Sigaeva Alina, Hochstetter Axel, Bouyim Sighom, Chipaux Mayeul, Stejfova Miroslava, Cigler Petr, Schirhagl Romana
University Medical Center Groningen, Groningen University, Antonius Deusinglaan 1, Groningen, 9713AV, The Netherlands.
Research & Development, Life on a Chip e.K., Brunnenaecker 5, 73571, Goeggingen, Germany.
Small. 2022 Sep;18(39):e2201395. doi: 10.1002/smll.202201395. Epub 2022 Aug 29.
Diamond magnetometry can provide new insights on the production of free radicals inside live cells due to its high sensitivity and spatial resolution. However, the measurements often lack intracellular context for the recorded signal. In this paper, the possible use of single-particle tracking and trajectory analysis of fluorescent nanodiamonds (FNDs) to bridge that gap is explored. It starts with simulating a set of different possible scenarios of a particle's movement, reflecting different modes of motion, degrees of confinement, as well as shapes and sizes of that confinement. Then, the insights from the analysis of the simulated trajectories are applied to describe the movement of FNDs in glycerol solutions. It is shown that the measurements are in good agreement with the previously reported findings and that trajectory analysis yields meaningful results, when FNDs are tracked in a simple environment. Then the much more complex situation of FNDs moving inside a live cell is focused. The behavior of the particles after different incubation times is analyzed, and the possible intracellular localization of FNDs is deducted from their trajectories. Finally, this approach is combined with long-term magnetometry methods to obtain maps of the spin relaxation dynamics (or T1) in live cells, as FNDs move through the cytosol.
由于具有高灵敏度和空间分辨率，金刚石磁力测量法能够为活细胞内自由基的产生提供新的见解。然而，对于所记录的信号，这些测量往往缺乏细胞内的背景信息。在本文中，我们探索了利用荧光纳米金刚石（FNDs）的单粒子跟踪和轨迹分析来弥补这一差距的可能性。首先模拟了一组粒子运动的不同可能场景，反映了不同的运动模式、限制程度以及限制的形状和大小。然后，将模拟轨迹分析得出的见解应用于描述FNDs在甘油溶液中的运动。结果表明，在简单环境中跟踪FNDs时，测量结果与先前报道的发现高度吻合，并且轨迹分析产生了有意义的结果。接着重点研究了FNDs在活细胞内运动这一更为复杂的情况。分析了不同孵育时间后粒子的行为，并从其轨迹推断出FNDs可能的细胞内定位。最后，当FNDs在细胞质中移动时，将这种方法与长期磁力测量方法相结合，以获得活细胞中自旋弛豫动力学（或T1）的图谱。
