Willig, Katrin I.

[["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"]]

[["dc.bibliographiccitation.firstpage","756"],["dc.bibliographiccitation.issue","4"],["dc.bibliographiccitation.journal","ChemPhysChem"],["dc.bibliographiccitation.lastpage","762"],["dc.bibliographiccitation.volume","15"],["dc.contributor.author","Jensen, Nickels A."],["dc.contributor.author","Danzl, Johann G."],["dc.contributor.author","Willig, Katrin I."],["dc.contributor.author","Lavoie-Cardinal, Flavie"],["dc.contributor.author","Brakemann, Tanja"],["dc.contributor.author","Hell, Stefan W."],["dc.contributor.author","Jakobs, Stefan"],["dc.date.accessioned","2017-09-07T11:46:25Z"],["dc.date.available","2017-09-07T11:46:25Z"],["dc.date.issued","2014"],["dc.description.abstract","Diffraction-unlimited far-field super-resolution fluorescence (nanoscopy) methods typically rely on transiently transferring fluorophores between two states, whereby this transfer is usually laid out as a switch. However, depending on whether this is induced in a spatially controlled manner using a pattern of light (coordinate-targeted) or stochastically on a single-molecule basis, specific requirements on the fluorophores are imposed. Therefore, the fluorophores are usually utilized just for one class of methods only. In this study we demonstrate that the reversibly switchable fluorescent protein Dreiklang enables live-cell recordings in both spatially controlled and stochastic modes. We show that the Dreiklang chromophore entails three different light-induced switching mechanisms, namely a reversible photochemical one, off-switching by stimulated emission, and a reversible transfer to a long-lived dark state from the S-1 state, all of which can be utilized to overcome the diffraction barrier. We also find that for the single-molecule-based stochastic GSDIM approach (ground-state depletion followed by individual molecule return), Dreiklang provides a larger number of on-off localization events as compared to its progenitor Citrine. Altogether, Dreiklang is a versatile probe for essentially all popular forms of live-cell fluorescence nanoscopy."],["dc.identifier.doi","10.1002/cphc.201301034"],["dc.identifier.gro","3142169"],["dc.identifier.isi","000332747500026"],["dc.identifier.pmid","24497300"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/5299"],["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","1439-7641"],["dc.relation.issn","1439-4235"],["dc.title","Coordinate-Targeted and Coordinate-Stochastic Super-Resolution Microscopy with the Reversibly Switchable Fluorescent Protein Dreiklang"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","992"],["dc.bibliographiccitation.issue","6"],["dc.bibliographiccitation.journal","Neuron"],["dc.bibliographiccitation.lastpage","1000"],["dc.bibliographiccitation.volume","75"],["dc.contributor.author","Testa, Ilaria"],["dc.contributor.author","Urban, Nicolai T."],["dc.contributor.author","Jakobs, Stefan"],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Willig, Katrin I."],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2017-09-07T11:48:25Z"],["dc.date.available","2017-09-07T11:48:25Z"],["dc.date.issued","2012"],["dc.description.abstract","Lens-based fluorescence microscopy, which has long been limited in resolution to about 200 nanometers by diffraction, is rapidly evolving into a nanoscale imaging technique. Here, we show that the superresolution fluorescence microscopy called RESOLFT enables comparatively fast and continuous imaging of sensitive, nanosized features in living brain tissue. Using low-intensity illumination to switch photochromic fluorescent proteins reversibly between a fluorescent and a nonfluorescent state, we increased the resolution more than 3-fold over that of confocal microscopy in all dimensions. Dendritic spines located 10-50 mu m deep inside living organotypic hippocampal brain slices were recorded for hours without signs of degradation. Using a fast-switching protein increased the imaging speed 50-fold over reported RESOLFT schemes, which in turn enabled the recording of spontaneous and stimulated changes of dendritic actin filaments and spine morphology occurring on time scales from seconds to hours."],["dc.identifier.doi","10.1016/j.neuron.2012.07.028"],["dc.identifier.gro","3142464"],["dc.identifier.isi","000309198900011"],["dc.identifier.pmid","22998868"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/8574"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.publisher","Cell