Wurm, Christian Andreas

[["dc.bibliographiccitation.firstpage","337"],["dc.bibliographiccitation.issue","2"],["dc.bibliographiccitation.journal","European Journal of Organic Chemistry"],["dc.bibliographiccitation.lastpage","349"],["dc.bibliographiccitation.volume","2015"],["dc.contributor.author","Mitronova, Gyuzel Yu"],["dc.contributor.author","Polyakova, Svetlana"],["dc.contributor.author","Wurm, Christian Andreas"],["dc.contributor.author","Kolmakov, Kirill"],["dc.contributor.author","Wolfram, Thomas"],["dc.contributor.author","Meineke, Dirk N. H."],["dc.contributor.author","Belov, Vladimir N."],["dc.contributor.author","John, Michael"],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2017-09-07T11:44:46Z"],["dc.date.available","2017-09-07T11:44:46Z"],["dc.date.issued","2015"],["dc.description.abstract","Aromatic nucleophilic substitution (SNAr) of fluorine in 9-(3'-carboxy-4',5',6',7'-tetrafluorophenyl) groups of xanthene dyes constitutes a powerful tool in dye design. Thiols and amines regioselectively replace F-6'. This approach enables additional hydrophilic residues or functional groups required for bioconjugation to be introduced. By using this methodology, a \"bright\" and photostable dye for two-color superresolution microscopy was synthesized (with absorption and emission maxima at 604 and 627 nm, respectively). In the case of red-emitting rhodamine dyes with 3'-carboxy-4',5',7'-trifluorophenyl residues, two-dimensional NMR techniques and a chemical transformation were used to prove the precise position of the additional substituent - a carboxylic acid group linked through the S-atom at C-6'. Furthermore, simple H-1 NMR spectra reliably permit the position of the additional carboxy substituent in the 3'-carboxyphenyl ring (at C-5' or C-6') to be established. Information on the exact position of this substituent is significant for the design of molecular probes and for the prediction of the properties of their bioconjugates."],["dc.identifier.doi","10.1002/ejoc.201403269"],["dc.identifier.gro","3141988"],["dc.identifier.isi","000347722900011"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/3301"],["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","1099-0690"],["dc.relation.issn","1434-193X"],["dc.title","Functionalization of the meso-Phenyl Ring of Rhodamine Dyes Through SNAr with Sulfur Nucleophiles: Synthesis, Biophysical Characterizations, and Comprehensive NMR Analysis"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","522"],["dc.bibliographiccitation.issue","3"],["dc.bibliographiccitation.journal","Photochemical & Photobiological Sciences"],["dc.bibliographiccitation.lastpage","532"],["dc.bibliographiccitation.volume","11"],["dc.contributor.author","Kolmakov, Kirill"],["dc.contributor.author","Wurm, Christian Andreas"],["dc.contributor.author","Sednev, Maksim V."],["dc.contributor.author","Bossi, Mariano L."],["dc.contributor.author","Belov, Vladimir N."],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2017-09-07T11:43:09Z"],["dc.date.available","2017-09-07T11:43:09Z"],["dc.date.issued","2012"],["dc.description.abstract","Caged near-IR emitting fluorescent dyes are in high demand in optical microscopy but up to now were unavailable. We discovered that the combination of a carbopyronine dye core and a photosensitive 2-diazo-1-indanone residue leads to masked near-IR emitting fluorescent dyes. Illumination of these caged dyes with either UV or visible light (lambda < 420 nm) efficiently generates fluorescent compounds with absorption and emission at 635 nm and 660 nm, respectively. A high-yielding synthetic route with attractive possibilities for further dye design is described in detail. Good photostability, high contrast, and a large fluorescence quantum yield after uncaging are the most important features of the new compounds for non-invasive imaging in high-resolution optical microscopy. For use in immunolabelling the caged dyes were decorated with a (hydrophilic) linker and an (activated) carboxyl group."],["dc.identifier.doi","10.1039/c1pp05321c"],["dc.identifier.gro","3142604"],["dc.identifier.isi","000300991600012"],["dc.identifier.pmid","22218703"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/27"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10 / Funder: Bundesministerium fur Bildung und Forschung (BMBF) [513, FKZ 13N11066]"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.eissn","1474-9092"],["dc.relation.issn","1474-905X"],["dc.title","Masked red-emitting carbopyronine dyes with photosensitive 2-diazo-1-indanone caging group"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","3598"],["dc.bibliographiccitation.issue","20"],["dc.bibliographiccitation.journal","Angewandte Chemie"],["dc.bibliographiccitation.lastpage","3602"],["dc.bibliographiccitation.volume","122"],["dc.contributor.author","Belov, Vladimir N."],["dc.contributor.author","Wurm, Christian Andreas"],["dc.contributor.author","Boyarskiy, Vadim P."],["dc.contributor.author","Jakobs, Stefan"],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2017-09-07T11:53:02Z"],["dc.date.available","2017-09-07T11:53:02Z"],["dc.date.issued","2010"],["dc.identifier.doi","10.1002/ange.201000150"],["dc.identifier.gro","3145021"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/2711"],["dc.language.iso","de"],["dc.notes.intern","Crossref Import"],["dc.notes.status","final"],["dc.relation.issn","0044-8249"],["dc.title","Rhodamine NN: eine neue Klasse maskierter Fluoreszenzfarbstoffe"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dc.type.peerReviewed","no"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","3593"],["dc.bibliographiccitation.issue","19"],["dc.bibliographiccitation.journal","European Journal of Organic Chemistry"],["dc.bibliographiccitation.lastpage","3610"],["dc.bibliographiccitation.volume","2010"],["dc.contributor.author","Kolmakov, Kirill"],["dc.contributor.author","Belov, Vladimir N."],["dc.contributor.author","Wurm, Christian Andreas"],["dc.contributor.author","Harke, Benjamin"],["dc.contributor.author","Leutenegger, Marcel"],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2017-09-07T11:45:57Z"],["dc.date.available","2017-09-07T11:45:57Z"],["dc.date.issued","2010"],["dc.description.abstract","Biological microscopy favors photostable fluorescent markers with large fluorescence quantum yields, low dark triplet state population, good biocompatibility and absorption and emission maxima in the near-infrared, where cellular autofluorescence is minimized. In the present study, carbopyronines absorbing around 640 nm and emitting at around 660 nm, with a low intersystem crossing rate (k(isc) approximate to 0.5 X 10(6) s(-1)) and excellent properties for cellular imaging were synthesized. A general synthetic route to carbopyronines with functional groups variable in the final steps of the synthesis or in the resulting fluorescent dye is presented. Possessing two 2-methoxyethyl groups, the parent dye is soluble in water and most organic solvents. Demethylation of the dye or its precursors is straightforward, clean, and furnishes compounds with one or two 2-hydroxyethyl groups, which can be used for further transformations. Modifications in the linker-containing carboxy group are also possible. A multistep synthesis of the dye starting from a simple precursor and utilizing a single temporary protective group is described. The presented approach may be further applied to the design of caged carbopyronines."],["dc.identifier.doi","10.1002/ejoc.201000343"],["dc.identifier.gro","3142895"],["dc.identifier.isi","000280220800007"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/350"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10 / Funder: Deutsche Forschungsgemeinschaft (DFG); Max-Planck-Gesellschaft"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","1434-193X"],["dc.title","A Versatile Route to Red-Emitting Carbopyronine Dyes for Optical Microscopy and Nanoscopy"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","690"],["dc.bibliographiccitation.issue","4"],["dc.bibliographiccitation.journal","Bioconjugate Chemistry"],["dc.bibliographiccitation.lastpage","700"],["dc.bibliographiccitation.volume","24"],["dc.contributor.author","Sednev, Maksim V."],["dc.contributor.author","Wurm, Christian Andreas"],["dc.contributor.author","Belov, Vladimir