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A Co-Culture-Based Multiparametric Imaging Technique to Dissect Local H2O2 Signals with Targeted HyPer7

Secilmis, Melike; Altun, Hamza Yusuf; Pilic, Johannes; Erdogan, Yusuf Ceyhun; Cokluk, Zeynep; Ata, Busra Nur; Sevimli, Gulsah; Zaki, Asal Ghaffari; Yigit, Esra Nur; Ozturk, Gurkan; Malli, Roland; Eroglu, Emrah


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  <identifier identifierType="URL">https://aperta.ulakbim.gov.tr/record/237518</identifier>
  <creators>
    <creator>
      <creatorName>Secilmis, Melike</creatorName>
      <givenName>Melike</givenName>
      <familyName>Secilmis</familyName>
      <affiliation>Sabanci Univ, Mol Biol Genet &amp; Bioengn Program, TR-34956 Istanbul, Turkey</affiliation>
    </creator>
    <creator>
      <creatorName>Altun, Hamza Yusuf</creatorName>
      <givenName>Hamza Yusuf</givenName>
      <familyName>Altun</familyName>
      <affiliation>Sabanci Univ, Mol Biol Genet &amp; Bioengn Program, TR-34956 Istanbul, Turkey</affiliation>
    </creator>
    <creator>
      <creatorName>Pilic, Johannes</creatorName>
      <givenName>Johannes</givenName>
      <familyName>Pilic</familyName>
      <affiliation>Med Univ Graz, Gottfried Schatz Res Ctr, Mol Biol &amp; Biochem, A-8036 Graz, Austria</affiliation>
    </creator>
    <creator>
      <creatorName>Erdogan, Yusuf Ceyhun</creatorName>
      <givenName>Yusuf Ceyhun</givenName>
      <familyName>Erdogan</familyName>
    </creator>
    <creator>
      <creatorName>Cokluk, Zeynep</creatorName>
      <givenName>Zeynep</givenName>
      <familyName>Cokluk</familyName>
      <affiliation>Sabanci Univ, Mol Biol Genet &amp; Bioengn Program, TR-34956 Istanbul, Turkey</affiliation>
    </creator>
    <creator>
      <creatorName>Ata, Busra Nur</creatorName>
      <givenName>Busra Nur</givenName>
      <familyName>Ata</familyName>
      <affiliation>Sabanci Univ, Mol Biol Genet &amp; Bioengn Program, TR-34956 Istanbul, Turkey</affiliation>
    </creator>
    <creator>
      <creatorName>Sevimli, Gulsah</creatorName>
      <givenName>Gulsah</givenName>
      <familyName>Sevimli</familyName>
      <affiliation>Sabanci Univ, Mol Biol Genet &amp; Bioengn Program, TR-34956 Istanbul, Turkey</affiliation>
    </creator>
    <creator>
      <creatorName>Zaki, Asal Ghaffari</creatorName>
      <givenName>Asal Ghaffari</givenName>
      <familyName>Zaki</familyName>
      <affiliation>Sabanci Univ, Mol Biol Genet &amp; Bioengn Program, TR-34956 Istanbul, Turkey</affiliation>
    </creator>
    <creator>
      <creatorName>Yigit, Esra Nur</creatorName>
      <givenName>Esra Nur</givenName>
      <familyName>Yigit</familyName>
    </creator>
    <creator>
      <creatorName>Ozturk, Gurkan</creatorName>
      <givenName>Gurkan</givenName>
      <familyName>Ozturk</familyName>
    </creator>
    <creator>
      <creatorName>Malli, Roland</creatorName>
      <givenName>Roland</givenName>
      <familyName>Malli</familyName>
    </creator>
    <creator>
      <creatorName>Eroglu, Emrah</creatorName>
      <givenName>Emrah</givenName>
      <familyName>Eroglu</familyName>
    </creator>
  </creators>
  <titles>
    <title>A Co-Culture-Based Multiparametric Imaging Technique To Dissect Local H2O2 Signals With Targeted Hyper7</title>
  </titles>
  <publisher>Aperta</publisher>
  <publicationYear>2021</publicationYear>
  <dates>
    <date dateType="Issued">2021-01-01</date>
  </dates>
  <resourceType resourceTypeGeneral="Text">Journal article</resourceType>
  <alternateIdentifiers>
    <alternateIdentifier alternateIdentifierType="url">https://aperta.ulakbim.gov.tr/record/237518</alternateIdentifier>
  </alternateIdentifiers>
  <relatedIdentifiers>
    <relatedIdentifier relatedIdentifierType="DOI" relationType="IsIdenticalTo">10.3390/bios11090338</relatedIdentifier>
  </relatedIdentifiers>
  <rightsList>
    <rights rightsURI="http://www.opendefinition.org/licenses/cc-by">Creative Commons Attribution</rights>
    <rights rightsURI="info:eu-repo/semantics/openAccess">Open Access</rights>
  </rightsList>
  <descriptions>
    <description descriptionType="Abstract">Multispectral 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.</description>
  </descriptions>
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