Sahl, Steffen J.

[["dc.bibliographiccitation.artnumber","11354"],["dc.bibliographiccitation.journal","Scientific Reports"],["dc.bibliographiccitation.volume","7"],["dc.contributor.author","Oracz, Joanna"],["dc.contributor.author","Westphal, Volker"],["dc.contributor.author","Radzewicz, Czesław"],["dc.contributor.author","Sahl, Steffen J."],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2018-01-17T13:41:07Z"],["dc.date.available","2018-01-17T13:41:07Z"],["dc.date.issued","2017-09-12"],["dc.description.abstract","In STED (stimulated emission depletion) nanoscopy, the resolution and signal are limited by the fluorophore de-excitation efficiency and photobleaching. Here, we investigated their dependence on the pulse duration and power of the applied STED light for the popular 750 nm wavelength. In experiments with red- and orange-emitting dyes, the pulse duration was varied from the sub-picosecond range up to continuous-wave conditions, with average powers up to 200 mW at 80 MHz repetition rate, i.e. peak powers up to 1 kW and pulse energies up to 2.5 nJ. We demonstrate the dependence of bleaching on pulse duration, which dictates the optimal parameters of how to deliver the photons required for transient fluorophore silencing. Measurements with the dye ATTO647N reveal that the bleaching of excited molecules scales with peak power with a single effective order ~1.4. This motivates peak power reduction while maintaining the number of STED-light photons, in line with the superior resolution commonly achieved for nanosecond STED pulses. Other dyes (ATTO590, STAR580, STAR635P) exhibit two distinctive bleaching regimes for constant pulse energy, one with strong dependence on peak power, one nearly independent. We interpret the results within a photobleaching model that guides quantitative predictions of resolution and bleaching."],["dc.identifier.doi","10.1038/s41598-017-09902-x"],["dc.identifier.pmid","28900102"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/11728"],["dc.language.iso","en"],["dc.notes.status","final"],["dc.relation.eissn","2045-2322"],["dc.title","Photobleaching in STED nanoscopy and its dependence on the photon flux applied for reversible silencing of the fluorophore"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["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.firstpage","1072"],["dc.bibliographiccitation.issue","9"],["dc.bibliographiccitation.journal","Nature Methods"],["dc.bibliographiccitation.lastpage","1075"],["dc.bibliographiccitation.volume","19"],["dc.contributor.author","Ostersehlt, Lynn M."],["dc.contributor.author","Jans, Daniel C."],["dc.contributor.author","Wittek, Anna"],["dc.contributor.author","Keller-Findeisen, Jan"],["dc.contributor.author","Inamdar, Kaushik"],["dc.contributor.author","Sahl, Steffen J."],["dc.contributor.author","Hell, Stefan W."],["dc.contributor.author","Jakobs, Stefan"],["dc.date.accessioned","2022-10-04T10:21:07Z"],["dc.date.available","2022-10-04T10:21:07Z"],["dc.date.issued","2022"],["dc.description.abstract","Abstract\n MINimal fluorescence photon FLUXes (MINFLUX) nanoscopy, providing photon-efficient fluorophore localizations, has brought about three-dimensional resolution at nanometer scales. However, by using an intrinsic on–off switching process for single fluorophore separation, initial MINFLUX implementations have been limited to two color channels. Here we show that MINFLUX can be effectively combined with sequentially multiplexed DNA-based labeling (DNA-PAINT), expanding MINFLUX nanoscopy to multiple molecular targets. Our method is exemplified with three-color recordings of mitochondria in human cells."],["dc.identifier.doi","10.1038/s41592-022-01577-1"],["dc.identifier.pii","1577"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/114334"],["dc.language.iso","en"],["dc.notes.intern","DOI-Import GROB-600"],["dc.relation.eissn","1548-7105"],["dc.relation.issn","1548-7091"],["dc.rights.uri","https://creativecommons.org/licenses/by/4.0"],["dc.title","DNA-PAINT MINFLUX nanoscopy"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.artnumber","7127"],["dc.bibliographiccitation.journal","Nature Communications"],["dc.bibliographiccitation.volume","6"],["dc.contributor.author","Hanne, Janina"],["dc.contributor.author","Falk, Henning