Mitokyne: A Ratiometric Raman Probe for Mitochondrial pH

Mitochondrial pH (pH_mito) is intimately related to mitochondrial function, and aberrant values for pH_mito are linked to several disease states. We report the design, synthesis, and application of mitokyne 1-the first small molecule pH_mito sensor for stimulated Raman scattering (SRS) microscopy. This ratiometric probe can determine subtle changes in pH_mito in response to external stimuli and the inhibition of both the electron transport chain and ATP synthase with small molecule inhibitors. In addition, 1 was also used to monitor mitochondrial dynamics in a time-resolved manner with subcellular spatial resolution during mitophagy providing a powerful tool for dissecting the molecular and cell biology of this critical organelle.