Press"],["dc.relation.issn","0896-6273"],["dc.title","Nanoscopy of Living Brain Slices with Low Light Levels"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.artnumber","540"],["dc.bibliographiccitation.issue","1"],["dc.bibliographiccitation.journal","Molecular Systems Biology"],["dc.bibliographiccitation.volume","7"],["dc.contributor.author","Kleine-Vehn, Jürgen"],["dc.contributor.author","Wabnik, Krzysztof"],["dc.contributor.author","Martinière, Alexandre"],["dc.contributor.author","Langowski, Lukasz"],["dc.contributor.author","Willig, Katrin"],["dc.contributor.author","Naramoto, Satoshi"],["dc.contributor.author","Leitner, Johannes"],["dc.contributor.author","Tanaka, Hirokazu"],["dc.contributor.author","Jakobs, Stefan"],["dc.contributor.author","Robert, Stéphanie"],["dc.contributor.author","Luschnig, Christian"],["dc.contributor.author","Govaerts, Willy"],["dc.contributor.author","Hell, Stefan W."],["dc.contributor.author","Runions, John"],["dc.contributor.author","Friml, Jiří"],["dc.date.accessioned","2017-09-07T11:43:23Z"],["dc.date.available","2017-09-07T11:43:23Z"],["dc.date.issued","2011"],["dc.description.abstract","Cell polarity reflected by asymmetric distribution of proteins at the plasma membrane is a fundamental feature of unicellular and multicellular organisms. It remains conceptually unclear how cell polarity is kept in cell wall-encapsulated plant cells. We have used super-resolution and semi-quantitative live-cell imaging in combination with pharmacological, genetic, and computational approaches to reveal insights into the mechanism of cell polarity maintenance in Arabidopsis thaliana. We show that polar-competent PIN transporters for the phytohormone auxin are delivered to the center of polar domains by super-polar recycling. Within the plasma membrane, PINs are recruited into non-mobile membrane clusters and their lateral diffusion is dramatically reduced, which ensures longer polar retention. At the circumventing edges of the polar domain, spatially defined internalization of escaped cargos occurs by clathrin-dependent endocytosis. Computer simulations confirm that the combination of these processes provides a robust mechanism for polarity maintenance in plant cells. Moreover, our study suggests that the regulation of lateral diffusion and spatially defined endocytosis, but not super-polar exocytosis have primary importance for PIN polarity maintenance. Molecular Systems Biology 7: 540; published online 25 October 2011; doi:10.1038/msb.2011.72"],["dc.identifier.doi","10.1038/msb.2011.72"],["dc.identifier.gro","3142654"],["dc.identifier.isi","000296652600004"],["dc.identifier.pmid","22027551"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/82"],["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","1744-4292"],["dc.title","Recycling, clustering, and endocytosis jointly maintain PIN auxin carrier polarity at the plasma membrane"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","158"],["dc.bibliographiccitation.issue","1"],["dc.bibliographiccitation.journal","Biophysical Journal"],["dc.bibliographiccitation.lastpage","163"],["dc.bibliographiccitation.volume","98"],["dc.contributor.author","Hein, Birka"],["dc.contributor.author","Willig, Katrin I."],["dc.contributor.author","Wurm, Christian A."],["dc.contributor.author","Westphal, Volker"],["dc.contributor.author","Jakobs, Stefan"],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2017-09-07T11:46:10Z"],["dc.date.available","2017-09-07T11:46:10Z"],["dc.date.issued","2010"],["dc.description.abstract","We show far-field fluorescence nanoscopy of different structural elements labeled with an organic dye within living mammalian cells. The diffraction barrier limiting far-field light microscopy is outperformed by using stimulated emission depletion. We used the tagging protein hAGT (SNAP-tag), which covalently binds benzy1guanine-substituted organic dyes, for labeling. Tetramethy1rhodamine was used to image the cytoskeleton (vimentin and microtubule-associated protein 2) as well as structures located at the cell membrane (caveolin and connexin-43) with a resolution down to 40 nm. Comparison with structures labeled with the yellow fluorescent protein Citrine validates this labeling approach. Nanoscopic movies showing the movement of connexin-43 clusters across