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","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","1651"],["dc.bibliographiccitation.issue","7"],["dc.bibliographiccitation.journal","Biophysical Journal"],["dc.bibliographiccitation.lastpage","1660"],["dc.bibliographiccitation.volume","101"],["dc.contributor.author","Mueller, Veronica"],["dc.contributor.author","Ringemann, Christian"],["dc.contributor.author","Honigmann, Alf"],["dc.contributor.author","Schwarzmann, Guenter"],["dc.contributor.author","Medda, Rebecca"],["dc.contributor.author","Leuschner, Ivo"],["dc.contributor.author","Polyakova, Svetlana"],["dc.contributor.author","Belov, Vladimir N."],["dc.contributor.author","Hell, Stefan"],["dc.contributor.author","Eggeling, Christian"],["dc.date.accessioned","2017-09-07T11:43:23Z"],["dc.date.available","2017-09-07T11:43:23Z"],["dc.date.issued","2011"],["dc.description.abstract","Details about molecular membrane dynamics in living cells, such as lipid-protein interactions, are often hidden from the observer because of the limited spatial resolution of conventional far-field optical microscopy. The superior spatial resolution of stimulated emission depletion (STED) nanoscopy can provide new insights into this process. The application of fluorescence correlation spectroscopy (FCS) in focal spots continuously tuned down to 30 nm in diameter distinguishes between free and anomalous molecular diffusion due to, for example, transient binding of lipids to other membrane constituents, such as lipids and proteins. We compared STED-FCS data recorded on various fluorescent lipid analogs in the plasma membrane of living mammalian cells. Our results demonstrate details about the observed transient formation of molecular complexes. The diffusion characteristics of phosphoglycerolipids without hydroxyl-containing headgroups revealed weak interactions. The strongest interactions were observed with sphingolipid analogs, which showed cholesterol-assisted and cytoskeleton-dependent binding. The hydroxyl-containing headgroup of gangliosides, galactosylceramide, and phosphoinositol assisted binding, but in a much less cholesterol- and cytoskeleton-dependent manner. The observed anomalous diffusion indicates lipid-specific transient hydrogen bonding to other membrane molecules, such as proteins, and points to a distinct connectivity of the various lipids to other membrane constituents. This strong interaction is different from that responsible for forming cholesterol-dependent, liquid-ordered domains in model membranes."],["dc.identifier.doi","10.1016/j.bpj.2011.09.006"],["dc.identifier.gro","3142651"],["dc.identifier.isi","000295661300011"],["dc.identifier.pmid","21961591"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/78"],["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","0006-3495"],["dc.title","STED Nanoscopy Reveals Molecular Details of Cholesterol- and Cytoskeleton-Modulated Lipid Interactions in Living Cells"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","571"],["dc.bibliographiccitation.issue","7"],["dc.bibliographiccitation.journal","Nature Methods"],["dc.bibliographiccitation.lastpage","573"],["dc.bibliographiccitation.volume","8"],["dc.contributor.author","Vicidomini, Giuseppe"],["dc.contributor.author","Moneron, Gael"],["dc.contributor.author","Han, Kyu Young"],["dc.contributor.author","Westphal, Volker"],["dc.contributor.author","Ta, Haisen"],["dc.contributor.author","Reuss, Matthias"],["dc.contributor.author","Engelhardt, Johann"],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2017-09-07T11:44:10Z"],["dc.date.available","2017-09-07T11:44:10Z"],["dc.date.issued","2011"],["dc.description.abstract","Applying pulsed excitation together with time-gated detection improves the fluorescence on-off contrast in continuous-wave stimulated emission depletion (CW-STED) microscopy, thus revealing finer details in fixed and living cells using moderate light intensities. This method also enables super-resolution fluorescence correlation spectroscopy with CW-STED beams, as demonstrated by quantifying the dynamics of labeled lipid molecules in the plasma membrane of living cells."],["dc.identifier.doi","10.1038/NMETH.1624"],["dc.identifier.gro","3142707"],["dc.identifier.isi","000292194500020"],["dc.identifier.pmid","21642963"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/141"],["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","Sharper low-power STED nanoscopy by time gating"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","178"],["dc.bibliographiccitation.issue","2"],["dc.bibliographiccitation.journal","Quarterly Reviews of Biophysics"],["dc.bibliographiccitation.lastpage","243"],["dc.bibliographiccitation.volume","48"],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Willig, Katrin I."],["dc.contributor.author","Sahl, Steffen J."],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2017-09-07T11:44:24Z"],["dc.date.available","2017-09-07T11:44:24Z"],["dc.date.issued","2015"],["dc.description.abstract","The majority of studies of the living cell rely on capturing images using fluorescence microscopy. Unfortunately, for centuries, diffraction of light was limiting the spatial resolution in the optical microscope: structural and molecular details much finer than about half the wavelength of visible light (similar to 200nm) could not be visualized, imposing significant limitations on this otherwise so promising method. The surpassing of this resolution limit in far-field microscopy is currently one of the most momentous developments for studying the living cell, as the move from microscopy to super-resolution microscopy or \"nanoscopy' offers opportunities to study problems