ArticleBiophysical journal2013
Live-cell superresolution imaging by pulsed STED two-photon excitation microscopy.
Article in Biophysical journal, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 49 papers.
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Who cites it
49 citing papers in PubMed.
- Wavelength-tunable depletion expands fluorophore compatibility in two-photon STED nanoscopy.Biomedical optics express · 2026Article
- Contribution of individual excitatory synapses on dendritic spines to electrical signaling.Frontiers in neuroscience · 2025Article
- Correlative multiphoton-STED microscopy of podocyte calcium levels and slit diaphragm ultrastructure in the renal glomerulus.Scientific reports · 2024Article
- Three-Dimensional Microfluidic Capillary Device for Rapid and Multiplexed Immunoassays in Whole Blood.ACS sensors · 2024Article
- A pipeline for STED super-resolution imaging and Imaris analysis of nanoscale synapse organization in mouse cortical brain slices.STAR protocols · 2023Article
- [Frontiers and development in live-cell super-resolution fluorescence microscopy].Sheng wu yi xue gong cheng xue za zhi = Journal of biomedical engineering = Shengwu yixue gongchengxue zazhi · 2023Review
- Article
- Improving two-photon excitation microscopy for sharper and faster biological imaging.Biophysics and physicobiology · 2023Article
- Commercially derived versatile optical architecture for two-photon STED, wavelength mixing and label-free microscopy.Biomedical optics express · 2022Article
- Aberration correction in stimulated emission depletion microscopy to increase imaging depth in living brain tissue.Neurophotonics · 2021Article
- High spatial-resolution imaging of label-freeThe Analyst · 2021Article
- Article
- Between life and death: strategies to reduce phototoxicity in super-resolution microscopy.Journal of physics D: Applied physics · 2020Review
- Current Techniques for Investigating the Brain Extracellular Space.Frontiers in neuroscience · 2020Review
- Efficient two-photon excitation stimulated emission depletion nanoscope exploiting spatiotemporal information.Neurophotonics · 2019Article
- Two-photon STED nanoscopy realizing 100-nm spatial resolution utilizing high-peak-power sub-nanosecond 655-nm pulses.Biomedical optics express · 2019Article
- Article
- Full volume super-resolution imaging of thick mitotic spindle using 3D AO STED microscope.Biomedical optics express · 2019Article
- Unveiling the Extracellular Space of the Brain: From Super-resolved Microstructure toThe Journal of neuroscience : the official journal of the Society for Neuroscience · 2018Review
- Multi-photon near-infrared emission saturation nanoscopy using upconversion nanoparticles.Nature communications · 2018Article
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3 authors.
Funding
Abstract
Two-photon laser scanning microscopy (2PLSM) allows fluorescence imaging in thick biological samples where absorption and scattering typically degrade resolution and signal collection of one-photon imaging approaches. The spatial resolution of conventional 2PLSM is limited by diffraction, and the near-infrared wavelengths used for excitation in 2PLSM preclude the accurate imaging of many small subcellular compartments of neurons. Stimulated emission depletion (STED) microscopy is a superresolution imaging modality that overcomes the resolution limit imposed by diffraction and allows fluorescence imaging of nanoscale features. Here, we describe the design and operation of a superresolution two-photon microscope using pulsed excitation and STED lasers. We examine the depth dependence of STED imaging in acute tissue slices and find enhancement of 2P resolution ranging from approximately fivefold at 20 μm to approximately twofold at 90-μm deep. The depth dependence of resolution is found to be consistent with the depth dependence of depletion efficiency, suggesting resolution is limited by STED laser propagation through turbid tissue. Finally, we achieve live imaging of dendritic spines with 60-nm resolution and demonstrate that our technique allows accurate quantification of neuronal morphology up to 30-μm deep in living brain tissue.
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