scholarly journals RNA-Based Fluorescent Biosensors for Live Cell Imaging of Second Messenger Cyclic di-AMP

2015 ◽  
Vol 137 (20) ◽  
pp. 6432-6435 ◽  
Author(s):  
Colleen A. Kellenberger ◽  
Chen Chen ◽  
Aaron T. Whiteley ◽  
Daniel A. Portnoy ◽  
Ming C. Hammond
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Second Messenger ◽  
Live Cell ◽  
Fluorescent Biosensors

Live Cell Imaging Using Riboswitch–Spinach tRNA Fusions as Metabolite-Sensing Fluorescent Biosensors

2021 ◽  
pp. 121-140
Author(s):  
Sudeshna Manna ◽  
Colleen A. Kellenberger ◽  
Zachary F. Hallberg ◽  
Ming C. Hammond
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Fluorescent Biosensors

scholarly journals RNA-Based Fluorescent Biosensors for Live Cell Imaging of Second Messengers Cyclic di-GMP and Cyclic AMP-GMP

2013 ◽  
Vol 135 (13) ◽  
pp. 4906-4909 ◽  
Author(s):  
Colleen A. Kellenberger ◽  
Stephen C. Wilson ◽  
Jade Sales-Lee ◽  
Ming C. Hammond
Keyword(s):  
Cyclic Amp ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Second Messengers ◽  
Live Cell ◽  
Fluorescent Biosensors

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

Biosensors ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 338
Author(s):  
Melike Secilmis ◽  
Hamza Yusuf Altun ◽  
Johannes Pilic ◽  
Yusuf Ceyhun Erdogan ◽  
Zeynep Cokluk ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Imaging Technique ◽  
Simultaneous Detection ◽  
Live Cell ◽  
Imaging Method ◽  
Fluorescent Biosensors ◽  
Multiparametric Imaging ◽  
Multiple Cell

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.

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