in biophysical and biomedical research at a new level of detail. This review describes the principles and modalities of present fluorescence nanoscopes, as well as their potential for biophysical and cellular experiments. All the existing nanoscopy variants separate neighboring features by transiently preparing their fluorescent molecules in states of different emission characteristics in order to make the features discernible. Usually these are fluorescent 'on' and 'off' states causing the adjacent molecules to emit sequentially in time. Each of the variants can in principle reach molecular spatial resolution and has its own advantages and disadvantages. Some require specific transitions and states that can be found only in certain fluorophore subfamilies, such as photoswitchable fluorophores, while other variants can be realized with standard fluorescent labels. Similar to conventional far-field microscopy, nanoscopy can be utilized for dynamical, multi-color and three-dimensional imaging of fixed and live cells, tissues or organisms. Lens-based fluorescence nanoscopy is poised for a high impact on future developments in the life sciences, with the potential to help solve long-standing quests in different areas of scientific research."],["dc.identifier.doi","10.1017/S0033583514000146"],["dc.identifier.gro","3141910"],["dc.identifier.isi","000354386800002"],["dc.identifier.pmid","25998828"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/2433"],["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","1469-8994"],["dc.relation.issn","0033-5835"],["dc.title","Lens-based fluorescence nanoscopy"],["dc.type","review"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.journal","Molecular Biology of the Cell"],["dc.bibliographiccitation.volume","25"],["dc.contributor.author","Saka, S. K."],["dc.contributor.author","Honigmann, A."],["dc.contributor.author","Eggeling, C."],["dc.contributor.author","Hell, Stefan W."],["dc.contributor.author","Lang, T."],["dc.contributor.author","Rizzoli, Silvio"],["dc.date.accessioned","2018-11-07T09:31:35Z"],["dc.date.available","2018-11-07T09:31:35Z"],["dc.date.issued","2014"],["dc.identifier.isi","000352094103168"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/31561"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Amer Soc Cell Biology"],["dc.publisher.place","Bethesda"],["dc.relation.eventlocation","Philadelphia, PA"],["dc.relation.issn","1939-4586"],["dc.relation.issn","1059-1524"],["dc.title","Multi-protein assemblies underlie the mesoscale organization of the plasma membrane."],["dc.type","conference_abstract"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","6266"],["dc.bibliographiccitation.issue","33"],["dc.bibliographiccitation.journal","Angewandte Chemie International Edition"],["dc.bibliographiccitation.lastpage","6270"],["dc.bibliographiccitation.volume","46"],["dc.contributor.author","Fölling, Jonas"],["dc.contributor.author","Belov, Vladimir N."],["dc.contributor.author","Kunetsky, R."],["dc.contributor.author","Medda, Rebecca"],["dc.contributor.author","Schoenle, Andreas"],["dc.contributor.author","Egner, Alexander"],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Bossi, Mariano L."],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2017-09-07T11:49:52Z"],["dc.date.available","2017-09-07T11:49:52Z"],["dc.date.issued","2007"],["dc.description.abstract","Exciting developments: Switching individual photochromic and fluorescent rhodamine amides enables 3D far-field optical microscopy with nanoscale resolution, excellent signal-to-noise ratio, and fast acquisition times. The rhodamine amides can be switched on using two photons, which enables 3D detailed imaging of thick and densely stained samples (such as 5-μm silica beads (see image) and living cells) to be constructed."],["dc.identifier.doi","10.1002/anie.200702167"],["dc.identifier.gro","3143552"],["dc.identifier.isi","000249114700006"],["dc.identifier.pmid","17640007"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/1078"],["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","1433-7851"],["dc.title","Photochromic rhodamines provide nanoscopy with optical sectioning"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","634a"],["dc.bibliographiccitation.issue","2, Supplement 1"],["dc.bibliographiccitation.journal","Biophysical Journal"],["dc.bibliographiccitation.lastpage","634a"],["dc.bibliographiccitation.volume","106"],["dc.contributor.author","Honigmann, Alf"],["dc.contributor.author","Sadeghi, Sina"],["dc.contributor.author","Jan, Keller"],["dc.contributor.author","Hell, Stefan"],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Vink, Richard"],["dc.date.accessioned","2017-09-07T11:53:03Z"],["dc.date.available","2017-09-07T11:53:03Z"],["dc.date.issued","2014"],["dc.description.abstract","The cytoplasmic side of eukaryotic cell membranes is covered by a dense actin rich cortex. We present FCS and STED experiments showing that a dense membrane bound actin network has severe influence on temperature dependent lipid phase separation. A minimal actin cortex was bound to a supported lipid bilayer via biotinylated lipid streptavidin complexes (pinning sites). In general, actin binding to ternary membranes prevented macroscopic liquid-ordered (Lo) and liquid-disordered (Ld) domain formation in these systems, even at low temperature. For pinning sites that strongly attract Ld domains, an actin correlated multi-domain pattern was observed, consisting of Ld “channels” along the actin fibers, with Lo “islands” in the voids. FCS measurements revealed enhanced diffusion of unsaturated lipids along the channels, and hindered diffusion of these lipids in directions perpendicular. For