Gregor, Carola

[["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","46492"],["dc.bibliographiccitation.journal","Scientific Reports"],["dc.bibliographiccitation.volume","7"],["dc.contributor.author","Winter, Franziska R."],["dc.contributor.author","Loidolt, Maria"],["dc.contributor.author","Westphal, Volker"],["dc.contributor.author","Butkevich, Alexey N."],["dc.contributor.author","Gregor, Carola"],["dc.contributor.author","Sahl, Steffen J."],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2018-01-17T13:38:46Z"],["dc.date.available","2018-01-17T13:38:46Z"],["dc.date.issued","2017-04-18"],["dc.description.abstract","The extension of fluorescence nanoscopy to larger numbers of molecular species concurrently visualized by distinct markers is of great importance for advanced biological applications. To date, up to four markers had been distinguished in STED experiments featuring comparatively elaborate imaging schemes and optical setups, and exploiting various properties of the fluorophores. Here we present a simple yet versatile STED design for multicolour imaging below the diffraction limit. A hyperspectral detection arrangement (hyperSTED) collects the fluorescence in four spectral channels, allowing the separation of four markers with only one excitation wavelength and a single STED beam. Unmixing of the different marker signals based on the simultaneous readout of all channels is performed with a non-negative matrix factorization algorithm. We illustrate the approach showing four-colour nanoscopy of fixed and living cellular samples."],["dc.identifier.doi","10.1038/srep46492"],["dc.identifier.pmid","28417977"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/11725"],["dc.language.iso","en"],["dc.notes.status","final"],["dc.relation.eissn","2045-2322"],["dc.title","Multicolour nanoscopy of fixed and living cells with a single STED beam and hyperspectral detection"],["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.artnumber","577"],["dc.bibliographiccitation.firstpage","1"],["dc.bibliographiccitation.journal","Nature communications"],["dc.bibliographiccitation.lastpage","9"],["dc.bibliographiccitation.volume","8"],["dc.contributor.author","Richardson, Douglas S."],["dc.contributor.author","Gregor, Carola"],["dc.contributor.author","Winter, Franziska R."],["dc.contributor.author","Urban, Nicolai T."],["dc.contributor.author","Sahl, Steffen J."],["dc.contributor.author","Willig, Katrin I."],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2018-01-17T13:31:10Z"],["dc.date.available","2018-01-17T13:31:10Z"],["dc.date.issued","2017"],["dc.description.abstract","Fluorescence-based biosensors have become essential tools for modern biology, allowing real-time monitoring of biological processes within living cells. Intracellular fluorescent pH probes comprise one of the most widely used families of biosensors in microscopy. One key application of pH probes has been to monitor the acidification of vesicles during endocytosis, an essential function that aids in cargo sorting and degradation. Prior to the development of super-resolution fluorescence microscopy (nanoscopy), investigation of endosomal dynamics in live cells remained difficult as these structures lie at or below the ~250 nm diffraction limit of light microscopy. Therefore, to aid in investigations of pH dynamics during endocytosis at the nanoscale, we have specifically designed a family of ratiometric endosomal pH probes for use in live-cell STED nanoscopy.Ratiometric fluorescent pH probes are useful tools to monitor acidification of vesicles during endocytosis, but the size of vesicles is below the diffraction limit. Here the authors develop a family of ratiometric pH sensors for use in STED super-resolution microscopy, and optimize their delivery to endosomes."],["dc.identifier.doi","10.1038/s41467-017-00606-4"],["dc.identifier.pmid","28924139"],["dc.identifier.purl","https://resolver.sub.uni-goettingen.de/purl?gs-1/16496"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/11717"],["dc.language.iso","en"],["dc.notes.intern","Merged from goescholar"],["dc.notes.status","final"],["dc.relation.eissn","2041-1723"],["dc.rights","CC BY 4.0"],["dc.rights.uri","https://creativecommons.org/licenses/by/4.0"],["dc.title","SRpHi ratiometric pH biosensors for super-resolution microscopy"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dc.type.version","published_version"],["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"]]