Gwosch, Klaus C.

[["dc.bibliographiccitation.firstpage","606"],["dc.bibliographiccitation.issue","6325"],["dc.bibliographiccitation.journal","Science"],["dc.bibliographiccitation.lastpage","612"],["dc.bibliographiccitation.volume","355"],["dc.contributor.author","Balzarotti, Francisco"],["dc.contributor.author","Eilers, Yvan"],["dc.contributor.author","Gwosch, Klaus C."],["dc.contributor.author","Gynnå, Arvid H."],["dc.contributor.author","Westphal, Volker"],["dc.contributor.author","Stefani, Fernando D."],["dc.contributor.author","Elf, Johan"],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2018-01-17T13:33:22Z"],["dc.date.available","2018-01-17T13:33:22Z"],["dc.date.issued","2017"],["dc.description.abstract","We introduce MINFLUX, a concept for localizing photon emitters in space. By probing the emitter with a local intensity minimum of excitation light, MINFLUX minimizes the fluorescence photons needed for high localization precision. In our experiments, 22 times fewer fluorescence photons are required as compared to popular centroid localization. In superresolution microscopy, MINFLUX attained ~1-nm precision, resolving molecules only 6 nanometers apart. MINFLUX tracking of single fluorescent proteins increased the temporal resolution and the number of localizations per trace by a factor of 100, as demonstrated with diffusing 30S ribosomal subunits in living Escherichia coli As conceptual limits have not been reached, we expect this localization modality to break new ground for observing the dynamics, distribution, and structure of macromolecules in living cells and beyond."],["dc.identifier.arxiv","1611.03401v1"],["dc.identifier.doi","10.1126/science.aak9913"],["dc.identifier.pmid","28008086"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/11719"],["dc.language.iso","en"],["dc.notes.status","final"],["dc.relation.eissn","1095-9203"],["dc.subject","Physics - Optics"],["dc.subject","Physics - Optics"],["dc.subject","Physics - Biological Physics"],["dc.title","Nanometer resolution imaging and tracking of fluorescent molecules with minimal photon fluxes"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","6117"],["dc.bibliographiccitation.issue","24"],["dc.bibliographiccitation.journal","Proceedings of the National Academy of Sciences"],["dc.bibliographiccitation.lastpage","6122"],["dc.bibliographiccitation.volume","115"],["dc.contributor.author","Eilers, Yvan"],["dc.contributor.author","Ta, Haisen"],["dc.contributor.author","Gwosch, Klaus C."],["dc.contributor.author","Balzarotti, Francisco"],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2022-03-01T11:46:24Z"],["dc.date.available","2022-03-01T11:46:24Z"],["dc.date.issued","2018"],["dc.description.abstract","Compared with localization schemes solely based on evaluating patterns of molecular emission, the recently introduced single-molecule localization concept called MINFLUX and the fluorescence nanoscopies derived from it require up to orders of magnitude fewer emissions to attain single-digit nanometer resolution. Here, we demonstrate that the lower number of required fluorescence photons enables MINFLUX to detect molecular movements of a few nanometers at a temporal sampling of well below 1 millisecond. Using fluorophores attached to thermally fluctuating DNA strands as model systems, we demonstrate that measurement times as short as 400 microseconds suffice to localize fluorescent molecules with ∼2-nm precision. Such performance is out of reach for popular camera-based localization by centroid calculation of emission diffraction patterns. Since theoretical limits have not been reached, our results show that emerging MINFLUX nanoscopy bears great potential for dissecting the motions of individual (macro)molecules at hitherto-unattained combinations of spatial and temporal resolution."],["dc.description.abstract","Compared with localization schemes solely based on evaluating patterns of molecular emission, the recently introduced single-molecule localization concept called MINFLUX and the fluorescence nanoscopies derived from it require up to orders of magnitude fewer emissions to attain single-digit nanometer resolution. Here, we demonstrate that the lower number of required fluorescence photons enables MINFLUX to detect molecular movements of a few nanometers at a temporal sampling of well below 1 millisecond. Using fluorophores attached to thermally fluctuating DNA strands as model systems, we demonstrate that measurement times as short as 400 microseconds suffice to localize fluorescent molecules with ∼2-nm precision. Such performance is out of reach for popular camera-based localization by centroid calculation of emission diffraction patterns. Since theoretical limits have not been reached, our results show that emerging MINFLUX nanoscopy bears great potential for dissecting the motions of individual (macro)molecules at hitherto-unattained combinations of spatial and temporal resolution."],["dc.identifier.doi","10.1073/pnas.1801672115"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/103658"],["dc.language.iso","en"],["dc.notes.intern","DOI-Import GROB-531"],["dc.relation.eissn","1091-6490"],["dc.relation.issn","0027-8424"],["dc.rights.uri","https://creativecommons.org/licenses/by-nc-nd/4.0/"],["dc.title","MINFLUX monitors rapid molecular jumps with superior spatiotemporal