pinning sites strongly attractive to Lo domains, an “inverse” domain structure was observed. These findings are in good agreement with a number of recently proposed simulation models. However, to fully capture our experimental observations, an extended simulation model is proposed, in which the lipid domains also couple to the local membrane curvature. Our results provide a mechanism how cells may prevent macroscopic de-mixing of membrane components and at the same time regulate the local membrane compositions."],["dc.identifier.doi","10.1016/j.bpj.2013.11.3506"],["dc.identifier.gro","3145018"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/2708"],["dc.language.iso","en"],["dc.notes.intern","Crossref Import"],["dc.notes.status","final"],["dc.relation.issn","0006-3495"],["dc.title","A Lipid Bound Actin Meshwork Organizes Liquid Phase Separation in Model Membranes"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dc.type.peerReviewed","no"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.artnumber","e01671"],["dc.bibliographiccitation.journal","eLife"],["dc.bibliographiccitation.volume","3"],["dc.contributor.author","Honigmann, Alf"],["dc.contributor.author","Sadeghi, Sina"],["dc.contributor.author","Keller, Jan"],["dc.contributor.author","Hell, Stefan"],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Vink, Richard"],["dc.date.accessioned","2017-09-07T11:46:25Z"],["dc.date.available","2017-09-07T11:46:25Z"],["dc.date.issued","2014"],["dc.description.abstract","The eukaryotic cell membrane is connected to a dense actin rich cortex. We present FCS and STED experiments showing that dense membrane bound actin networks have severe influence on lipid phase separation. A minimal actin cortex was bound to a supported lipid bilayer via biotinylated lipid streptavidin complexes (pinning sites). In general, actin binding to ternary membranes prevented macroscopic liquid-ordered and liquid-disordered domain formation, even at low temperature. Instead, depending on the type of pinning lipid, an actin correlated multi-domain pattern was observed. FCS measurements revealed hindered diffusion of lipids in the presence of an actin network. To explain our experimental findings, a new simulation model is proposed, in which the membrane composition, the membrane curvature, and the actin pinning sites are all coupled. Our results reveal a mechanism how cells may prevent macroscopic demixing of their membrane components, while at the same time regulate the local membrane composition."],["dc.identifier.doi","10.7554/eLife.01671"],["dc.identifier.fs","605546"],["dc.identifier.gro","3142167"],["dc.identifier.isi","000333192500001"],["dc.identifier.pmid","24642407"],["dc.identifier.purl","https://resolver.sub.uni-goettingen.de/purl?gs-1/10516"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/5277"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.intern","Merged from goescholar"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.publisher","Elife Sciences Publications Ltd"],["dc.relation.issn","2050-084X"],["dc.relation.orgunit","Fakultät für Physik"],["dc.rights","CC BY 3.0"],["dc.title","A lipid bound actin meshwork organizes liquid phase separation in model membranes"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dc.type.version","published_version"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.artnumber","7368"],["dc.bibliographiccitation.firstpage","204"],["dc.bibliographiccitation.journal","Nature"],["dc.bibliographiccitation.lastpage","208"],["dc.bibliographiccitation.volume","478"],["dc.contributor.author","Grotjohann, Tim"],["dc.contributor.author","Testa, Ilaria"],["dc.contributor.author","Leutenegger, Marcel"],["dc.contributor.author","Bock, Hannes"],["dc.contributor.author","Urban, Nicolai T."],["dc.contributor.author","Lavoie-Cardinal, Flavie"],["dc.contributor.author","Willig, Katrin I."],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Jakobs, Stefan"],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2017-09-07T11:43:21Z"],["dc.date.available","2017-09-07T11:43:21Z"],["dc.date.issued","2011"],["dc.description.abstract","Lens-based optical microscopy failed to discern fluorescent features closer than 200 nm for decades, but the recent breaking of the diffraction resolution barrier by sequentially switching the fluorescence capability of adjacent features on and off is making nanoscale imaging routine. Reported fluorescence nanoscopy variants switch these features either with intense beams at defined positions or randomly, molecule by molecule. Here we demonstrate an optical nanoscopy that records raw data images from living cells and tissues with low levels of light. This advance has been facilitated by the generation of reversibly switchable enhanced green fluorescent protein (rsEGFP), a fluorescent protein that can be reversibly photoswitched more than a thousand times. Distributions of functional rsEGFP-fusion proteins in living bacteria and mammalian cells are imaged at <40-nanometre resolution. Dendritic spines in living brain slices are super-resolved with about a million times lower light intensities than before. The reversible switching also enables all-optical writing of features with subdiffraction size and spacings, which can be used for data storage."],["dc.identifier.doi","10.1038/nature10497"],["dc.identifier.gro","3142644"],["dc.identifier.isi","000295782800041"],["dc.identifier.pmid","21909116"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/71"],["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","0028-0836"],["dc.title","Diffraction-unlimited all-optical imaging and writing with a photochromic GFP"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]