J."],["dc.contributor.author","Görlitz, Frederik"],["dc.contributor.author","Hoyer, Patrick"],["dc.contributor.author","Engelhardt, Johann"],["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 widely popular class of quantum-dot molecular labels could so far not be utilized as standard fluorescent probes in STED (stimulated emission depletion) nanoscopy. This is because broad quantum-dot excitation spectra extend deeply into the spectral bands used for STED, thus compromising the transient fluorescence silencing required for attaining super-resolution. Here we report the discovery that STED nanoscopy of several red-emitting commercially available quantum dots is in fact successfully realized by the increasingly popular 775nm STED laser light. A resolution of presently similar to 50 nm is demonstrated for single quantum dots, and sub-diffraction resolution is further shown for imaging of quantum-dot-labelled vimentin filaments in fibroblasts. The high quantum-dot photostability enables repeated STED recordings with 41,000 frames. In addition, we have evidence that the tendency of quantum-dot labels to blink is largely suppressed by combined action of excitation and STED beams. Quantum-dot STED significantly expands the realm of application of STED nanoscopy, and, given the high stability of these probes, holds promise for extended time-lapse imaging."],["dc.identifier.doi","10.1038/ncomms8127"],["dc.identifier.gro","3141908"],["dc.identifier.isi","000355533000015"],["dc.identifier.pmid","25980788"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/2411"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10 / Funder: Deutsche Forschungsgemeinschaft"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","2041-1723"],["dc.title","STED nanoscopy with fluorescent quantum dots"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.journal","Scientific Reports"],["dc.bibliographiccitation.volume","8"],["dc.contributor.author","Gregor, Carola"],["dc.contributor.author","Sidenstein, Sven C."],["dc.contributor.author","Andresen, Martin"],["dc.contributor.author","Sahl, Steffen J."],["dc.contributor.author","Danzl, Johann G."],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2018-04-23T11:48:22Z"],["dc.date.available","2018-04-23T11:48:22Z"],["dc.date.issued","2018"],["dc.description.abstract","The reversibly switchable fluorescent proteins (RSFPs) commonly used for RESOLFT nanoscopy have been developed from fluorescent proteins of the GFP superfamily. These proteins are bright, but exhibit several drawbacks such as relatively large size, oxygen-dependence, sensitivity to low pH, and limited switching speed. Therefore, RSFPs from other origins with improved properties need to be explored. Here, we report the development of two RSFPs based on the LOV domain of the photoreceptor protein YtvA from Bacillus subtilis. LOV domains obtain their fluorescence by association with the abundant cellular cofactor flavin mononucleotide (FMN). Under illumination with blue and ultraviolet light, they undergo a photocycle, making these proteins inherently photoswitchable. Our first improved variant, rsLOV1, can be used for RESOLFT imaging, whereas rsLOV2 proved useful for STED nanoscopy of living cells with a resolution of down to 50 nm. In addition to their smaller size compared to GFP-related proteins (17 kDa instead of 27 kDa) and their usability at low pH, rsLOV1 and rsLOV2 exhibit faster switching kinetics, switching on and off 3 times faster than rsEGFP2, the fastest-switching RSFP reported to date. Therefore, LOV-domain-based RSFPs have potential for applications where the switching speed of GFP-based proteins is limiting."],["dc.identifier.doi","10.1038/s41598-018-19947-1"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/13496"],["dc.language.iso","en"],["dc.notes.status","zu prüfen"],["dc.relation.issn","2045-2322"],["dc.title","Novel reversibly switchable fluorescent proteins for RESOLFT and STED nanoscopy engineered from the bacterial photoreceptor YtvA"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.artnumber","jmi.13143"],["dc.bibliographiccitation.journal","Journal of Microscopy"],["dc.contributor.author","Engelhardt, Johann"],["dc.contributor.author","Ellerhoff, Beatrice"],["dc.contributor.author","Gürth, Clara‐Marie"],["dc.contributor.author","Sahl, Steffen