School of Basic Sciences, Institute of Physics, EPFL, Rte Cantonale, 1015, Lausanne, Switzerland.
Division of Quantum Metrology and Nanotechnologies, Istituto Nazionale di Ricerca Metrologica (INRiM), Strada delle Cacce 91, 10135, Torino, Italy.
Sci Rep. 2023 Apr 11;13(1):5909. doi: 10.1038/s41598-023-32235-x.
Monitoring neuronal activity with simultaneously high spatial and temporal resolution in living cell cultures is crucial to advance understanding of the development and functioning of our brain, and to gain further insights in the origin of brain disorders. While it has been demonstrated that the quantum sensing capabilities of nitrogen-vacancy (NV) centers in diamond allow real time detection of action potentials from large neurons in marine invertebrates, quantum monitoring of mammalian neurons (presenting much smaller dimensions and thus producing much lower signal and requiring higher spatial resolution) has hitherto remained elusive. In this context, diamond nanostructuring can offer the opportunity to boost the diamond platform sensitivity to the required level. However, a comprehensive analysis of the impact of a nanostructured diamond surface on the neuronal viability and growth was lacking. Here, we pattern a single crystal diamond surface with large-scale nanopillar arrays and we successfully demonstrate growth of a network of living and functional primary mouse hippocampal neurons on it. Our study on geometrical parameters reveals preferential growth along the nanopillar grid axes with excellent physical contact between cell membrane and nanopillar apex. Our results suggest that neuron growth can be tailored on diamond nanopillars to realize a nanophotonic quantum sensing platform for wide-field and label-free neuronal activity recording with sub-cellular resolution.
在活细胞培养物中以同时具有高空间和时间分辨率监测神经元活动对于深入了解大脑的发育和功能至关重要,并有助于进一步了解大脑疾病的起源。虽然已经证明,钻石中的氮空位(NV)中心的量子感应能力允许实时检测海洋无脊椎动物中大神经元的动作电位,但迄今为止,对哺乳动物神经元(呈现出更小的尺寸,因此产生的信号更低,需要更高的空间分辨率)的量子监测仍然难以实现。在这种情况下,钻石纳米结构化可以提供机会将钻石平台的灵敏度提高到所需的水平。然而,对于纳米结构化钻石表面对神经元活力和生长的影响缺乏全面分析。在这里,我们用大尺寸纳米柱阵列对单晶钻石表面进行图案化,并成功地证明了活的和功能正常的原代小鼠海马神经元网络在其表面上的生长。我们对几何参数的研究表明,细胞沿着纳米柱网格轴优先生长,细胞膜和纳米柱顶点之间具有极好的物理接触。我们的结果表明,可以根据神经元的生长来调整钻石纳米柱,以实现用于宽场和无标记神经元活动记录的纳米光子量子感应平台,具有亚细胞分辨率。