Prokhorov General Physics Institute of the Russian Academy of Sciences, Vavilov str. 38, Moscow, 119991, Russia.
Institute of Theoretical and Experimental Biophysics of the Russian Academy of Sciences, Pushchino, Moscow Region, 142292, Russia.
Sci Rep. 2023 May 26;13(1):8546. doi: 10.1038/s41598-023-35141-4.
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 [Ca] 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.
我们报告了一种新的方法,可以对单个活细胞及其隔室进行可控的热刺激。该技术基于使用单个含有硅空位 (SiV) 色心的多晶金刚石颗粒。由于其晶界存在非晶态碳,因此这种颗粒是一种高效的光吸收体,当被激光照射时,它会成为局部热源。此外,通过 SiV 中心零声子线的光谱位移来跟踪这种局部加热器的温度。因此,金刚石颗粒同时充当加热器和温度计。在当前的工作中,我们证明了这种金刚石加热-温度计(DHT)有能力局部改变温度,这是众多对纳米尺度下的生物体起决定性作用的参数之一。具体来说,我们表明,在 22°C 的环境温度下,在单个 HeLa 细胞和从小鼠海马体中分离的神经元旁边进行 11-12°C 的局部加热,会导致游离钙离子浓度的细胞内分布发生变化。对于单个 HeLa 细胞,观察到 Fluo-4 NW 荧光的积分强度约增加三倍,这表明细胞质中游离钙离子的[Ca]浓度增加。在小鼠海马体神经元附近加热也会引起钙涌——Fluo-4 NW 荧光强度增加 30%,持续时间约为 0.4ms。