Hell, Stefan W.

[["dc.bibliographiccitation.firstpage","11440"],["dc.bibliographiccitation.issue","31"],["dc.bibliographiccitation.journal","Proceedings of the National Academy of Sciences of the United States of America"],["dc.bibliographiccitation.lastpage","11445"],["dc.bibliographiccitation.volume","103"],["dc.contributor.author","Donnert, Gerald"],["dc.contributor.author","Keller, Jan"],["dc.contributor.author","Medda, Rebecca"],["dc.contributor.author","Andrei, M. Alexandra"],["dc.contributor.author","Rizzoli, Silvio"],["dc.contributor.author","Lührmann, Reinhard"],["dc.contributor.author","Jahn, Reinhard"],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2017-09-07T11:52:39Z"],["dc.date.available","2017-09-07T11:52:39Z"],["dc.date.issued","2006"],["dc.description.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 approximate to 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."],["dc.identifier.doi","10.1073/pnas.0604965103"],["dc.identifier.gro","3143651"],["dc.identifier.isi","000239616400005"],["dc.identifier.pmid","16864773"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/1188"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","0027-8424"],["dc.title","Macromolecular-scale resolution in biological fluorescence microscopy"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","143"],["dc.bibliographiccitation.issue","3"],["dc.bibliographiccitation.journal","Journal of Microscopy"],["dc.bibliographiccitation.lastpage","148"],["dc.bibliographiccitation.volume","184"],["dc.contributor.author","Schrader, M."],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2022-03-01T11:47:10Z"],["dc.date.available","2022-03-01T11:47:10Z"],["dc.date.issued","2003"],["dc.identifier.doi","10.1111/jmi.1996.184.3.143"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/103937"],["dc.language.iso","en"],["dc.notes.intern","DOI-Import GROB-531"],["dc.relation.issn","0022-2720"],["dc.title","Wavefronts in the focus of a light microscope"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","144"],["dc.bibliographiccitation.issue","3"],["dc.bibliographiccitation.journal","Nature Photonics"],["dc.bibliographiccitation.lastpage","147"],["dc.bibliographiccitation.volume","3"],["dc.contributor.author","Rittweger, Eva"],["dc.contributor.author","Han, Kyu Young"],["dc.contributor.author","Irvine, Scott E."],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2017-09-07T11:47:33Z"],["dc.date.available","2017-09-07T11:47:33Z"],["dc.date.issued","2009"],["dc.description.abstract","Because they have spin states that can be optically polarized and detected, fluorescent nitrogen vacancies in diamond(1-3) have considerable potential for applications in quantum cryptography(4,5) and computation(6-8), as well as for nanoscale magnetic imaging(9,10) and biolabelling(11,12). However, their optical detection and control are hampered by the diffraction resolution barrier of far-field optics. Here, we show that stimulated emission depletion (STED) microscopy(13,14) is capable of imaging nitrogen-vacancy centres with nanoscale resolution and Angstrom precision using focused light. The far-field optical control of the population of their excited state at the nanoscale expands the versatility of these centres and demonstrates the suitability of STED microscopy to image dense colour centres in crystals. Nitrogen-vacancy defects show great potential as tags for far-field optical nanoscopy(15) because they exhibit nearly ideal STED without bleaching. Measured point-spread functions of 5.8 nm in width demonstrate an all-physics-based far-field optical resolving power exceeding the wavelength of light by two orders of magnitude."],["dc.identifier.doi","10.1038/NPHOTON.2009.2"],["dc.identifier.gro","3143147"],["dc.identifier.isi","000264289600014"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/629"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","1749-4885"],["dc.title","STED microscopy reveals crystal colour centres with nanometric resolution"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","210a"],["dc.bibliographiccitation.issue","2, Supplement 1"],["dc.bibliographiccitation.journal","Biophysical Journal"],["dc.bibliographiccitation.lastpage","211a"],["dc.bibliographiccitation.volume","104"],["dc.contributor.author","Sitters, Gerrit"],["dc.contributor.author","Heller, Iddo"],["dc.contributor.author","Broekmans, Onno D."],["dc.contributor.author","Farge, Géraldine A."],["dc.contributor.author","Menges, Carolin"],["dc.contributor.author","Wende, Wolfgang"],["dc.contributor.author","Hell, Stefan"],["dc.contributor.author","Peterman, Erwin J. G."],["dc.contributor.author","Wuite, Gijs J. L."],["dc.date.accessioned","2017-09-07T11:53:10Z"],["dc.date.available","2017-09-07T11:53:10Z"],["dc.date.issued","2013"],["dc.identifier.doi","10.1016/j.bpj.2012.11.1190"],["dc.identifier.gro","3145041"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/2733"],["dc.language.iso","en"],["dc.notes.intern","Crossref Import"],["dc.notes.status","final"],["dc.relation.issn","0006-3495"],["dc.title","STED Nanoscopy Combined with Optical Tweezers Reveals Spatial Dynamics of Proteins on Densely Covered DNA"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dc.type.peerReviewed","no"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","RP1"],["dc.bibliographiccitation.issue","2"],["dc.bibliographiccitation.journal","J. Microsc."],["dc.bibliographiccitation.lastpage","RP2"],["dc.bibliographiccitation.volume","180"],["dc.contributor.author","Hell, S. W."],["dc.contributor.author","Schrader, M."],["dc.contributor.author","Bahlmann, K."],["dc.contributor.author","Meinecke, F."],["dc.contributor.author","Lakowicz, J. R ."],["dc.contributor.author","Gryczynski, I."],["dc.date.accessioned","2018-03-21T14:33:54Z"],["dc.date.available","2018-03-21T14:33:54Z"],["dc.date.issued","1995"],["dc.description.abstract","We report the realization of stimulated emission on a microscopic scale. An experiment is presented describing significant depopulation of the excited state of the fluorophore Pyridine 2 with a mode‐locked Ti: Sapphire laser. Stimulated emission is performed at 750 nm and excitation at the frequency doubled wavelength of 375 nm. The pulses are synchronized so that the stimulating pulse follows the excitation pulse after 5 ps. The set‐up is a modified 4Pi‐confocal microscope employing one of the objective lenses for excitation and the opposing one for stimulated emission."],["dc.identifier.doi","10.1111/j.1365-2818.1995.tb03662.x"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/13111"],["dc.language.iso","en"],["dc.notes.status","final"],["dc.notes.status","final"],["dc.title","Stimulated Emission on a Microscopic Scale: Light Quenching of Pyridinium 2 using a Ti:Sapphire laser"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","207"],["dc.bibliographiccitation.issue","3"],["dc.bibliographiccitation.journal","Single Molecules"],["dc.bibliographiccitation.lastpage","210"],["dc.bibliographiccitation.volume","2"],["dc.contributor.author","Hell, Stefan W."],["dc.contributor.author","Mar Blanca, Carlo"],["dc.date.accessioned","2017-09-07T11:46:43Z"],["dc.date.available","2017-09-07T11:46:43Z"],["dc.date.issued","2001"],["dc.description.abstract","A major goal of improving the resolution in far-field light microscopy is to create observation volumes that are sharp and spherical. Two-photon excitation 4Pi-confocal fluorescence microscopy provides a sharp and nearly spherical focal diffraction maximum, but the maximum is accompanied by axial side-lobes of 30-40% relative height. Whereas in 4Pi-confocal imaging the effect of the lobes on the image can be eliminated by data processing, in spectroscopy their presence contributes to the observation volume in full. Here we show that simple binary amplitude filters, placed in the entrance aperture of novel 1.45 numerical aperture oil immersion lenses, suppress the side-lobes below 7%. As a result, a nearly spherical sharp observation volume is created consisting of a solitary spot of similar to150 nm full-width-half-maximum."],["dc.identifier.doi","10.1002/1438-5171(200110)2:3<207::aid-simo207>3.0.co;2-s"],["dc.identifier.gro","3144331"],["dc.identifier.isi","000174074600007"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/1943"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","1438-5163"],["dc.relation.issn","1438-5163"],["dc.title","Sharp Spherical Focal Spot by Dark Ring 4Pi-Confocal Microscopy"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.artnumber","042004"],["dc.bibliographiccitation.issue","4"],["dc.bibliographiccitation.journal","Methods and Applications in Fluorescence"],["dc.bibliographiccitation.volume","3"],["dc.contributor.author","Sednev, Maksim V."],["dc.contributor.author","Belov, Vladimir N."],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2017-09-07T11:54:49Z"],["dc.date.available","2017-09-07T11:54:49Z"],["dc.date.issued","2015"],["dc.description.abstract","The review deals with commercially available organic dyes possessing large Stokes shifts and their applications as fluorescent labels in optical microscopy based on stimulated emission depletion (STED). STED microscopy breaks Abbe's diffraction barrier and provides optical resolution beyond the diffraction limit. STED microscopy is non-invasive and requires photostable fluorescent markers attached to biomolecules or other objects of interest. Up to now, in most biology-related STED experiments, bright and photoresistant dyes with small Stokes shifts of 20-40 nm were used. The rapid progress in STED microscopy showed that organic fluorophores possessing large Stokes shifts are indispensable in multi-color super-resolution techniques. The ultimate result of the imaging relies on the optimal combination of a dye, the bio-conjugation procedure and the performance of the optical microscope. Modern bioconjugation methods, basics of STED microscopy, as well as structures and spectral properties of the presently available fluorescent markers are reviewed and discussed. In particular, the spectral properties of the commercial dyes are tabulated and correlated with the available depletion wavelengths found in STED microscopes produced by LEICA Microsytems, Abberior Instruments and Picoquant GmbH."],["dc.identifier.doi","10.1088/2050-6120/3/4/042004"],["dc.identifier.gro","3141768"],["dc.identifier.isi","000380688100007"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/857"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","2050-6120"],["dc.title","Fluorescent dyes with large Stokes shifts for super-resolution optical microscopy of biological objects: a review"],["dc.type","review"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","915"],["dc.bibliographiccitation.issue","11"],["dc.bibliographiccitation.journal","Nature