Rougier, J.-S.; Glushkov, E.; Guichard, S.; Kucera, J.; Zeeb, V.; Abriel, H.

A living cell is a nonequilibrium thermodynamic system where, nevertheless, a notion of local equilibria exists. This notion applies to all micro- and nanoscale aqueous volumes, each containing a large number of molecules. This allows one to define sets of local conditions, including thermodynamic ones; for instance, a defined temperature requires thermodynamic equilibrium by definition. Once such a condition is fulfilled, one can control local variables and their gradients to theoretically describe the thermodynamic state of living systems at the micro- and nanoscale. Performing ultralocal experimental manipulations has become possible thanks to the patch clamp technique to control the cell membrane potential, as well as fluorescent imaging to monitor molecular concentrations and their intracellular gradients. However, precise temperature gradient control at the micro-/nanoscale has yet lacked a reliable experimental realization in a living cell. Here, we present a new methodology, microscale control of a temperature gradient profile in aqueous media by a fully optical Diamond Heater-Thermometer in a plug-and-play configuration combined with the patch clamp technique. In particular, we demonstrate applications of the combined Diamond Heater-Thermometer-patch clamp approach for the fast and reproducible thermal modulation of ionic current from voltage-gated Nav1.5 sodium channels expressed in HEK293 cells and freshly isolated ventricular mouse cardiomyocytes. Such an approach of manipulating the ultra-local temperature down to the nanoscale has the potential to uncover previously inaccessible phenomena in various physiological intracellular processes related to the endogenous nanoscale heat sources, such as open ion channels capable of producing Joule heat.