Journal
SCIENCE ADVANCES
Volume 8, Issue 23, Pages -Publisher
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/sciadv.abo2506
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Funding
- European Research Council (ERC) via project smMIET under the European Union [884488]
- Germany's Excellence Strategy [EXC 2067/1-390729940]
- DFG [GRK 2157]
- European Research Council via the ERC Synergy Grant project ULTRARESOLUTION [951275]
- Intramural Research Program of the National Institutes of Health, National Cancer Institute, Center for Cancer Research [BC011506]
- Open Access Publication Funds of the Gottingen University
- European Research Council (ERC) [951275, 884488] Funding Source: European Research Council (ERC)
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The MIET imaging method utilizes metal-induced energy transfer to achieve nanometer-level axial resolution, and combines it with the excellent lateral resolution of dSTORM to enable three-dimensional super-resolution imaging of subcellular structures.
Over the past two decades, super-resolution microscopy has seen a tremendous development in speed and resolution, but for most of its methods, there exists a remarkable gap between lateral and axial resolution, which is by a factor of 2 to 3 worse. One recently developed method to close this gap is metal-induced energy transfer (MIET) imaging, which achieves an axial resolution down to nanometers. It exploits the distance-dependent quenching of fluorescence when a fluorescent molecule is brought close to a metal surface. In the present manuscript, we combine the extreme axial resolution of MIET imaging with the extraordinary lateral resolution of single-molecule localization microscopy, in particular with direct stochastic optical reconstruction microscopy (dSTORM). This combination allows us to achieve isotropic three-dimensional super-resolution imaging of subcellular structures. Moreover, we used spectral demixing for implementing dual-color MIET-dSTORM that allows us to image and colocalize, in three dimensions, two different cellular structures simultaneously.
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