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Thermogenetics appeared recently as an evolutionary advance in methods of optical stimulation of nerve cells and uses focused light to activate light-sensitive cation channels in the neuron membrane. Light activation and opening of light-sensitive channels, which can be expressed genetically in any type of neuron, induces neuron membrane depolarization and a resultant action potential. In contrast to classical methods of optogenetics, which use the visible spectrum, thermogenetics uses channels sensitive to warming. This provides the opportunity for additional activation of these channels not only using IR light, but also other methods of warming nervous tissue, such as ultrasound or microwave radiation. The penetrability of living tissue to stimulation with IR radiation is an order of magnitude greater than that for visible spectrum radiation, which allows thermogenetics methods to be used in vivo without invasive surgical methods to “clear the way” for optogenetic stimulation. On the other hand, the thermal nature of the stimulation imposes additional limits on use of the method, as heating must be sufficiently gentle so as not to induce heat shock at the cellular level, and the threshold of activation must be sufficiently high for channel opening not to occur spontaneously at normal physiological temperatures.

Optically detected electron spin resonance in fiber-coupled nitrogen–vacancy (NV) centers of diamond is used to demonstrate a fiber-optic quantum thermometry of individual thermogenetically activated neurons. Laser-induced temperature variations read out from single neurons with the NV-diamond fiber sensor are shown to strongly correlate with the fluorescence of calcium-ion sensors, serving as online indicators of the inward Ca2+ current across the cell membrane of neurons expressing transient receptor potential (TRP) cation channels. Local laser heating above the TRP-channel activation threshold is shown to reproducibly evoke robust action potentials, visualized by calcium-ion-sensor-aided fluorescence imaging and detected as prominent characteristic waveforms in the time-resolved response of fluorescence Ca2+ sensors.

Thermal activation of transient receptor potential (TRP) cation channels is one of the most striking examples of temperature-controlled processes in cell biology. As the evidence indicating the fundamental role of such processes in thermosensation builds at a fast pace, adequately accurate tools that would allow heat receptor logic behind thermosensation to be examined on a single-cell level are in great demand. Here, we demonstrate a specifically designed fiber-optic probe that enables thermal activation with simultaneous online thermometry of individual cells expressing genetically encoded TRP channels. This probe integrates a fiber-optic tract for the delivery of laser light with a two-wire microwave transmission line. A diamond microcrystal fixed on the fiber tip is heated by laser radiation transmitted through the fiber, providing a local heating of a cell culture, enabling a well-controlled TRP-assisted thermal activation of cells. Online local temperature measurements are performed by using the temperature-dependent frequency shift of optically detected magnetic resonance, induced by coupling the microwave field, delivered by the microwave transmission line, to nitrogen—vacancy centers in the diamond microcrystal. Activation of TRP channels is verified by using genetically encoded fluorescence indicators, visualizing an increase in the calcium flow through activated TRP channels.

Exposure to a microwave field is shown to enable thermogenetic activation of individual cells in a culture of cell expressing thermosensitive ion channels. Integration of a microwave transmission line with an optical fiber and a diamond quantum thermometer has been shown to allow thermogenetic single-cell activation to be combined with accurate local online temperature measurements based on an optical detection of electron spin resonance in nitrogen–vacancy centers in diamond.