ArticleProceedings of the National Academy of Sciences of the United States of America2006
Macromolecular-scale resolution in biological fluorescence microscopy.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 118 papers.
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118 citing papers in PubMed, 527 citations in OpenAlex.
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- Dense 4D nanoscale reconstruction of living brain tissue.Nature methods · 2023Article
- Deep learning enables fast, gentle STED microscopy.Communications biology · 2023Article
- Advanced Microscopy Techniques for Molecular Biophysics.International journal of molecular sciences · 2023Review
- Far-field super-resolution chemical microscopy.Light, science & applications · 2023Review
- Deep learning enables fast, gentle STED microscopy.bioRxiv : the preprint server for biology · 2023Article
- Chiral and nematic phases of flexible active filaments.Nature physics · 2023Article
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- Multimodal imaging of synaptic vesicles with a single probe.Cell reports methods · 2022Article
- Investigation of Galectins in Frozen Tissue and Mammalian Cell Culture Using Confocal Miccroscopy.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Long-term STED imaging of membrane packing and dynamics by exchangeable polarity-sensitive dyes.Biophysical reports · 2021Article
- [Comparison and progress review of various super-resolution fluorescence imaging techniques].Se pu = Chinese journal of chromatography · 2021Review
- Aberration correction in stimulated emission depletion microscopy to increase imaging depth in living brain tissue.Neurophotonics · 2021Article
58 more citing papers are in PubMed but not listed here.
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Authors and funding
9 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We demonstrate far-field fluorescence microscopy with a focal-plane resolution of 15-20 nm in biological samples. The 10- to 12-fold multilateral increase in resolution below the diffraction barrier has been enabled by the elimination of molecular triplet state excitation as a major source of photobleaching of a number of dyes in stimulated emission depletion microscopy. Allowing for relaxation of the triplet state between subsequent excitation-depletion cycles yields an up to 30-fold increase in total fluorescence signal as compared with reported stimulated emission depletion illumination schemes. Moreover, it enables the reduction of the effective focal spot area by up to approximately 140-fold below that given by diffraction. Triplet-state relaxation can be realized either by reducing the repetition rate of pulsed lasers or by increasing the scanning speed such that the build-up of the triplet state is effectively prevented. This resolution in immunofluorescence imaging is evidenced by revealing nanoscale protein patterns on endosomes, the punctuated structures of intermediate filaments in neurons, and nuclear protein speckles in mammalian cells with conventional optics. The reported performance of diffraction-unlimited fluorescence microscopy opens up a pathway for addressing fundamental problems in the life sciences.
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