Coordinate-targeted fluorescence nanoscopy with multiple off states

Danzl, Johann G.; Sidenstein, Sven C.; Gregor, Carola; Urban, Nicolai T.; Ilgen, Peter; Jakobs, Stefan; Hell, Stefan W.

doi:10.1038/nphoton.2015.266

Cited by 84 publications

(66 citation statements)

References 28 publications

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“…Resolution enhancement along all three dimensions would be helpful for a more detailed characterization of the complex 3D structure of synaptic sites. Furthermore, the presented setup allows the implementation of continuous-wave beams needed for reversible saturable optical fluorescence transition (RESOLFT)31 and Protected STED32 nanoscopy with reversibly switchable EGFP variants33.…”

Section: Discussionmentioning

confidence: 99%

Multicolour Multilevel STED nanoscopy of Actin/Spectrin Organization at Synapses

Sidenstein

D’Este

Böhm

et al. 2016

Sci Rep

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108

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Superresolution fluorescence microscopy of multiple fluorophores still requires development. Here we present simultaneous three-colour stimulated emission depletion (STED) nanoscopy relying on a single STED beam at 620 nm. Toggling the STED beam between two or more power levels (“multilevelSTED”) optimizes resolution and contrast in all colour channels, which are intrinsically co-aligned and well separated. Three-colour recording is demonstrated by imaging the nanoscale cytoskeletal organization in cultured hippocampal neurons. The down to ~35 nm resolution identified periodic actin/betaII spectrin lattices along dendrites and spines; however, at presynaptic and postsynaptic sites, these patterns were found to be absent. Both our multicolour scheme and the 620 nm STED line should be attractive for routine STED microscopy applications.

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Section: Discussionmentioning

confidence: 99%

Multicolour Multilevel STED nanoscopy of Actin/Spectrin Organization at Synapses

Sidenstein

D’Este

Böhm

et al. 2016

Sci Rep

Self Cite

108

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“…As another approach, we have recently proposed structurally reinforced p-conjugated skeletons that contain an electron-withdrawing phosphine oxide (P=O) moiety. [7] Thei ncorporation of aP =Og roup also leads to bathochromic shifts of the absorption and emission maxima. [7] Thei ncorporation of aP =Og roup also leads to bathochromic shifts of the absorption and emission maxima.…”

mentioning

confidence: 99%

A Highly Photostable Near‐Infrared Labeling Agent Based on a Phospha‐rhodamine for Long‐Term and Deep Imaging

et al. 2018

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Various fluorescence microscopy techniques require bright NIR-emitting fluorophores with high chemical and photostability. Now, the significant performance improvement of phosphorus-substituted rhodamine dyes (PORs) upon substitution at the 9-position with a 2,6-dimethoxyphenyl group is reported. The thus obtained dye PREX 710 was used to stain mitochondria in living cells, which allowed long-term and three-color imaging in the vis-NIR range. Moreover, the high fluorescence longevity of PREX 710 allows tracking a dye-labeled biomolecule by single-molecule microscopy under physiological conditions. Deep imaging of blood vessels in mice brain has also been achieved using the bright NIR-emitting PREX 710-dextran conjugate.

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“…Stimulated emission depletion (STED) microscopy is a powerful imaging technique that can break the diffraction limit . STED microscopy acquires super‐resolution images by superimposing a donut‐shaped depletion beam on the excitation beam spot of confocal microscopy, selectively deactivating the fluorescence emission.…”

Section: Introductionmentioning

confidence: 99%

Enhanced lateral resolution in continuous wave stimulated emission depletion microscopy using tightly focused annular radially polarized excitation beam

Lim

Kim

et al. 2019

Journal of Biophotonics

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The lateral resolution of continuous wave (CW) stimulated emission depletion (STED) microscopy is enhanced about 12% by applying annular‐shaped amplitude modulation to the radially polarized excitation beam. A focused annularly filtered radially polarized excitation beam provides a more condensed point spread function (PSF), which contributes to enhance effective STED resolution of CW STED microscopy. Theoretical analysis shows that the FWHM of the effective PSF on the detection plane is smaller than for conventional CW STED. Simulation shows the donut‐shaped PSF of the depletion beam and confocal optics suppress undesired PSF sidelobes. Imaging experiments agree with the simulated resolution improvement.

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Coordinate-targeted fluorescence nanoscopy with multiple off states

Cited by 84 publications

References 28 publications

Multicolour Multilevel STED nanoscopy of Actin/Spectrin Organization at Synapses

Multicolour Multilevel STED nanoscopy of Actin/Spectrin Organization at Synapses

A Highly Photostable Near‐Infrared Labeling Agent Based on a Phospha‐rhodamine for Long‐Term and Deep Imaging

Enhanced lateral resolution in continuous wave stimulated emission depletion microscopy using tightly focused annular radially polarized excitation beam

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