J."],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2022-10-04T10:21:47Z"],["dc.date.available","2022-10-04T10:21:47Z"],["dc.date.issued","2022"],["dc.identifier.doi","10.1111/jmi.13143"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/114501"],["dc.language.iso","en"],["dc.notes.intern","DOI-Import GROB-600"],["dc.relation.eissn","1365-2818"],["dc.relation.issn","0022-2720"],["dc.title","Designing chromatic optical retarder stacks for segmented next‐generation easySTED phase plates"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","827"],["dc.bibliographiccitation.issue","9"],["dc.bibliographiccitation.journal","Nature Methods"],["dc.bibliographiccitation.lastpage","830"],["dc.bibliographiccitation.volume","12"],["dc.contributor.author","Schneider, Jale"],["dc.contributor.author","Zahn, Jasmin"],["dc.contributor.author","Maglione, Marta"],["dc.contributor.author","Sigrist, Stephan J."],["dc.contributor.author","Marquard, Jonas"],["dc.contributor.author","Chojnacki, Jakub"],["dc.contributor.author","Kraeusslich, Hans-Georg"],["dc.contributor.author","Sahl, Steffen J."],["dc.contributor.author","Engelhardt, Johann"],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2017-09-07T11:43:35Z"],["dc.date.available","2017-09-07T11:43:35Z"],["dc.date.issued","2015"],["dc.description.abstract","Electro-optical scanning (>1,000 frames/s) with pixel dwell times on the order of the lifetime of the fluorescent molecular state renders stimulated emission depletion (STED) nanoscopy temporally stochastic. Photon detection from a molecule occurs stochastically in one of several scanning frames, and the spatial origin of the photon is known with subdiffraction precision. Images are built up by binning consecutive frames, making the time resolution freely adjustable. We demonstrated nanoscopy of vesicle motions in living Drosophila larvae and the cellular uptake of viral particles with 5- to 10-ms temporal resolution."],["dc.identifier.doi","10.1038/NMETH.3481"],["dc.identifier.gro","3141839"],["dc.identifier.isi","000360586700025"],["dc.identifier.pmid","26214129"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/1645"],["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","1548-7105"],["dc.relation.issn","1548-7091"],["dc.title","Ultrafast, temporally stochastic STED nanoscopy of millisecond dynamics"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.artnumber","527a"],["dc.bibliographiccitation.issue","6285"],["dc.bibliographiccitation.journal","Science"],["dc.bibliographiccitation.volume","352"],["dc.contributor.author","Sahl, Steffen J."],["dc.contributor.author","Balzarotti, Francisco"],["dc.contributor.author","Keller-Findeisen, Jan"],["dc.contributor.author","Leutenegger, Marcel"],["dc.contributor.author","Westphal, Volker"],["dc.contributor.author","Egner, Alexander"],["dc.contributor.author","Lavoie-Cardinal, Flavie"],["dc.contributor.author","Chmyrov, Andriy"],["dc.contributor.author","Grotjohann, Tim"],["dc.contributor.author","Jakobs, Stefan"],["dc.date.accessioned","2017-09-07T11:54:33Z"],["dc.date.available","2017-09-07T11:54:33Z"],["dc.date.issued","2016"],["dc.description.abstract","Li et al. (Research Articles, 28 August 2015, aab3500) purport to present solutions to longstanding challenges in live-cell microscopy, reporting relatively fast acquisition times in conjunction with improved image resolution. We question the methods' reliability to visualize specimen features at sub-100-nanometer scales, because the mandatory mathematical processing of the recorded data leads to artifacts that are either difficult or impossible to disentangle from real features. We are also concerned about the chosen approach of subjectively comparing images from different super-resolution methods, as opposed to using quantitative measures."],["dc.identifier.doi","10.1126/science.aad7983"],["dc.identifier.gro","3141696"],["dc.identifier.isi","000374998600028"],["dc.identifier.pmid","27126030"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/58"],["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","1095-9203"],["dc.relation.issn","0036-8075"],["dc.title","Comment on \"Extended-resolution structured illumination imaging of endocytic and cytoskeletal