the cell membrane evidence the capability of this technique to observe structural changes on the nanoscale over time. Pulsed or continuous-wave lasers for excitation and stimulated emission depletion yield images of similar resolution in living cells. Hence fusion proteins that bind modified organic dyes expand widely the application range of far-field fluorescence nanoscopy of living cells."],["dc.identifier.doi","10.1016/j.bpj.2009.09.053"],["dc.identifier.gro","3142983"],["dc.identifier.isi","000273433800018"],["dc.identifier.pmid","20074516"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/447"],["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","Stimulated Emission Depletion Nanoscopy of Living Cells Using SNAP-Tag Fusion Proteins"],["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.lastpage","579"],["dc.contributor.author","Hell, Stefan W."],["dc.contributor.author","Willig, Katrin I."],["dc.contributor.author","Dyba, Marcus"],["dc.contributor.author","Jakobs, Stefan"],["dc.contributor.author","Kastrup, Lars"],["dc.contributor.author","Westphal, Volker"],["dc.contributor.editor","Pawley, J. B."],["dc.date.accessioned","2017-09-07T11:53:05Z"],["dc.date.available","2017-09-07T11:53:05Z"],["dc.date.issued","2010"],["dc.identifier.doi","10.1007/978-0-387-45524-2_31"],["dc.identifier.gro","3145032"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/2723"],["dc.language.iso","en"],["dc.notes.intern","Crossref Import"],["dc.notes.status","final"],["dc.publisher","Springer Nature"],["dc.publisher.place","New York"],["dc.relation.isbn","978-0-387-25921-5"],["dc.relation.ispartof","Handbook of biological confocal microscopy"],["dc.title","Nanoscale Resolution with Focused Light: Stimulated Emission Depletion and Other Reversible Saturable Optical Fluorescence Transitions Microscopy Concepts"],["dc.type","book_chapter"],["dc.type.internalPublication","unknown"],["dc.type.peerReviewed","no"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","3970"],["dc.bibliographiccitation.issue","9"],["dc.bibliographiccitation.journal","Nano Letters"],["dc.bibliographiccitation.lastpage","3973"],["dc.bibliographiccitation.volume","11"],["dc.contributor.author","Willig, Katrin I."],["dc.contributor.author","Stiel, Andre C."],["dc.contributor.author","Brakemann, Tanja"],["dc.contributor.author","Jakobs, Stefan"],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2017-09-07T11:43:25Z"],["dc.date.available","2017-09-07T11:43:25Z"],["dc.date.issued","2011"],["dc.description.abstract","We demonstrate live-cell STED microscopy of two protein species using photochromic green fluorescent proteins as markers. The reversible photoswitching of two markers is implemented so that they can be discerned with a single excitation and STED wavelength and a single detection channel. Dual-label STED microscopy is shown in living mammalian cells."],["dc.identifier.doi","10.1021/nl202290w"],["dc.identifier.gro","3142671"],["dc.identifier.isi","000294790200079"],["dc.identifier.pmid","21786833"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/100"],["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","1530-6984"],["dc.title","Dual-Label STED Nanoscopy of Living Cells Using Photochromism"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.artnumber","443001"],["dc.bibliographiccitation.issue","44"],["dc.bibliographiccitation.journal","Journal of Physics D: Applied Physics"],["dc.bibliographiccitation.volume","48"],["dc.contributor.author","Hell, Stefan"],["dc.contributor.author","Sahl, Steffen J."],["dc.contributor.author","Bates, Mark"],["dc.contributor.author","Zhuang, Xiaowei"],["dc.contributor.author","Heintzmann, Rainer"],["dc.contributor.author","Booth, Martin J."],["dc.contributor.author","Bewersdorf, Jörg"],["dc.contributor.author","Shtengel, Gleb"],["dc.contributor.author","Hess, Harald"],["dc.contributor.author","Tinnefeld, Philip"],["dc.contributor.author","Honigmann, Alf"],["dc.contributor.author","Jakobs, Stefan"],["dc.contributor.author","Testa, Ilaria"],["dc.contributor.author","Cognet, Laurent"],["dc.contributor.author","Lounis, Brahim"],["dc.contributor.author","Ewers, Helge"],["dc.contributor.author","Davis, Simon J."],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Klenerman, David"],["dc.contributor.author","Willig, Katrin