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.
“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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- 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 › Engineering › Biomedical Engineering › 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
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