Fluorescent Rhodamines and Fluorogenic Carbopyronines for Super‐Resolution STED Microscopy in Living Cells

Butkevich, Alexey N.; Mitronova, Gyuzel Yu.; Sidenstein, Sven C.; Klocke, Jessica L.; Kamin, Dirk; Meineke, Dirk N. H.; D’Este, Elisa; Kraemer, Philip Tobias; Danzl, Johann G.; Belov, Vladimir N.; Hell, Stefan W.

doi:10.1002/anie.201511018

Cited by 217 publications

(265 citation statements)

References 42 publications

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“…Therefore, we tested a water-soluble 6 ′ -carboxy-Q -rhodamine19 ( 540R ) absorbing and emitting at 540 nm and 561 nm, respectively. This dye possesses a compact, zwitterionic structure with a relatively short distance between positive and negative charges, which is characteristic of cell-penetrating synthetic probes20. After incubation with its Halo-tag amine conjugate ( 540R -Halo, Supplementary Appendix 1), living HeLa cells expressing Halo-tag fusion proteins showed bright staining with low background and permitted 620 nm STED imaging with a resolution of ~45 nm (Supplementary Fig.…”

Section: Resultsmentioning

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: Resultsmentioning

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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“…Among the multitude of fluorophores reported so far, only rhodamines, carbopyronines, and silicon‐rhodamines (SiR) bearing a carboxyl in the ortho ‐position of the pendant aromatic ring provide specific vital labeling and perform well in super‐resolution fluorescence microscopy. These dyes exist in equilibrium between zwitterionic (fluorescent) and spirolactone (non‐fluorescent) forms.…”

Section: Figurementioning

confidence: 99%

Hydroxylated Fluorescent Dyes for Live‐Cell Labeling: Synthesis, Spectra and Super‐Resolution STED

Butkevich

Belov

Kolmakov

et al. 2017

Chemistry A European J

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106

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Hydroxylated rhodamines, carbopyronines, silico‐ and germanorhodamines with absorption maxima in the range of 530–640 nm were prepared and applied in specific labeling of living cells. The direct and high‐yielding entry to germa‐ and silaxanthones tolerates the presence of protected heteroatoms and may be considered for the syntheses of various sila‐ and germafluoresceins, as well as ‐rhodols. Application in stimulated emission depletion (STED) fluorescence microscopy revealed a resolution of 50–75 nm in one‐ and two‐color imaging of vimentin‐HaloTag fused protein and native tubulin. The established structure–property relationships allow for prediction of the spectral properties and the positions of spirolactone/zwitterion equilibria for the new analogues of rhodamines, carbo‐, silico‐, and germanorhodamines using simple additive schemes.

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“…. In combination with novel fluorogenic probes, such as SiR and 580R , that can be imaged with the 775 nm STED laser line, the live‐cell potential for STED nanoscopy is increased. STORM and PALM have been performed with fusion proteins or organic fluorophores that have intrinsic blinking properties or can be activated by light.…”

Section: Super‐resolution Techniques For Virus Imagingmentioning

confidence: 99%

Super‐resolved insights into human immunodeficiency virus biology

et al. 2016

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Edited by Wilhelm JustThe recent development of fluorescence microscopy approaches overcoming the diffraction limit of light microscopy opened possibilities for studying small-scale cellular processes. The spatial resolution achieved by these novel techniques, together with the possibility to perform live-cell and multicolor imaging, make them ideally suited for visualization of native viruses and subviral structures within the complex environment of a host cell or organ, thus providing fundamentally new possibilities for investigating virus-cell interactions. Here, we review the use of super-resolution microscopy approaches to study virus-cell interactions, and discuss recent insights into human immunodeficiency virus biology obtained by exploiting these novel techniques.

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Fluorescent Rhodamines and Fluorogenic Carbopyronines for Super‐Resolution STED Microscopy in Living Cells

Cited by 217 publications

References 42 publications

Multicolour Multilevel STED nanoscopy of Actin/Spectrin Organization at Synapses

Multicolour Multilevel STED nanoscopy of Actin/Spectrin Organization at Synapses

Hydroxylated Fluorescent Dyes for Live‐Cell Labeling: Synthesis, Spectra and Super‐Resolution STED

Super‐resolved insights into human immunodeficiency virus biology

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