N."],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2017-09-07T11:47:44Z"],["dc.date.available","2017-09-07T11:47:44Z"],["dc.date.issued","2013"],["dc.description.abstract","Asymmetric hybrid fluorophores are built from the structural elements of two (or even more) symmetric dyes and can develop valuable new features which their parents do not possess. A new hybrid carborhodol dye was obtained by the combination of fluorescein and carbopyronine fluorophores. The brightly fluorescent hybrid dye with a linker and reactive group was prepared in 12 steps with overall yield of 1.6%. In aqueous solutions, it has absorption and emission maxima at 586 and 613 nm, respectively. Antibodies labeled with a carborhodol dye possess broad absorption and emission bands so that the effective Stokes shift is increased (compared with small Stokes shifts of the parent dyes) and the fluorescence quantum yield of 39% at a degree of labeling of 5.2. Two samples of secondary antibodies labeled with carborhodol and the benchmark red-emitting rhodamine dye (KK114) were used in two-color imaging experiments with excitation at 514-532 (carborhodol dye) and 633-640 nm (KK114). When emitted light was detected above 650 nm, the novel carborhodol dye provided a lower crosstalk than spectrally similar emitters (e. g., Atto594; crosstalk 40-60% with KK114 under the same conditions). The optical resolution of ca. 80 nm was attained using the new dye in stimulated emission depleted (STED) microscopy. The relatively short fluorescence lifetime in conjugates with antibodies (tau = 1.2-1.6 ns) suggests the possibility of dual FLIM with numerous dyes having values in the range of 3-5 ns. All of these features make the carborhodol fluorophore a valuable addition to the family of the red-emitting fluorescent dyes."],["dc.identifier.doi","10.1021/bc3006732"],["dc.identifier.gro","3142364"],["dc.identifier.isi","000317872900020"],["dc.identifier.pmid","23517127"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/7464"],["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","1043-1802"],["dc.title","Carborhodol: A New Hybrid Fluorophore Obtained by Combination of Fluorescein and Carbopyronine Dye Cores"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.artnumber","7"],["dc.bibliographiccitation.firstpage","1"],["dc.bibliographiccitation.issue","1"],["dc.bibliographiccitation.journal","Optical Nanoscopy"],["dc.bibliographiccitation.lastpage","7"],["dc.bibliographiccitation.volume","1"],["dc.contributor.author","Wurm, Christian Andreas"],["dc.contributor.author","Kolmakov, Kirill"],["dc.contributor.author","Göttfert, Fabian"],["dc.contributor.author","Ta, Haisen"],["dc.contributor.author","Bossi, Mariano"],["dc.contributor.author","Schill, Heiko"],["dc.contributor.author","Berning, Sebastian"],["dc.contributor.author","Jakobs, Stefan"],["dc.contributor.author","Donnert, Gerald"],["dc.contributor.author","Belov, Vladimir N."],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2017-09-07T11:53:03Z"],["dc.date.available","2017-09-07T11:53:03Z"],["dc.date.issued","2012"],["dc.description.abstract","In optical microscopy, most red-emitting dyes provide only moderate performance due to unspecific binding, poor labeling efficiency, and insufficient brightness. Here we report on four novel red fluororescent dyes, including the first phosphorylated dye, created by combining a rigidized rhodamine backbone with various polar groups. They exhibit large fluorescence quantum yields and improved NHS ester stability. While these fluorophores are highly suitable for fluorescence microscopy in general, they excel in stimulated emission depletion (STED) microscopy, providing < 25 nm spatial resolution in raw images of cells."],["dc.identifier.doi","10.1186/2192-2853-1-7"],["dc.identifier.fs","593636"],["dc.identifier.gro","3145019"],["dc.identifier.purl","https://resolver.sub.uni-goettingen.de/purl?gs-1/8898"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/2709"],["dc.language.iso","en"],["dc.notes.intern","Crossref Import"],["dc.notes.intern","Merged from goescholar"],["dc.notes.status","final"],["dc.relation.issn","2192-2853"],["dc.relation.orgunit","Fakultät für