Membrane tension
pores determine organelle dynamics and functions,
giving rise to physical observables during the cell death process.
While fluorescent organelle-targeted probes for specific chemical
analytes are increasingly available, subcellular dynamic processes
involving not only chemical parameters but also physicochemical and
physical parameters are uncommon. Here, we report a mitochondrial
chemical probe, named RCN, rationally designed to monitor
osmotic effects during transmembrane tension pore formation by using
local mitochondrial polarity and a subcellular localization redistribution
property of the probe. Utilizing fluorescence spectroscopy, high-resolution
confocal imaging, and spectrally resolved confocal microscopy, we
provide a new correlation between mitochondrial dynamics and bleb
vesicle formation using osmotic pressure stimuli in the cell, where
the mitochondrial local polarity was found to drastically increase.
The RCN provides a reliable protocol to assess transmembrane
pore formation driven by osmotic pressure increments through local
polarity variations and is a more robust physicochemical parameter
allowing the health and decease status of the cell to be measured.