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Two-photon excitation STED microscopy
Open accessAbstract
We report sub-diffraction resolution in two-photon excitation (TPE) fluorescence microscopy achieved by merging this technique with stimulated-emission depletion (STED). We demonstrate an easy-to-implement and promising laser combination based on a short-pulse laser source for two-photon excitation and a continuous-wave (CW) laser source for resolution enhancement. Images of fluorescent nanoparticles and the immunostained transcription regulator NF kappaB in mammalian cell nuclei exhibit resolutions of <50 nm and approximately 70 nm in the focal plane, respectively, corresponding to a 4-5.4-fold improvement over the diffraction barrier.
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“The STED principle can also be applied to two-photon microscopy (Moneron and Hell, 2009).”
From Barriers in the brain: resolving dendritic spine morphology and compartmentalization (2014) · cites this paper as Moneron and Hell, 2009
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Metadata
- Authors
- Moneron, G., & Hell, S. W.
- ORCID
- Hell, Stefan W.
- Venue
- Optics Express
- Publisher
- Optica Publishing Group
- Published
- 2009-08-03
- Volume
- 17
- Issue
- 17
- Pages
- 14567
- ISSN
- 1094-4087 (ISSN-L)
- OpenAlex
- W2152298085
- Cited by
- 238
- References
- 21
- Citations by year
- 2026: 7 · 2025: 4 · 2024: 6 · 2023: 10 · 2022: 11 · 2021: 11 · 2020: 12 · 2019: 15 · 2018: 19 · 2017: 18 · 2016: 15 · 2015: 18 · 2014: 25 · 2013: 18 · 2012: 18
- FWCI
- 9.49
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Optics Express (publishedVersion)
- PubMed (publishedVersion)
- Max Planck Institute for Plasma Physics (submittedVersion)
- GRO (submittedVersion)
- MPG.PuRe (Max Planck Society) (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Physical Sciences › Physics and Astronomy › Atomic and Molecular Physics, and Optics › Laser-Matter Interactions and Applications
- Physical Sciences › Materials Science › Electronic, Optical and Magnetic Materials › Nonlinear Optical Materials Studies
- Keywords
- two-photon excitation, STED microscopy, stimulated emission depletion, sub-diffraction resolution, super-resolution fluorescence microscopy, short-pulse lasers, continuous-wave laser, fluorescent nanoparticles, NF-κB, immunostaining
- MeSH terms
- Animals, Cell Nucleus, Fluorescent Dyes, Humans, Lasers, Microscopy, Electron, Scanning, Microscopy, Fluorescence, Models, Statistical, NF-kappa B, Photons, Microscopy, Confocal, Imaging, Three-Dimensional, Nanotechnology, Nanoparticles
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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