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低成本的钙荧光法用于活细胞中纳米颗粒的长期研究。

Low-cost calcium fluorometry for long-term nanoparticle studies in living cells.

机构信息

Department of Electrical and Computer Engineering, Montana State University, Bozeman, Montana, 59717, USA.

Department of Physics, North Carolina State University, Raleigh, NC, 27695, USA.

出版信息

Sci Rep. 2020 Jul 28;10(1):12568. doi: 10.1038/s41598-020-69412-1.

Abstract

Calcium fluorometry is critical to determine cell homeostasis or to reveal communication patterns in neuronal networks. Recently, characterizing calcium signalling in neurons related to interactions with nanomaterials has become of interest due to its therapeutic potential. However, imaging of neuronal cell activity under stable physiological conditions can be either very expensive or limited in its long-term capability. Here, we present a low-cost, portable imaging system for long-term, fast-scale calcium fluorometry in neurons. Using the imaging system, we revealed temperature-dependent changes in long-term calcium signalling in kidney cells and primary cortical neurons. Furthermore, we introduce fast-scale monitoring of synchronous calcium activity in neuronal cultures in response to nanomaterials. Through graph network analysis, we found that calcium dynamics in neurons are temperature-dependent when exposed to chitosan-coated nanoparticles. These results give new insights into nanomaterial-interaction in living cultures and tissues based on calcium fluorometry and graph network analysis.

摘要

钙荧光法对于确定细胞内稳态或揭示神经元网络中的通讯模式至关重要。由于其治疗潜力,最近,研究与纳米材料相互作用的神经元中的钙信号成为了关注点。然而,在稳定的生理条件下对神经元细胞活性进行成像,要么非常昂贵,要么在长期能力方面受到限制。在这里,我们提出了一种低成本、便携式的成像系统,用于在神经元中进行长期、快速的钙荧光法测量。使用该成像系统,我们揭示了在肾脏细胞和原代皮质神经元中,长期钙信号随温度的变化。此外,我们引入了对纳米材料刺激下神经元培养物中同步钙活性的快速尺度监测。通过图网络分析,我们发现壳聚糖包覆的纳米颗粒暴露时,神经元中的钙动力学随温度变化。这些结果基于钙荧光法和图网络分析,为活细胞和组织中纳米材料相互作用提供了新的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9b26/7387557/885dbc8de056/41598_2020_69412_Fig1_HTML.jpg

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