dynamics\""],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","letter_note"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","3442"],["dc.bibliographiccitation.issue","13"],["dc.bibliographiccitation.journal","Proceedings of the National Academy of Sciences"],["dc.bibliographiccitation.lastpage","3446"],["dc.bibliographiccitation.volume","113"],["dc.contributor.author","Hoyer, Patrick"],["dc.contributor.author","Medeiros, Gustavo de"],["dc.contributor.author","Balazs, Balint"],["dc.contributor.author","Norlin, Nils"],["dc.contributor.author","Besir, Christina"],["dc.contributor.author","Hanne, Janina"],["dc.contributor.author","Kraeusslich, Hans-Georg"],["dc.contributor.author","Engelhardt, Johann"],["dc.contributor.author","Sahl, Steffen J."],["dc.contributor.author","Hell, Stefan"],["dc.contributor.author","Hufnagel, Lars"],["dc.date.accessioned","2017-09-07T11:54:34Z"],["dc.date.available","2017-09-07T11:54:34Z"],["dc.date.issued","2016"],["dc.description.abstract","We present a plane-scanning RESOLFT [reversible saturable/switchable optical (fluorescence) transitions] light-sheet (LS) nanoscope, which fundamentally overcomes the diffraction barrier in the axial direction via confinement of the fluorescent molecular state to a sheet of subdiffraction thickness around the focal plane. To this end, reversibly switchable fluorophores located right above and below the focal plane are transferred to a nonfluorescent state at each scanning step. LS-RESOLFT nanoscopy offers wide-field 3D imaging of living biological specimens with low light dose and axial resolution far beyond the diffraction barrier. We demonstrate optical sections that are thinner by 5-12-fold compared with their conventional diffraction-limited LS analogs."],["dc.identifier.doi","10.1073/pnas.1522292113"],["dc.identifier.gro","3141706"],["dc.identifier.isi","000372876400034"],["dc.identifier.pmid","26984498"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/169"],["dc.notes.intern","WoS Import 2017-03-10 / Funder: Federal Ministry of Education and Research [FKZ:13N11173]"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","PUB_WoS_Import"],["dc.publisher","Natl Acad Sciences"],["dc.relation.issn","0027-8424"],["dc.title","Breaking the diffraction limit of light-sheet fluorescence microscopy by RESOLFT"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","E8047"],["dc.bibliographiccitation.issue","34"],["dc.bibliographiccitation.journal","Proceedings of the National Academy of Sciences of the United States of America"],["dc.bibliographiccitation.lastpage","E8056"],["dc.bibliographiccitation.volume","115"],["dc.contributor.author","Masch, Jennifer-Magdalena"],["dc.contributor.author","Steffens, Heinz"],["dc.contributor.author","Fischer, Joachim"],["dc.contributor.author","Engelhardt, Johann"],["dc.contributor.author","Hubrich, Jasmine"],["dc.contributor.author","Keller-Findeisen, Jan"],["dc.contributor.author","D'Este, Elisa"],["dc.contributor.author","Urban, Nicolai T."],["dc.contributor.author","Grant, Seth G. N."],["dc.contributor.author","Sahl, Steffen J."],["dc.contributor.author","Kamin, Dirk"],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2018-11-16T10:48:20Z"],["dc.date.accessioned","2021-10-27T13:21:10Z"],["dc.date.available","2018-11-16T10:48:20Z"],["dc.date.available","2021-10-27T13:21:10Z"],["dc.date.issued","2018"],["dc.identifier.doi","10.1073/pnas.1807104115"],["dc.identifier.pmid","30082388"],["dc.identifier.purl","https://resolver.sub.uni-goettingen.de/purl?gs-1/15631"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/91999"],["dc.identifier.url","https://sfb1286.uni-goettingen.de/literature/publications/37"],["dc.language.iso","en"],["dc.notes.intern","Migrated from goescholar"],["dc.relation","SFB 1286: Quantitative Synaptologie"],["dc.relation","SFB 1286 | A07: Der Aufbau des synaptischen Cytoskeletts"],["dc.relation.orgunit","Universitätsmedizin Göttingen"],["dc.relation.workinggroup","RG D’Este"],["dc.relation.workinggroup","RG Hell"],["dc.rights.uri","https://goescholar.uni-goettingen.de/licenses"],["dc.title","Robust nanoscopy of a synaptic protein in living mice by organic-fluorophore labeling"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.subtype","original_ja"],["dc.type.version","published_version"],["dspace.entity.type","Publication"]]