The production of heat by mitochondria is critical for maintaining body temperature, regulating metabolic rate, and preventing oxidative damage to mitochondria and cells. Until the present, mitochondrial heat production has been characterized only by methods based on fluorescent probes, which are sensitive to environmental variations (viscosity, pH, ionic strength, quenching, etc.). Here, for the first time, the heat release of isolated mitochondria was unambiguously measured by a diamond thermometer (DT), which is absolutely indifferent to external non-thermal parameters. We show that during total uncoupling of transmembrane potential by CCCP application, the temperature near the mitochondria rises by 4–22 °C above the ambient temperature with an absolute maximum of 45 °C. Such a broad variation in the temperature response is associated with the heterogeneity of the mitochondria themselves as well as their aggregations in the isolated suspension. Spontaneous temperature bursts with comparable amplitude were also detected prior to CCCP application, which may reflect involvement of some mitochondria to ATP synthesis or membrane potential leaking to avoid hyperproduction of reactive oxygen species. The results obtained with the diamond temperature sensor shed light on the “hot mitochondria” paradox.

We report a new approach to controllable thermal stimulation of a single living cell and its compartments. The technique is based on the use of a single polycrystalline diamond particle containing silicon-vacancy (SiV) color centers. Due to the presence of amorphous carbon at its intercrystalline boundaries, such a particle is an efficient light absorber and becomes a local heat source when illuminated by a laser. Furthermore, the temperature of such a local heater is tracked by the spectral shift of the zero-phonon line of SiV centers. Thus, the diamond particle acts simultaneously as a heater and a thermometer. In the current work, we demonstrate the ability of such a Diamond Heater-Thermometer (DHT) to locally alter the temperature, one of the numerous parameters that play a decisive role for the living organisms at the nanoscale. In particular, we show that the local heating of 11–12 °C relative to the ambient temperature (22 °C) next to individual HeLa cells and neurons, isolated from the mouse hippocampus, leads to a change in the intracellular distribution of the concentration of free calcium ions. For individual HeLa cells, a long-term (about 30 s) increase in the integral intensity of Fluo-4 NW fluorescence by about three times is observed, which characterizes an increase in the [Ca²⁺]_cyt concentration of free calcium in the cytoplasm. Heating near mouse hippocampal neurons also caused a calcium surge—an increase in the intensity of Fluo-4 NW fluorescence by 30% and a duration of ~ 0.4 ms.

Temperature is a crucial regulator of the rate and direction of biochemical reactions and cell processes. The recent data indicating the presence of local thermal gradients associated with the sites of high-rate thermogenesis, on the one hand, demonstrate the possibility for the existence of “thermal signaling” in a cell and, on the other, are criticized on the basis of thermodynamic calculations and models. Here, we review the main thermometric techniques and sensors developed for the determination of temperature inside living cells and diverse intracellular compartments. A comparative analysis is conducted of the results obtained using these methods for the cytosol, nucleus, endo-/sarcoplasmic reticulum, and mitochondria, as well as their biological consistency. Special attention is given to the limitations, possible sources of errors and ambiguities of the sensor’s responses. The issue of biological temperature limits in cells and organelles is considered. It is concluded that the elaboration of experimental protocols for ultralocal temperature measurements that take into account both the characteristics of biological systems, as well as the properties and limitations of each type of sensor is of critical importance for the generation of reliable results and further progress in this field.

Fluorescence spectra of nanodiamonds synthesized at high pressure from adamantane and other organic compounds show narrow (≈1 nm) lines of unknown origin over the spectroscopic range from ≈500 to 800 nm. The study proposes and experimentally confirms the hypothesis that these lines are related to radiative recombination of donor–acceptor pairs (DAP). According to the experimental data, these pairs can be formed from donor‐like substitutional nitrogen present in the diamond lattice and 2D acceptor layer resulting from the effect of transfer doping on the nanodiamond surface. A peculiar behavior of the narrow lines is identified within the temperature range of 100–10 K: their energy position slightly shifts downward, and the majority of the lines divide into two or more components as the temperature decreases. The lines are shown to be predominantly associated with single photon emitters, with an emission rate exceeding 1 million counts s ⁻¹ at room temperature. A new narrowband source of room‐temperature fluorescence found in hydrogen‐terminated nanodiamonds push horizons for quantum optical technologies related to the development of single photon emitters and temperature nanosensors.

Silicon-vacancy (SiV) color centers in diamond offer a promising system for quantum information applications owing to their intense narrowband emission and optically detectable spin states. Here, the fluorescent properties of ensembles and single SiV centers in diamonds obtained by high-pressure high-temperature synthesis from adamantane (below referred to as adamantane diamonds) are investigated at liquid-helium temperatures. Ensembles of SiV centers (~10³) are studied in large diamond crystals (1–2 µm in size). Despite the large number of excited centers, the fine structure of the zero-phonon line, corresponding to four allowed optical transitions between the ground- and excited-state doublets of the SiV center, can be observed in the fluorescence spectra of these crystals. The width of individual lines is in the range of 60–80 GHz, which is explained by their inhomogeneous broadening. Single SiV centers are studied in diamond crystallites with sizes of about 200 nm. The width of the narrowest transition lines observed in the fluorescence spectra of single SiV centers under resonant excitation is almost three orders of magnitude smaller than linewidths in the spectra of SiV ensembles and is as small as 94 MHz; i.e., these lines are so narrow that their width is determined by the lifetime of the upper state of the transition. Therefore, adamantane nanodiamond features the narrowest emission line width of a single SiV center at cryogenic temperatures among other types of SiV-con-taining nanodiamonds of similar sizes obtained by the high-pressure high-temperature and chemical vapor deposition techniques.

Despite progress in the high-pressure synthesis of nanodiamonds from hydrocarbons, the problem of controlled formation of fluorescent impurity centers in them still remains unresolved. In our work, we explore the potential of a new precursor composition, a mixture of adamantane with detonation nanodiamond, both in the synthesis of nanodiamonds and in the controlled formation of negatively charged silicon-vacancy centers in such nanodiamonds. Using different adamantane/detonation nanodiamond weight ratios, a series of samples was synthesized at a pressure of 7.5 GPa in the temperature range of 1200–1500 °C. It was found that temperature around 1350 °C, is optimal for the high-yield synthesis of nanodiamonds <50 nm in size. For the first time, controlled formation of negatively charged silicon-vacancy centers in such small nanodiamonds was demonstrated by varying the atomic ratios of silicon/carbon in the precursor in the range of 0.01–1%.