Physik"],["dc.rights","Goescholar"],["dc.rights.uri","https://goedoc.uni-goettingen.de/licenses"],["dc.title","Novel red fluorophores with superior performance in STED microscopy"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","no"],["dc.type.version","published_version"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","13162"],["dc.bibliographiccitation.issue","41"],["dc.bibliographiccitation.journal","Chemistry - A European Journal"],["dc.bibliographiccitation.lastpage","13173"],["dc.bibliographiccitation.volume","20"],["dc.contributor.author","Belov, Vladimir N."],["dc.contributor.author","Mitronova, Gyuzel Yu"],["dc.contributor.author","Bossi, Mariano L."],["dc.contributor.author","Boyarskiy, Vadim P."],["dc.contributor.author","Hebisch, Elke"],["dc.contributor.author","Geisler, Claudia"],["dc.contributor.author","Kolmakov, Kirill"],["dc.contributor.author","Wurm, Christian Andreas"],["dc.contributor.author","Willig, Katrin I."],["dc.contributor.author","Hell, Stefan"],["dc.date.accessioned","2017-09-07T11:45:27Z"],["dc.date.available","2017-09-07T11:45:27Z"],["dc.date.issued","2014"],["dc.description.abstract","Caged rhodamine dyes (Rhodamines NN) of five basic colors were synthesized and used as \"hidden\" markers in subdiffractional and conventional light microscopy. These masked fluorophores with a 2-diazo-1-indanone group can be irreversibly photoactivated, either by irradiation with UV- or violet light (one-photon process), or by exposure to intense red light (lambda-750 nm; two-photon mode). All dyes possess a very small 2-diazoketone caging group incorporated into the 2-diazo-1-indanone residue with a quaternary carbon atom (C-3) and a spiro-9H-xanthene fragment. Initially they are non-colored (pale yellow), non-fluorescent, and absorb at lambda=330-350 nm (molar extinction coefficient (epsilon) approximate to 10(4)M(-1)cm(-1)) with a band edge that extends to about lambda = 440 nm. The absorption and emission bands of the uncaged derivatives are tunable over a wide range (lambda=511-633 and 525-653 nm, respectively). The unmasked dyes are highly colored and fluorescent (epsilon = 3-8 x 10(4)M(-1)cm(-1) and fluorescence quantum yields (phi) = 40-85% in the unbound state and in methanol). By stepwise and orthogonal protection of carboxylic and sulfonic acid groups a highly water-soluble caged red-emitting dye with two sulfonic acid residues was prepared. Rhodamines NN were decorated with amino-reactive N-hydroxysuccinimidyl ester groups, applied in aqueous buffers, easily conjugated with proteins, and readily photoactivated (uncaged) with lambda = 375-420 nm light or intense red light (lambda = 775 nm). Protein conjugates with optimal degrees of labeling (3-6) were prepared and uncaged with lambda= 405 nm light in aqueous buffer solutions (phi = 20-38%). The photochemical cleavage of the masking group generates only molecular nitrogen. Some 10-40% of the non-fluorescent (dark) byproducts are also formed. However, they have low absorbance and do not quench the fluorescence of the uncaged dyes. Photoactivation of the individual molecules of Rhodamines NN (e.g., due to reversible or irreversible transition to a \"dark\" non-emitting state or photobleaching) provides multicolor images with subdiffractional optical resolution. The applicability of these novel caged fluorophores in super-resolution optical microscopy is exemplified."],["dc.identifier.doi","10.1002/chem.201403316"],["dc.identifier.gro","3142036"],["dc.identifier.isi","000342770900020"],["dc.identifier.pmid","25196166"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/3834"],["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","1521-3765"],["dc.relation.issn","0947-6539"],["dc.title","Masked Rhodamine Dyes of Five Principal Colors Revealed by Photolysis of a 2-Diazo-1-Indanone Caging Group: Synthesis, Photophysics, and Light Microscopy Applications"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","146"],["dc.bibliographiccitation.issue","1"],["dc.bibliographiccitation.journal","Chemistry - A European Journal"],["dc.bibliographiccitation.lastpage","157"],["dc.bibliographiccitation.volume","20"],["dc.contributor.author","Kolmakov, Kirill"],["dc.contributor.author","Wurm, Christian