Testa, Ilaria

[["dc.bibliographiccitation.artnumber","9592"],["dc.bibliographiccitation.journal","Scientific Reports"],["dc.bibliographiccitation.volume","5"],["dc.contributor.author","Ratz, Michael"],["dc.contributor.author","Testa, Ilaria"],["dc.contributor.author","Hell, Stefan W."],["dc.contributor.author","Jakobs, Stefan"],["dc.date.accessioned","2017-09-07T11:44:27Z"],["dc.date.available","2017-09-07T11:44:27Z"],["dc.date.issued","2015"],["dc.description.abstract","Overexpression is a notorious concern in conventional and especially in super-resolution fluorescence light microscopy studies because it may cause numerous artifacts including ectopic sub-cellular localizations, erroneous formation of protein complexes, and others. Nonetheless, current live cell super-resolution microscopy studies generally rely on the overexpression of a host protein fused to a fluorescent protein. Here, we establish CRISPR/Cas9-mediated generation of heterozygous and homozygous human knockin cell lines expressing fluorescently tagged proteins from their respective native genomic loci at close to endogenous levels. We tagged three different proteins, exhibiting various localizations and expression levels, with the reversibly switchable fluorescent protein rsEGFP2. We demonstrate the benefit of endogenous expression levels compared to overexpression and show that typical overexpression-induced artefacts were avoided in genome-edited cells. Fluorescence activated cell sorting analysis revealed a narrow distribution of fusion protein expression levels in genome-edited cells, compared to a pronounced variability in transiently transfected cells. Using low light intensity RESOLFT (reversible saturable optical fluorescence transitions) nanoscopy we show sub-diffraction resolution imaging of living human knockin cells. Our strategy to generate human cell lines expressing fluorescent fusion proteins at endogenous levels for RESOLFT nanoscopy can be extended to other fluorescent tags and super-resolution approaches."],["dc.identifier.doi","10.1038/srep09592"],["dc.identifier.gro","3141924"],["dc.identifier.isi","000353276500001"],["dc.identifier.pmid","25892259"],["dc.identifier.purl","https://resolver.sub.uni-goettingen.de/purl?gs-1/13621"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/2589"],["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","2045-2322"],["dc.rights","CC BY 4.0"],["dc.rights.uri","https://creativecommons.org/licenses/by/4.0"],["dc.title","CRISPR/Cas9-mediated endogenous protein tagging for RESOLFT super-resolution microscopy of living human 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"]]

[["dc.bibliographiccitation.firstpage","655"],["dc.bibliographiccitation.issue","4"],["dc.bibliographiccitation.journal","ChemPhysChem"],["dc.bibliographiccitation.lastpage","663"],["dc.bibliographiccitation.volume","15"],["dc.contributor.author","Lavoie-Cardinal, Flavie"],["dc.contributor.author","Jensen, Nickels A."],["dc.contributor.author","Westphal, Volker"],["dc.contributor.author","Stiel, Andre C."],["dc.contributor.author","Chmyrov, Andriy"],["dc.contributor.author","Bierwagen, Jakob"],["dc.contributor.author","Testa, Ilaria"],["dc.contributor.author","Jakobs, Stefan"],["dc.contributor.author","Hell, Stefan W."],["dc.date.accessioned","2017-09-07T11:46:25Z"],["dc.date.available","2017-09-07T11:46:25Z"],["dc.date.issued","2014"],["dc.description.abstract","Up to now, all demonstrations of reversible saturable optical fluorescence transitions (RESOLFT) superresolution microscopy of living cells have relied on the use of reversibly switchable fluorescent proteins (RSFP) emitting in the green spectral range. Here we show RESOLFT imaging with rsCherryRev1.4, a new red-emitting RSFP enabling a spatial resolution up to four times higher than the diffraction barrier. By co-expressing green and red RSFPs in living cells we demonstrate two-color RESOLFT imaging both for single (donut) beam scanning and for parallelized versions of RESOLFT nanoscopy where an array of >23000 donut-like minima are scanned simultaneously."],["dc.identifier.doi","10.1002/cphc.201301016"],["dc.identifier.gro","3142168"],["dc.identifier.isi","000332747500013"],["dc.identifier.pmid","24449030"],["dc.identifier.purl","https://resolver.sub.uni-goettingen.de/purl?gs-1/12826"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/5288"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10 / Funder: Deutsche Forschungsgemeinschaft"],["dc.notes.intern","Merged from goescholar"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.eissn","1439-7641"],["dc.relation.issn","1439-4235"],["dc.rights","CC BY-NC-ND 3.0"],["dc.rights.uri","https://creativecommons.org/licenses/by-nc-nd/3.0"],["dc.title","Two-Color RESOLFT Nanoscopy with Green and Red Fluorescent Photochromic Proteins"],["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","e00248"],["dc.bibliographiccitation.firstpage","1"],["dc.bibliographiccitation.journal","eLife"],["dc.bibliographiccitation.lastpage","14"],["dc.bibliographiccitation.volume","1"],["dc.contributor.author","Grotjohann, Tim"],["dc.contributor.author","Testa, Ilaria"],["dc.contributor.author","Reuss, Matthias"],["dc.contributor.author","Brakemann, Tanja"],["dc.contributor.author","Eggeling, Christian"],["dc.contributor.author","Hell, Stefan W."],["dc.contributor.author","Jakobs, Stefan"],["dc.date.accessioned","2017-09-07T11:48:20Z"],["dc.date.available","2017-09-07T11:48:20Z"],["dc.date.issued","2012"],["dc.description.abstract","The super-resolution microscopy called RESOLFT relying on fluorophore switching between longlived states, stands out by its coordinate-targeted sequential sample interrogation using low light levels. While RESOLFT has been shown to discern nanostructures in living cells, the reversibly photoswitchable green fluorescent protein (rsEGFP) employed in these experiments was switched rather slowly and recording lasted tens of minutes. We now report on the generation of rsEGFP2 providing faster switching and the use of this protein to demonstrate 25-250 times faster recordings."],["dc.identifier.doi","10.7554/eLife.00248"],["dc.identifier.gro","3142424"],["dc.identifier.isi","000328585000001"],["dc.identifier.pmid","23330067"],["dc.identifier.purl","https://resolver.sub.uni-goettingen.de/purl?gs-1/10590"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/8130"],["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","2050-084X"],["dc.rights","CC BY 3.0"],["dc.rights.uri","https://creativecommons.org/licenses/by/3.0"],["dc.title","rsEGFP2 enables fast RESOLFT nanoscopy of living 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"]]

