Hilbert, Michael

[["dc.bibliographiccitation.firstpage","1003"],["dc.bibliographiccitation.issue","12"],["dc.bibliographiccitation.journal","Microscopy research and technique"],["dc.bibliographiccitation.lastpage","1009"],["dc.bibliographiccitation.volume","70"],["dc.contributor.author","Eggeling, C."],["dc.contributor.author","Hilbert, M."],["dc.contributor.author","Bock, H."],["dc.contributor.author","Ringemann, C."],["dc.contributor.author","Hofmann, M."],["dc.contributor.author","Stiel, A. C."],["dc.contributor.author","Andresen, M."],["dc.contributor.author","Jakobs, S."],["dc.contributor.author","Egner, A."],["dc.contributor.author","Schönle, A."],["dc.contributor.author","Hell, S. W."],["dc.date.accessioned","2017-09-07T11:49:23Z"],["dc.date.available","2017-09-07T11:49:23Z"],["dc.date.issued","2007"],["dc.description.abstract","We demonstrate that photoswitchable markers enable fluorescence fluctuation spectroscopy at high molecular concentration. Reversible photoswitching allows precise control of the density of fluorescing entities, because the equilibrium between the fluorescent ON- and the dark OFF-state can be shifted through optical irradiation at a specific wavelength. Depending on the irradiation intensity, the concentration of the ON-state markers can be up to 1,000 times lower than the actual concentration of the labeled molecular entity. Photoswitching expands the range of single-molecule detection based experiments such as fluorescence fluctuation spectroscopy to large entity concentrations in the micromolar range."],["dc.identifier.doi","10.1002/jemt.20505"],["dc.identifier.gro","3143405"],["dc.identifier.isi","000251868200001"],["dc.identifier.pmid","17661359"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/916"],["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","1059-910X"],["dc.title","Reversible photoswitching enables single-molecule fluorescence fluctuation Spectroscopy at high molecular concentration"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","2989"],["dc.bibliographiccitation.issue","6"],["dc.bibliographiccitation.journal","Biophysical Journal"],["dc.bibliographiccitation.lastpage","2997"],["dc.bibliographiccitation.volume","95"],["dc.contributor.author","Stiel, Andre C."],["dc.contributor.author","Andresen, Martin"],["dc.contributor.author","Bock, Hannes"],["dc.contributor.author","Hilbert, Michael"],["dc.contributor.author","Schilde, Jessica"],["dc.contributor.author","Schönle, Andreas"],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Egner, Alexander"],["dc.contributor.author","Hell, Stefan"],["dc.contributor.author","Jakobs, Stefan"],["dc.date.accessioned","2017-09-07T11:48:12Z"],["dc.date.available","2017-09-07T11:48:12Z"],["dc.date.issued","2008"],["dc.description.abstract","Reversibly switchable fluorescent proteins (RSFPs) are GFP-like proteins that may be repeatedly switched by irradiation with light from a fluorescent to a nonfluorescent state, and vice versa. They can be utilized as genetically encodable probes and bear large potential for a wide array of applications, in particular for new protein tracking schemes and subdiffraction resolution microscopy. However, the currently described monomeric RSFPs emit only blue-green or green fluorescence; the spectral window for their use is thus rather limited. Using a semirational engineering approach based on the crystal structure of the monomeric nonswitchable red fluorescent protein mCherry, we generated rsCherry and rsCherryRev. These two novel red fluorescent RSFPs exhibit fluorescence emission maxima at similar to 610 nm. They display antagonistic switching modes, i.e., in rsCherry irradiation with yellow light induces the off-to-on transition and blue light the on-to-off transition, whereas in rsCherryRev the effects of the switching wavelengths are reversed. We demonstrate time-lapse live-cell subdiffraction microscopy by imaging rsCherryRev targeted to the endoplasmic reticulum utilizing the switching and localization of single molecules."],["dc.identifier.doi","10.1529/biophysj.108.130146"],["dc.identifier.gro","3143236"],["dc.identifier.isi","000258826900034"],["dc.identifier.pmid","18658221"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/728"],["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","0006-3495"],["dc.title","Generation of monomeric reversibly switchable red fluorescent proteins for far-field fluorescence nanoscopy"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","6829"],["dc.bibliographiccitation.issue","15"],["dc.bibliographiccitation.journal","Proceedings of the National Academy of Sciences"],["dc.bibliographiccitation.lastpage","6834"],["dc.bibliographiccitation.volume","107"],["dc.contributor.author","Sahl, Steffen J."],["dc.contributor.author","Leutenegger, Marcel"],["dc.contributor.author","Hilbert, Michael"],["dc.contributor.author","Hell, Stefan"],["dc.contributor.author","Eggeling, Christian"],["dc.date.accessioned","2017-09-07T11:46:06Z"],["dc.date.available","2017-09-07T11:46:06Z"],["dc.date.issued","2010"],["dc.description.abstract","We describe an optical method capable of tracking a single fluorescent molecule with a flexible choice of high spatial accuracy (similar to 10-20 nm standard deviation or similar to 20-40 nm full-width-at-half-maximum) and temporal resolution (<1 ms). The fluorescence signal during individual passages of fluorescent molecules through a spot of excitation light allows the sequential localization and thus spatio-temporal tracking of the molecule if its fluorescence is collected on at least three separate point detectors arranged in close proximity. We show two-dimensional trajectories of individual, small organic dye labeled lipids diffusing in the plasma membrane of living cells and directly observe transient events of trapping on <20 nm spatial scales. The trapping is cholesterol-assisted and much more pronounced for a sphingo- than for a phosphoglycero-lipid, with average trapping times of similar to 15 ms and <4 ms, respectively. The results support previous STED nanoscopy measurements and suggest that, at least for nontreated cells, the transient interaction of a single lipid is confined to macromolecular dimensions. Our experimental approach demonstrates that fast molecular movements can be tracked with minimal invasion, which can reveal new important details of cellular nano-organization."],["dc.identifier.doi","10.1073/pnas.0912894107"],["dc.identifier.gro","3142937"],["dc.identifier.isi","000276642100050"],["dc.identifier.pmid","20351247"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/396"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10 / Funder: Leibniz-Prize"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","0027-8424"],["dc.title","Fast molecular tracking maps nanoscale dynamics of plasma membrane lipids"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]