Andreas"],["dc.contributor.author","Meineke, Dirk N. H."],["dc.contributor.author","Göttfert, Fabian"],["dc.contributor.author","Boyarskiy, Vadim P."],["dc.contributor.author","Belov, Vladimir N."],["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","The synthesis, reactivity, and photophysical properties of new rhodamines with intense red fluorescence, two polar residues (hydroxyls, primary phosphates, or sulfonic acid groups), and improved hydrolytic stability of the amino-reactive sites (NHS esters or mixed N-succinimidyl carbonates) are reported. All fluorophores contain an N-alkyl-1,2-dihydro-2,2,4-trimethylquinoline fragment, and most of them bear a fully substituted tetrafluoro phenyl ring with a secondary carboxamide group. The absorption and emission maxima in water are in the range of 635-639 and 655-659nm, respectively. A vastly simplified approach to red-emitting rhodamines with two phosphate groups that are compatible with diverse functional linkers was developed. As an example, a phosphorylated dye with an azide residue was prepared and was used in a click reaction with a strained alkyne bearing an N-hydroxysuccinimid (NHS) ester group. This method bypasses the undesired activation of phosphate groups, and gives an amphiphilic amino-reactive dye, the solubility and distribution of which between aqueous and organic phases can be controlled by varying the pH. The presence of two hydroxyl groups and a phenyl ring with two carboxyl residues in the dyes with another substitution pattern is sufficient for providing the hydrophilic properties. Selective formation of a mono-N-hydroxysuccinimidyl ester from 5-carboxy isomer of this rhodamine is reported. The fluorescence quantum yields varied from 58 to 92% for free fluorophores, and amounted to 18-64% for antibody conjugates in aqueous buffers. The brightness and photostability of these fluorophores facilitated two-color stimulated emission depletion (STED) fluorescence nanoscopy of biological samples with high contrast and minimal background. Selecting a pair of fluorophores with absorption/emission bands at 579/609 and 635/655nm enabled two-color channels with low cross-talk and negligible background at approximately 40nm resolution."],["dc.identifier.doi","10.1002/chem.201303433"],["dc.identifier.gro","3142200"],["dc.identifier.isi","000328714700019"],["dc.identifier.pmid","24338798"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/5643"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10 / Funder: Bundesministerium fur Bildung und Forschung [BMBF 513, FKZ 13N11066]"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.eissn","1521-3765"],["dc.relation.issn","0947-6539"],["dc.title","Polar Red-Emitting Rhodamine Dyes with Reactive Groups: Synthesis, Photophysical Properties, and Two-Color STED Nanoscopy Applications"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","12986"],["dc.bibliographiccitation.issue","41"],["dc.bibliographiccitation.journal","Chemistry - A European Journal"],["dc.bibliographiccitation.lastpage","12998"],["dc.bibliographiccitation.volume","18"],["dc.contributor.author","Kolmakov, Kirill"],["dc.contributor.author","Wurm, Christian Andreas"],["dc.contributor.author","Hennig, René"],["dc.contributor.author","Rapp, Erdmann"],["dc.contributor.author","Jakobs, Stefan"],["dc.contributor.author","Belov, Vladimir N."],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2017-09-07T11:48:24Z"],["dc.date.available","2017-09-07T11:48:24Z"],["dc.date.issued","2012"],["dc.description.abstract","Fluorescent dyes emitting red light are frequently used in conventional and super-resolution microscopy of biological samples, although the variety of the useful dyes is limited. We describe the synthesis of rhodamine-based fluorescent dyes with absorption and emission maxima in the range of 621637 and 644660 nm, respectively and demonstrate their high performance in confocal and stimulated emission depletion (STED) microscopy. New dyes were prepared by means of reliable chemical transformations applied to a rhodamine scaffold with three variable positions. They