[["dc.bibliographiccitation.firstpage","139"],["dc.bibliographiccitation.issue","1"],["dc.bibliographiccitation.journal","The EMBO journal"],["dc.bibliographiccitation.lastpage","159"],["dc.bibliographiccitation.volume","37"],["dc.contributor.author","Richter, Katharina N."],["dc.contributor.author","Revelo, Natalia H."],["dc.contributor.author","Seitz, Katharina J."],["dc.contributor.author","Helm, Martin S."],["dc.contributor.author","Sarkar, Deblina"],["dc.contributor.author","Saleeb, Rebecca S."],["dc.contributor.author","d'Este, Elisa"],["dc.contributor.author","Eberle, Jessica"],["dc.contributor.author","Wagner, Eva"],["dc.contributor.author","Vogl, Christian"],["dc.contributor.author","Lazaro, Diana F."],["dc.contributor.author","Richter, Frank"],["dc.contributor.author","Coy-Vergara, Javier"],["dc.contributor.author","Coceano, Giovanna"],["dc.contributor.author","Boyden, Edward S."],["dc.contributor.author","Duncan, Rory R."],["dc.contributor.author","Hell, Stefan W."],["dc.contributor.author","Lauterbach, Marcel A."],["dc.contributor.author","Lehnart, Stephan E."],["dc.contributor.author","Moser, Tobias"],["dc.contributor.author","Outeiro, Tiago F."],["dc.contributor.author","Rehling, Peter"],["dc.contributor.author","Schwappach, Blanche"],["dc.contributor.author","Testa, Ilaria"],["dc.contributor.author","Zapiec, Bolek"],["dc.contributor.author","Rizzoli, Silvio O."],["dc.date.accessioned","2018-01-09T14:09:53Z"],["dc.date.available","2018-01-09T14:09:53Z"],["dc.date.issued","2018"],["dc.description.abstract","Paraformaldehyde (PFA) is the most commonly used fixative for immunostaining of cells, but has been associated with various problems, ranging from loss of antigenicity to changes in morphology during fixation. We show here that the small dialdehyde glyoxal can successfully replace PFA Despite being less toxic than PFA, and, as most aldehydes, likely usable as a fixative, glyoxal has not yet been systematically tried in modern fluorescence microscopy. Here, we tested and optimized glyoxal fixation and surprisingly found it to be more efficient than PFA-based protocols. Glyoxal acted faster than PFA, cross-linked proteins more effectively, and improved the preservation of cellular morphology. We validated glyoxal fixation in multiple laboratories against different PFA-based protocols and confirmed that it enabled better immunostainings for a majority of the targets. Our data therefore support that glyoxal can be a valuable alternative to PFA for immunostaining."],["dc.identifier.doi","10.15252/embj.201695709"],["dc.identifier.pmid","29146773"],["dc.identifier.purl","https://resolver.sub.uni-goettingen.de/purl?gs-1/15063"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/11599"],["dc.identifier.url","https://sfb1002.med.uni-goettingen.de/production/literature/publications/195"],["dc.identifier.url","https://sfb1190.med.uni-goettingen.de/production/literature/publications/15"],["dc.language.iso","en"],["dc.notes.intern","Merged from goescholar"],["dc.notes.status","final"],["dc.relation","SFB 1002: Modulatorische Einheiten bei Herzinsuffizienz"],["dc.relation","SFB 1002 | A09: Lokale molekulare Nanodomänen-Regulation der kardialen Ryanodin-Rezeptor-Funktion"],["dc.relation","SFB 1190: Transportmaschinen und Kontaktstellen zellulärer Kompartimente"],["dc.relation","SFB 1190 | P09: Proteinsortierung in der Synapse: Prinzipien und molekulare Organisation"],["dc.relation.eissn","1460-2075"],["dc.relation.workinggroup","RG Hell"],["dc.relation.workinggroup","RG Lehnart (Cellular Biophysics and Translational Cardiology Section)"],["dc.relation.workinggroup","RG Rehling (Mitochondrial Protein Biogenesis)"],["dc.relation.workinggroup","RG Schwappach (Membrane Protein Biogenesis)"],["dc.relation.workinggroup","RG Rizzoli (Quantitative Synaptology in Space and Time)"],["dc.rights","CC BY 4.0"],["dc.rights.uri","https://creativecommons.org/licenses/by/4.0"],["dc.title","Glyoxal as an alternative fixative to formaldehyde in immunostaining and super-resolution microscopy"],["dc.type","journal_article"],["dc.type.internalPublication","unknown"],["dc.type.subtype","original_ja"],["dc.type.version","published_version"],["dspace.entity.type","Publication"]]