,  Alf
Max Planck Institute for Molecular Cell Biology and Genetics, Max Planck Society;

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Citation

Gao, M., Maraspini, R., Beutel, O., Zehtabian, A., Eickholt, B., Honigmann, A., et al. (2018). Expansion Stimulated Emission Depletion Microscopy (ExSTED). ACS nano, 12(5), 4178-4185. doi:10.1021/acsnano.8b00776.


Cite as: https://hdl.handle.net/21.11116/0000-0003-F5EB-0
Abstract
Stimulated emission depletion (STED) microscopy is routinely used to resolve the ultrastructure of cells with a ∼10-fold higher resolution compared to diffraction limited imaging. While STED microscopy is based on preparing the excited state of fluorescent probes with light, the recently developed expansion microscopy (ExM) provides subdiffraction resolution by physically enlarging the sample before microscopy. The expansion of the fixed cells by cross-linking and swelling of hydrogels easily enlarges the sample ∼4-fold and hence increases the effective optical resolution by this factor. To overcome the current limits of these complementary approaches, we combined ExM with STED (ExSTED) and demonstrated an increase in resolution of up to 30-fold compared to conventional microscopy (<10 nm lateral and ∼50 nm isotropic). While the increase in resolution is straightforward, we found that high-fidelity labeling via multi-epitopes is required to obtain emitter densities that allow ultrastructural details with ExSTED to be resolved. Our work provides a robust template for super-resolution microscopy of entire cells in the ten nanometer range.