I."],["dc.contributor.author","Vicidomini, Giuseppe"],["dc.contributor.author","Castello, Marco"],["dc.contributor.author","Diaspro, Alberto"],["dc.contributor.author","Cordes, Thorben"],["dc.date.accessioned","2017-09-07T11:54:53Z"],["dc.date.available","2017-09-07T11:54:53Z"],["dc.date.issued","2015"],["dc.description.abstract","Far-field optical microscopy using focused light is an important tool in a number of scientific disciplines including chemical, (bio) physical and biomedical research, particularly with respect to the study of living cells and organisms. Unfortunately, the applicability of the optical microscope is limited, since the diffraction of light imposes limitations on the spatial resolution of the image. Consequently the details of, for example, cellular protein distributions, can be visualized only to a certain extent. Fortunately, recent years have witnessed the development of 'super-resolution' farfield optical microscopy (nanoscopy) techniques such as stimulated emission depletion (STED), ground state depletion (GSD), reversible saturated optical (fluorescence) transitions (RESOLFT), photoactivation localization microscopy (PALM), stochastic optical reconstruction microscopy (STORM), structured illumination microscopy (SIM) or saturated structured illumination microscopy (SSIM), all in one way or another addressing the problem of the limited spatial resolution of far-field optical microscopy. While SIM achieves a two-fold improvement in spatial resolution compared to conventional optical microscopy, STED, RESOLFT, PALM/STORM, or SSIM have all gone beyond, pushing the limits of optical image resolution to the nanometer scale. Consequently, all super-resolution techniques open new avenues of biomedical research. Because the field is so young, the potential capabilities of different super-resolution microscopy approaches have yet to be fully explored, and uncertainties remain when considering the best choice of methodology. Thus, even for experts, the road to the future is sometimes shrouded in mist. The super-resolution optical microscopy roadmap of Journal of Physics D: Applied Physics addresses this need for clarity. It provides guidance to the outstanding questions through a collection of short review articles from experts in the field, giving a thorough discussion on the concepts underlying super-resolution optical microscopy, the potential of different approaches, the importance of label optimization (such as reversible photoswitchable proteins) and applications in which these methods will have a significant impact. Mark Bates, Christian Eggeling"],["dc.identifier.doi","10.1088/0022-3727/48/44/443001"],["dc.identifier.gro","3141789"],["dc.identifier.isi","000365925800002"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/1090"],["dc.notes.intern","WoS Import 2017-03-10 / Funder: Medical Research Council [MC_UU_12010/9]"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.publisher","Iop Publishing Ltd"],["dc.relation.eissn","1361-6463"],["dc.relation.issn","0022-3727"],["dc.title","The 2015 super-resolution microscopy roadmap"],["dc.type","review"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","721"],["dc.bibliographiccitation.issue","9"],["dc.bibliographiccitation.journal","Nature Methods"],["dc.bibliographiccitation.lastpage","723"],["dc.bibliographiccitation.volume","3"],["dc.contributor.author","Willig, Katrin I."],["dc.contributor.author","Kellner, Robert R."],["dc.contributor.author","Medda, Rebecca"],["dc.contributor.author","Hein, Birka"],["dc.contributor.author","Jakobs, Stefan"],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2017-09-07T11:52:34Z"],["dc.date.available","2017-09-07T11:52:34Z"],["dc.date.issued","2006"],["dc.description.abstract","We report attainment of subdiffraction resolution using stimulated emission depletion (STED) microscopy with GFP-labeled samples. The similar to 70 nm lateral resolution attained in this study is demonstrated by imaging GFP-labeled viruses and the endoplasmic reticulum (ER) of a mammalian cell. Our results mark the advent of nanoscale biological microscopy with genetically encoded markers."],["dc.identifier.doi","10.1038/NMETH922"],["dc.identifier.gro","3143638"],["dc.identifier.isi","000240290300016"],["dc.identifier.pmid","16896340"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/1174"],["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","Nanoscale resolution in GFP-based microscopy"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]