Ratiometric two-photon fluorescent probe for in situ imaging of carboxylesterase (CE)-mediated mitochondrial...

Hydrogen peroxide (H2O2) is both a source of oxidative damage and a signaling molecule. This protocol describes how to measure mitochondrial H2O2 using mitochondria-targeted HyPer7 (mtHyPer7), a genetically encoded ratiometric biosensor, in living yeast. It details how to optimize imaging conditions and perform quantitative cellular and subcellular analysis using freely available...

Source: Katsalifis, A., et al. Live Imaging of the Mitochondrial Glutathione Redox State in Primary Neurons using a Ratiometric Indicator. J. Vis. Exp. (2021)This video demonstrates the imaging of mitochondrial redox state in primary neurons using a ratiometric fluorescent indicator. By applying two excitation wavelengths under a fluorescence microscope, the fluorescence intensity ratio is measured to assess mitochondrial oxidation. The protocol includes N-methyl-D-aspartate (NMDA) stimulation...

This article describes a protocol to determine differences in basal redox state and redox responses to acute perturbations in primary hippocampal and cortical neurons using confocal live microscopy. The protocol can be applied to other cell types and microscopes with minimal...

We present step-by-step instructions for the generation of neonatal chimeras as well as the dissection and preparation of the thymus for ex vivo imaging by 2-Photon...

This study presents a simple, user-friendly ratiometric FRET assay for the detection and quantitative assessment of DNase activity, and demonstrates its application in the analysis of a weak...

A pH-sensitive ratiometric dye is used in combination with confocal laser scanning microscopy and digital image analysis to monitor extracellular pH in dental biofilms in...