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.firstpage","L01"],["dc.bibliographiccitation.issue","1"],["dc.bibliographiccitation.journal","Biophysical Journal"],["dc.bibliographiccitation.lastpage","L03"],["dc.bibliographiccitation.volume","106"],["dc.contributor.author","Willig, Katrin I."],["dc.contributor.author","Steffens, Heinz"],["dc.contributor.author","Gregor, Carola"],["dc.contributor.author","Herholt, Alexander"],["dc.contributor.author","Rossner, Moritz J."],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2017-09-07T11:46:54Z"],["dc.date.available","2017-09-07T11:46:54Z"],["dc.date.issued","2014"],["dc.description.abstract","We demonstrate superresolution fluorescence microscopy (nanoscopy) of protein distributions in a mammalian brain in vivo. Stimulated emission depletion microscopy reveals the morphology of the filamentous actin in dendritic spines down to 40 mu m in the molecular layer of the visual cortex of an anesthetized mouse. Consecutive recordings at 43-70 nm resolution reveal dynamical changes in spine morphology."],["dc.identifier.doi","10.1016/j.bpj.2013.11.1119"],["dc.identifier.gro","3142199"],["dc.identifier.isi","000329407700001"],["dc.identifier.pmid","24411266"],["dc.identifier.purl","https://resolver.sub.uni-goettingen.de/purl?gs-1/11365"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/5632"],["dc.language.iso","en"],["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.relation.eissn","1542-0086"],["dc.relation.issn","0006-3495"],["dc.rights","Goescholar"],["dc.rights.uri","https://goescholar.uni-goettingen.de/licenses"],["dc.title","Nanoscopy of Filamentous Actin in Cortical Dendrites of a Living Mouse"],["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","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","122"],["dc.bibliographiccitation.issue","2"],["dc.bibliographiccitation.journal","Nature Photonics"],["dc.bibliographiccitation.lastpage","128"],["dc.bibliographiccitation.volume","10"],["dc.contributor.author","Danzl, Johann G."],["dc.contributor.author","Sidenstein, Sven C."],["dc.contributor.author","Gregor, Carola"],["dc.contributor.author","Urban, Nicolai T."],["dc.contributor.author","Ilgen, Peter"],["dc.contributor.author","Jakobs, Stefan"],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2017-09-07T11:54:41Z"],["dc.date.available","2017-09-07T11:54:41Z"],["dc.date.issued","2016"],["dc.description.abstract","Far-field super-resolution fluorescence microscopy discerns fluorophores residing closer than the diffraction barrier by briefly transferring them in different (typically ON and OFF) states before detection. In coordinate-targeted super-resolution variants, such as stimulated emission depletion (STED) microscopy, this state difference is created by the intensity minima and maxima of an optical pattern, causing all fluorophores to assume the off state, for instance, except at the minima. Although strong spatial confinement of the on state enables high resolution, it also subjects the fluorophores to excess intensities and state cycles at the maxima. Here, we address these issues by driving the fluorophores into a second off state that is inert to the excess light. By using reversibly switchable fluorescent proteins as labels, our approach reduces bleaching and enhances resolution and contrast in live-cell STED microscopy. Using two or more transitions to off states is a useful strategy for augmenting the power of coordinate-targeted super-resolution microscopy."],["dc.identifier.doi","10.1038/NPHOTON.2015.266"],["dc.identifier.gro","3141737"],["dc.identifier.isi","000369321400015"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/513"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10 / Funder: European Union [PIEF-GA-2011-299283]; Korber Foundation"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.eissn","1749-4893"],["dc.relation.issn","1749-4885"],["dc.title","Coordinate-targeted fluorescence nanoscopy with multiple off states"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["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","186a"],["dc.bibliographiccitation.issue","3"],["dc.bibliographiccitation.journal","Biophysical Journal"],["dc.bibliographiccitation.lastpage","187a"],["dc.bibliographiccitation.volume","112"],["dc.contributor.author","Danzl, Johann G."],["dc.contributor.author","Sidenstein, Sven"],["dc.contributor.author","Gregor, Carola"],["dc.contributor.author","Urban, Nicolai"],["dc.contributor.author","Ilgen, Peter"],["dc.contributor.author","Jakobs, Stefan"],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2022-03-01T11:44:58Z"],["dc.date.available","2022-03-01T11:44:58Z"],["dc.date.issued","2017"],["dc.identifier.doi","10.1016/j.bpj.2016.11.1034"],["dc.identifier.pii","S0006349516320641"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/103178"],["dc.language.iso","en"],["dc.notes.intern","DOI-Import