feature polarity, water solubility, variable net charges, improved stabilities of N-hydroxysuccinimidyl (NHS) esters, as well as large fluorescence quantum yields in dye solutions and antibody conjugates. The photophysical and imaging properties of dyes containing three different polar groups, namely primary phosphate, sulfonic acid (in two different positions), and hydroxyl were compared. A dye with two primary phosphate groups was explored as a valuable alternative to dyes with classical sulfonic acid groups. Due to the increased net charge of the phosphorylated dye (q=-4 at pH 8), it demonstrated a far better electrophoretic mobility compared with analogues with two sulfonic acid groups (q=-2). As an example, one fluorescent dye was designed to be especially convenient for practical use. It is characterized by sufficiently high chemical stability of the NHS ester, its simple isolation, handling, and solubility in aqueous buffers, as well as in organic solvents. All these features, accompanied by a zero net charge in conjugates, were accomplished by the introduction of hydrophilic groups of two types: two hydroxyl groups and one sulfonic acid residue."],["dc.identifier.doi","10.1002/chem.201201168"],["dc.identifier.gro","3142458"],["dc.identifier.isi","000309239300013"],["dc.identifier.pmid","22968960"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/8507"],["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","0947-6539"],["dc.title","Red-Emitting Rhodamines with Hydroxylated, Sulfonated, and Phosphorylated Dye Residues and Their Use in Fluorescence Nanoscopy"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.artnumber","e78745"],["dc.bibliographiccitation.issue","10"],["dc.bibliographiccitation.journal","PLOS ONE"],["dc.bibliographiccitation.volume","8"],["dc.contributor.author","Stagge, Franziska"],["dc.contributor.author","Mitronova, Gyuzel Yu"],["dc.contributor.author","Belov, Vladimir N."],["dc.contributor.author","Wurm, Christian Andreas"],["dc.contributor.author","Jakobs, Stefan"],["dc.date.accessioned","2017-09-07T11:47:06Z"],["dc.date.available","2017-09-07T11:47:06Z"],["dc.date.issued","2013"],["dc.description.abstract","Fluorescence microscopy of the localization and the spatial and temporal dynamics of specifically labelled proteins is an indispensable tool in cell biology. Besides fluorescent proteins as tags, tag-mediated labelling utilizing self-labelling proteins as the SNAP-, CLIP-, or the Halo-tag are widely used, flexible labelling systems relying on exogenously supplied fluorophores. Unfortunately, labelling of live budding yeast cells proved to be challenging with these approaches because of the limited accessibility of the cell interior to the dyes. In this study we developed a fast and reliable electroporation-based labelling protocol for living budding yeast cells expressing SNAP-, CLIP-, or Halo-tagged fusion proteins. For the Halo-tag, we demonstrate that it is crucial to use the 6'-carboxy isomers and not the 5'-carboxy isomers of important dyes to ensure cell viability. We report on a simple rule for the analysis of H-1 NMR spectra to discriminate between 6'- and 5'-carboxy isomers of fluorescein and rhodamine derivatives. We demonstrate the usability of the labelling protocol by imaging yeast cells with STED super-resolution microscopy and dual colour live cell microscopy. The large number of available fluorophores for these self-labelling proteins and the simplicity of the protocol described here expands the available toolbox for the model organism Saccharomyces cerevisiae."],["dc.identifier.doi","10.1371/journal.pone.0078745"],["dc.identifier.gro","3142268"],["dc.identifier.isi","000326155400102"],["dc.identifier.pmid","24205303"],["dc.identifier.purl","https://resolver.sub.uni-goettingen.de/purl?gs-1/9435"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/6398"],["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.issn","1932-6203"],["dc.rights","CC BY 2.5"],["dc.rights.uri","https://creativecommons.org/licenses/by/2.5"],["dc.title","Snap-, CLIP- and Halo-Tag Labelling of Budding Yeast Cells"],["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"]]