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SHR Neuro Krebs Kardio Lipid Stoffw Microb

Secilmis, M; Altun, HY; Pilic, J; Erdogan, YC; Cokluk, Z; Ata, BN; Sevimli, G; Zaki, AG; Yigit, EN; Öztürk, G; Malli, R; Eroglu, E.
A Co-Culture-Based Multiparametric Imaging Technique to Dissect Local H2O2 Signals with Targeted HyPer7.
Biosensors (Basel). 2021; 11(9): 338 Doi: 10.3390/bios11090338 [OPEN ACCESS]
Web of Science PubMed PUBMED Central FullText FullText_MUG

 

Führende Autor*innen der Med Uni Graz
EROGLU Emrah
Malli Roland
Co-Autor*innen der Med Uni Graz
Erdogan Yusuf Ceyhun
Pilic Johannes
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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.

Find related publications in this database (Keywords)
endothelial cell line
HyPer7
hydrogen peroxide
targeted biosensors
co-imaging
mitochondria
multiparametric imaging
multiplex live-cell imaging
stable cell lines
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