Key Laboratory of Photochemical Conversion and Optoelectronic Materials, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, Beijing 100190, China.
University of Chinese Academy of Sciences, Beijing 100049, China.
ACS Chem Neurosci. 2020 May 6;11(9):1283-1290. doi: 10.1021/acschemneuro.0c00041. Epub 2020 Apr 28.
Ionic calcium (Ca) is an important second messenger in cells, particularly in the neuron. A deficiency or excess of Ca would lead to neuronal apoptosis and further injury to the brain. For accurate analysis of intracellular Ca, a single silicon nanowire (SiNW)-based ratiometric biosensor was constructed by simultaneously anchoring Ru(bpy)(mcbpy-O-Su-ester)(PF), as a reference molecule, and Fluo-3, as a response molecule, onto the surface of a single SiNW. The SiNW-based biosensor exhibits high sensitivity and favorable selectivity for detecting Ca. With the assistance of a micromanipulator and laser scanning confocal microscope, two single SiNW sensors were placed in the body and the neurites of an individual neuron to detect Ca. The difference between the concentrations of Ca in the body and neurites was identified. The results from the present study provide new insights into Ca in neurons at a high spatial resolution, and the strategy used in this study provides a new opportunity to investigate cellular metabolism by combining the advantages of a single-cell detection technique and physiology.
离子钙（Ca）是细胞中一种重要的第二信使，特别是在神经元中。Ca 的缺乏或过量会导致神经元凋亡，并进一步损伤大脑。为了准确分析细胞内的 Ca，通过将 Ru(bpy)(mcbpy-O-Su-ester)(PF)（作为参考分子）和 Fluo-3（作为响应分子）同时固定在单个硅纳米线（SiNW）的表面上，构建了一个基于单个 SiNW 的比率型生物传感器。基于 SiNW 的生物传感器对 Ca 的检测具有高灵敏度和良好的选择性。在微操纵器和激光扫描共聚焦显微镜的协助下，将两个单个 SiNW 传感器放置在单个神经元的体内和突起中以检测 Ca。确定了体内和突起中 Ca 浓度的差异。本研究的结果以高空间分辨率提供了对神经元中 Ca 的新见解，并且本研究中使用的策略通过结合单细胞检测技术和生理学的优势，为研究细胞代谢提供了新的机会。