resolution"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","962"],["dc.bibliographiccitation.issue","5"],["dc.bibliographiccitation.journal","Proceedings of the National Academy of Sciences of the United States of America"],["dc.bibliographiccitation.lastpage","967"],["dc.bibliographiccitation.volume","115"],["dc.contributor.author","Gregor, Carola"],["dc.contributor.author","Gwosch, Klaus C."],["dc.contributor.author","Sahl, Steffen J."],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2018-01-17T13:26:27Z"],["dc.date.available","2018-01-17T13:26:27Z"],["dc.date.issued","2018"],["dc.description.abstract","Bioluminescence imaging of single cells is often complicated by the requirement of exogenous luciferins that can be poorly cell-permeable or produce high background signal. Bacterial bioluminescence is unique in that it uses reduced flavin mononucleotide as a luciferin, which is abundant in all cells, making this system purely genetically encodable by the lux operon. Unfortunately, the use of bacterial bioluminescence has been limited by its low brightness compared with other luciferases. Here, we report the generation of an improved lux operon named ilux with an approximately sevenfold increased brightness when expressed in Escherichia coli; ilux can be used to image single E. coli cells with enhanced spatiotemporal resolution over several days. In addition, since only metabolically active cells produce bioluminescent signal, we show that ilux can be used to observe the effect of different antibiotics on cell viability on the single-cell level."],["dc.identifier.doi","10.1073/pnas.1715946115"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/11712"],["dc.language.iso","en"],["dc.notes.status","final"],["dc.title","Strongly enhanced bacterial bioluminescence with theiluxoperon for single-cell imaging"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","217"],["dc.bibliographiccitation.issue","2"],["dc.bibliographiccitation.journal","Nature Methods"],["dc.bibliographiccitation.lastpage","224"],["dc.bibliographiccitation.volume","17"],["dc.contributor.author","Gwosch, Klaus C."],["dc.contributor.author","Pape, Jasmin K."],["dc.contributor.author","Balzarotti, Francisco"],["dc.contributor.author","Hoess, Philipp"],["dc.contributor.author","Ellenberg, Jan"],["dc.contributor.author","Ries, Jonas"],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2022-03-01T11:46:02Z"],["dc.date.available","2022-03-01T11:46:02Z"],["dc.date.issued","2020"],["dc.identifier.doi","10.1038/s41592-019-0688-0"],["dc.identifier.pii","688"],["dc.identifier.pmid","31932776"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/103535"],["dc.identifier.url","https://sfb1286.uni-goettingen.de/literature/publications/43"],["dc.language.iso","en"],["dc.notes.intern","DOI-Import GROB-531"],["dc.relation","SFB 1286: Quantitative Synaptologie"],["dc.relation","SFB 1286 | A07: Der Aufbau des synaptischen Cytoskeletts"],["dc.relation.eissn","1548-7105"],["dc.relation.issn","1548-7091"],["dc.relation.workinggroup","RG Hell"],["dc.rights.uri","http://www.springer.com/tdm"],["dc.title","MINFLUX nanoscopy delivers 3D multicolor nanometer resolution in cells"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dc.type.subtype","original_ja"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","26491"],["dc.bibliographiccitation.issue","52"],["dc.bibliographiccitation.journal","Proceedings of the National Academy of Sciences of the United States of America"],["dc.bibliographiccitation.lastpage","26496"],["dc.bibliographiccitation.volume","116"],["dc.contributor.author","Gregor, Carola"],["dc.contributor.author","Pape, Jasmin K."],["dc.contributor.author","Gwosch, Klaus C."],["dc.contributor.author","Gilat, Tanja"],["dc.contributor.author","Sahl, Steffen J."],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2020-04-03T13:38:03Z"],["dc.date.available","2020-04-03T13:38:03Z"],["dc.date.issued","2019-12-02"],["dc.description.abstract","Bioluminescence-based imaging of living cells has become an important tool in biological and medical research. However, many bioluminescence imaging applications are limited by the requirement of an externally provided luciferin substrate and the low bioluminescence signal which restricts the sensitivity and spatiotemporal resolution. The bacterial bioluminescence system is fully genetically encodable and hence produces autonomous bioluminescence without an external luciferin, but its brightness in cell types other than bacteria has, so far, not been sufficient for imaging single cells. We coexpressed codon-optimized forms of the bacterial luxCDABE and frp genes from multiple plasmids in different mammalian cell lines. Our approach produces high luminescence levels that are comparable to firefly luciferase, thus enabling autonomous bioluminescence microscopy of mammalian cells."],["dc.identifier.doi","10.1073/pnas.1913616116"],["dc.identifier.pmid","31792180"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/63635"],["dc.language.iso","en"],["dc.relation.eissn","1091-6490"],["dc.relation.issn","0027-8424"],["dc.relation.issn","1091-6490"],["dc.title","Autonomous bioluminescence imaging of single mammalian cells with the bacterial bioluminescence system"],["dc.type","journal_article"],["dc.type.internalPublication","no"],["dspace.entity.type","Publication"]]