GROB-531"],["dc.relation.issn","0006-3495"],["dc.title","Coordinate-Targeted Fluorescence Nanoscopy with Multiple Off-States"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.journal","Acta Physiologica / Supplement"],["dc.bibliographiccitation.volume","213"],["dc.contributor.author","Hell, Stefan"],["dc.contributor.author","Willig, Katrin I."],["dc.contributor.author","Steffens, Heinz"],["dc.contributor.author","Wegner, W."],["dc.contributor.author","Gregor, Carola"],["dc.date.accessioned","2017-09-07T11:45:51Z"],["dc.date.available","2017-09-07T11:45:51Z"],["dc.date.issued","2015"],["dc.format.extent","18"],["dc.identifier.doi","10.1111/apha.12481"],["dc.identifier.gro","3145538"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/3247"],["dc.notes.intern","lifescience"],["dc.notes.status","public"],["dc.notes.submitter","oschaef1"],["dc.publisher","Wiley-Blackwell"],["dc.relation.conference","94th Annual Meeting of the German Physiological Society"],["dc.relation.eissn","1748-1716"],["dc.relation.eventend","2015-03-07"],["dc.relation.eventlocation","Magdeburg"],["dc.relation.eventstart","2015-03-05"],["dc.relation.issn","1748-1708"],["dc.title","In vivo STED microscopy of the visual cortex of adult mice"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dc.type.peerReviewed","no"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","2497"],["dc.bibliographiccitation.issue","8"],["dc.bibliographiccitation.journal","The Journal of Physiology"],["dc.bibliographiccitation.lastpage","2517"],["dc.bibliographiccitation.volume","595"],["dc.contributor.author","Bader, Almke"],["dc.contributor.author","Bintig, Willem"],["dc.contributor.author","Begandt, Daniela"],["dc.contributor.author","Klett, Anne"],["dc.contributor.author","Siller, Ina G."],["dc.contributor.author","Gregor, Carola"],["dc.contributor.author","Schaarschmidt, Frank"],["dc.contributor.author","Weksler, Babette"],["dc.contributor.author","Romero, Ignacio"],["dc.contributor.author","Couraud, Pierre-Olivier"],["dc.contributor.author","Ngezahayo, Anaclet"],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2022-03-01T11:47:11Z"],["dc.date.available","2022-03-01T11:47:11Z"],["dc.date.issued","2017"],["dc.description.abstract","The human cerebral microvascular endothelial cell line hCMEC/D3 was used to characterize the physiological link between adenosine receptors and the gap junction coupling in endothelial cells of the blood–brain barrier. Expressed adenosine receptor subtypes and connexin (Cx) isoforms were identified by RT‐PCR. Scrape loading/dye transfer was used to evaluate the impact of the A2A and A2B adenosine receptor subtype agonist 2‐phenylaminoadenosine (2‐PAA) on the gap junction coupling. We found that 2‐PAA stimulated cAMP synthesis and enhanced gap junction coupling in a concentration‐dependent manner. This enhancement was accompanied by an increase in gap junction plaques formed by Cx43. Inhibition of protein kinase A did not affect the 2‐PAA‐related enhancement of gap junction coupling. In contrast, the cyclic nucleotide‐gated (CNG) channel inhibitor l‐cis‐diltiazem, as well as the chelation of intracellular Ca2+ with BAPTA, or the absence of external Ca2+, suppressed the 2‐PAA‐related enhancement of gap junction coupling. Moreover, we observed a 2‐PAA‐dependent activation of CNG channels by a combination of electrophysiology and pharmacology. In conclusion, the stimulation of adenosine receptors in hCMEC/D3 cells induces a Ca2+ influx by opening CNG channels in a cAMP‐dependent manner. Ca2+ in turn induces the formation of new gap junction plaques and a consecutive sustained enhancement of gap junction coupling. The report identifies CNG channels as a physiological link that integrates gap junction coupling into the adenosine receptor‐dependent signalling of endothelial cells of the blood–brain barrier."],["dc.identifier.doi","10.1113/JP273150"],["dc.identifier.doi","10.1113/jp273150"],["dc.identifier.gro","3142362"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/103943"],["dc.language.iso","en"],["dc.notes.intern","DOI-Import GROB-531"],["dc.notes.status","final"],["dc.relation.issn","0022-3751"],["dc.title","Adenosine receptors regulate gap junction coupling of the human cerebral microvascular endothelial cells hCMEC/D3 by Ca 2+ influx through cyclic nucleotide-gated channels"],["dc.title.alternative","CNG channels as link between adenosine receptors and gap junctions"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dc.type.peerReviewed","no"],["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"]]