Methods"],["dc.bibliographiccitation.lastpage","918"],["dc.bibliographiccitation.volume","4"],["dc.contributor.author","Willig, Katrin I."],["dc.contributor.author","Harke, Benjamin"],["dc.contributor.author","Medda, Rebecca"],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2017-09-07T11:49:24Z"],["dc.date.available","2017-09-07T11:49:24Z"],["dc.date.issued","2007"],["dc.description.abstract","We report stimulated emission depletion (STED) fluorescence microscopy with continuous wave (CW) laser beams. Lateral fluorescence confinement from the scanning focal spot delivered a resolution of 29 - 60 nm in the focal plane, corresponding to a 5 - 8- fold improvement over the diffraction barrier. Axial spot confinement increased the axial resolution by 3.5-fold. We observed three-dimensional (3D) subdiffraction resolution in 3D image stacks. Viable for fluorophores with low triplet yield, the use of CW light sources greatly simplifies the implementation of this concept of far-field fluorescence nanoscopy."],["dc.identifier.doi","10.1038/NMETH1108"],["dc.identifier.gro","3143419"],["dc.identifier.isi","000250575700014"],["dc.identifier.pmid","17952088"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/931"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","1548-7091"],["dc.title","STED microscopy with continuous wave beams"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","49"],["dc.bibliographiccitation.issue","1"],["dc.bibliographiccitation.journal","Journal of the Optical Society of America A"],["dc.bibliographiccitation.lastpage","54"],["dc.bibliographiccitation.volume","18"],["dc.contributor.author","Nagorni, Matthias"],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2017-09-07T11:46:42Z"],["dc.date.available","2017-09-07T11:46:42Z"],["dc.date.issued","2001"],["dc.description.abstract","We analyze the ability of nonlinear image restoration to remove interference artifacts in microscopes that enlarge the axial optical bandwidth through coherent counterpropagating waves. We calculate the images of an elaborate test object as produced by confocal, standing-wave, incoherent illumination interference image interference, and 4Pi confocal microscopes, and we subsequently investigate the extent to which the initial object can be restored by the information allowed by their optical transfer function. We find that nonlinear restoration is successful only if the transfer function is sufficiently contiguous and has amplitudes well above the noise level, as is mostly the case in a two-photon excitation 4Pi confocal microscope."],["dc.identifier.doi","10.1364/JOSAA.18.000049"],["dc.identifier.gro","3144327"],["dc.identifier.isi","000166042500004"],["dc.identifier.pmid","11152003"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/1939"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","0740-3232"],["dc.title","Coherent use of opposing lenses for axial resolution increase. II. Power and limitation of nonlinear image restoration"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","337"],["dc.bibliographiccitation.issue","2"],["dc.bibliographiccitation.journal","European Journal of Organic Chemistry"],["dc.bibliographiccitation.lastpage","349"],["dc.bibliographiccitation.volume","2015"],["dc.contributor.author","Mitronova, Gyuzel Yu"],["dc.contributor.author","Polyakova, Svetlana"],["dc.contributor.author","Wurm, Christian Andreas"],["dc.contributor.author","Kolmakov, Kirill"],["dc.contributor.author","Wolfram, Thomas"],["dc.contributor.author","Meineke, Dirk N. H."],["dc.contributor.author","Belov, Vladimir N."],["dc.contributor.author","John, Michael"],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2017-09-07T11:44:46Z"],["dc.date.available","2017-09-07T11:44:46Z"],["dc.date.issued","2015"],["dc.description.abstract","Aromatic nucleophilic substitution (SNAr) of fluorine in 9-(3'-carboxy-4',5',6',7'-tetrafluorophenyl) groups of xanthene dyes constitutes a powerful tool in dye design. Thiols and amines regioselectively replace F-6'. This approach enables additional hydrophilic residues or functional groups required for bioconjugation to be introduced. By using this methodology, a \"bright\" and photostable dye for two-color superresolution microscopy was synthesized (with absorption and emission maxima at 604 and 627 nm, respectively). In the case of red-emitting rhodamine dyes with 3'-carboxy-4',5',7'-trifluorophenyl residues, two-dimensional NMR techniques and a chemical transformation were used to prove the precise position of the additional substituent - a carboxylic acid group linked through the S-atom at C-6'. Furthermore, simple H-1 NMR spectra reliably permit the position of the additional carboxy substituent in the 3'-carboxyphenyl ring (at C-5' or C-6') to be established. Information on the exact position of this substituent is significant for the design of molecular probes and for the prediction of the properties of their bioconjugates."],["dc.identifier.doi","10.1002/ejoc.201403269"],["dc.identifier.gro","3141988"],["dc.identifier.isi","000347722900011"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/3301"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.eissn","1099-0690"],["dc.relation.issn","1434-193X"],["dc.title","Functionalization of the meso-Phenyl Ring of Rhodamine Dyes Through SNAr with Sulfur Nucleophiles: Synthesis, Biophysical Characterizations, and Comprehensive NMR Analysis"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]