A co-culture-based multiparametric imaging technique to dissect local H2O2 signals with targeted HyPer7

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dc.authorid0000-0001-9037-5217
dc.authorid0000-0003-0352-1947
dc.authorid0000-0002-9373-0808
dc.contributor.authorSeçilmiş, Melike
dc.contributor.authorAltun, Hamza Yusuf
dc.contributor.authorPilic, Johannes
dc.contributor.authorErdoğan, Yusuf Ceyhun
dc.contributor.authorÇokluk, Zeynep
dc.contributor.authorAta, Büşra Nur
dc.contributor.authorSevimli, Gülşah
dc.contributor.authorZaki, Asal Ghaffari
dc.contributor.authorYiğit, Esra Nur
dc.contributor.authorÖztürk, Gürkan
dc.contributor.authorMalli, Roland
dc.contributor.authorEroğlu, Emrah
dc.date.accessioned2021-10-07T07:17:27Z
dc.date.available2021-10-07T07:17:27Z
dc.date.issued2021
dc.departmentİstanbul Medipol Üniversitesi, Rektörlük, Sağlık Bilim ve Teknolojileri Araştırma Enstitüsü
dc.departmentİstanbul Medipol Üniversitesi, Uluslararası Tıp Fakültesi, Temel Tıp Bilimleri Bölümü, Fizyoloji Ana Bilim Dalı
dc.description.abstractMultispectral live-cell imaging is an informative approach that permits detecting biological processes simultaneously in the spatial and temporal domain by exploiting spectrally distinct biosensors. However, the combination of fluorescent biosensors with distinct spectral properties such as different sensitivities, and dynamic ranges can undermine accurate co-imaging of the same analyte in different subcellular locales. We advanced a single-color multiparametric imaging method, which allows simultaneous detection of hydrogen peroxide (H2O2) in multiple cell locales (nucleus, cytosol, mitochondria) using the H2O2 biosensor HyPer7. Co-culturing of endothelial cells stably expressing differentially targeted HyPer7 biosensors paved the way for co-imaging compartmentalized H2O2 signals simultaneously in neighboring cells in a single experimental setup. We termed this approach COMPARE IT, which is an acronym for co-culture-based multiparametric imaging technique. Employing this approach, we detected lower H2O2 levels in mitochondria of endothelial cells compared to the cell nucleus and cytosol under basal conditions. Upon administering exogenous H2O2, the cytosolic and nuclear-targeted probes displayed similarly slow and moderate HyPer7 responses, whereas the mitochondria-targeted HyPer7 signal plateaued faster and reached higher amplitudes. Our results indicate striking differences in mitochondrial H2O2 accumulation of endothelial cells. Here, we present the method's potential as a practicable and informative multiparametric live-cell imaging technique.
dc.description.sponsorshipBAGEP award of the Science Academy Turkey ; Integration Projects of Sabanci University ; Austrian Science Fund (FWF) ; Medical University of Grazen_US
dc.identifier.citationSeçilmiş, M., Altun, H. Y., Pilic, J., Erdoğan, Y. C., Çokluk, Z., Ata, B. N. ... Eroğlu, E. (2021). A co-culture-based multiparametric imaging technique to dissect local H2O2 signals with targeted HyPer7. Biosensors-Basel, 11(9). https://dx.doi.org/10.3390/bios11090338
dc.identifier.doi10.3390/bios11090338
dc.identifier.issn2079-6374
dc.identifier.issue9
dc.identifier.scopusqualityQ2
dc.identifier.urihttps://dx.doi.org/10.3390/bios11090338
dc.identifier.urihttps://hdl.handle.net/20.500.12511/8380
dc.identifier.volume11
dc.identifier.wosqualityQ1
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.indekslendigikaynakPubMed
dc.language.isoen
dc.publisherMDPI
dc.relation.ispartofBiosensors-Baselen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı
dc.relation.tubitakinfo:eu-repo/grantAgreement/TUBITAK/SOBAG/118C242
dc.rightsAttribution 4.0 International*
dc.rightsinfo:eu-repo/semantics/openAccess
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/*
dc.subjectEndothelial Cell Line
dc.subjectHyPer7
dc.subjectHydrogen Peroxide
dc.subjectTargeted Biosensors
dc.subjectCo-Imaging
dc.subjectMitochondria
dc.subjectMultiparametric Imaging
dc.subjectMultiplex Live-Cell Imaging
dc.subjectStable Cell Lines
dc.titleA co-culture-based multiparametric imaging technique to dissect local H2O2 signals with targeted HyPer7
dc.typeArticle

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