New generation of fluorescent probes for cell-based measurements of caspase Activation and mitochondrial superoxide

Abstract Cell health and stress readouts are critical indicators of altered or impaired function in normal diseased states cells, work has been underway to develop improved small molecule sensor dyes compatible with traditional imaging High Content Analysis (HCA) interrogation apoptotic mitochondrial pathways. The CellEvent™ Caspase Green dye effectively reports caspase activation, but suffers complications assay configuration when attempting multiplex the Fluorescent Protein (GFP), calcein, other 488 laser line tools fluorescence microscopy. Here, we describe testing functional characterization a new candidate for measuring apoptosis living cells. Our is comprised fluorogenic reporter that liberated from DEVD peptide substrate by ex/em spectra 590/610 nm, permitting easy GFP calcein stained neurons both HCA microscopy configurations. MitoSOX™ specificity superoxide like Red dye. In addition, it better spectral properties works filters microscopy, allows measurement RFP-expressing Mitochondrial Superoxide Indicator also localizes mitochondria live cells selectively generation, while ignoring ROS RNS species ex vivo testing. With an Excitation/Emission profile GFP/FITC channel, series comparative studies immortalized neural shown, highlighting photostability, signal amplitude These reagents research use only, not diagnostic purposes