Wu Chunsheng, Du Liping, Tian Yulan, Zhang Xi, Wang Ping
Institute of Medical Engineering, School of Basic Medical Sciences, Health Science Center, Xi'an Jiaotong University, Xi'an, 710061, China.
Department of Biomedical Engineering, Zhejiang University, Hangzhou, 310027, China.
Methods Mol Biol. 2017;1572:233-246. doi: 10.1007/978-1-4939-6911-1_16.
A light-addressable potentiometric sensor (LAPS), a silicon-based surface potential detector, is combined with bioengineered olfactory sensory neurons (OSN) for odorant detection. A LAPS chip is used as a transducer to monitor cell membrane potential changes. In addition, a focused movable laser with a diameter comparable to cell sizes is employed to select the desirable single cell for measurement under a microscope. Bioengineered OSNs are coupled to the LAPS surface and employed as sensing elements, which are prepared by the expression of an olfactory receptor of C. elegans, ODR-10, on the plasma membrane of rat primary OSNs via transient transfection. The responses of bioengineered OSNs to diacetyl, isoamyl acetate, and acetic acid are monitored by extracellular recording using the LAPS chip. Features of the recorded extracellular potential firings are analyzed in frequency and time domains. We have shown that bioengineered OSNs can generate specific response signals upon the stimulation of diacetyl, which is the natural ligand of ODR-10. Moreover, different concentrations of diacetyl can elicit different temporal firing patterns in bioengineered OSNs, which permits the concentration detection of specific odorant molecules in solution.
一种光寻址电位传感器(LAPS),一种基于硅的表面电位探测器,与生物工程嗅觉感觉神经元(OSN)相结合用于气味检测。LAPS芯片用作换能器来监测细胞膜电位变化。此外,使用直径与细胞大小相当的聚焦移动激光在显微镜下选择理想的单细胞进行测量。生物工程OSN与LAPS表面耦合并用作传感元件,其通过将秀丽隐杆线虫的嗅觉受体ODR-10通过瞬时转染表达在大鼠原代OSN的质膜上来制备。使用LAPS芯片通过细胞外记录监测生物工程OSN对双乙酰、乙酸异戊酯和乙酸的反应。在频域和时域分析记录的细胞外电位放电的特征。我们已经表明,生物工程OSN在双乙酰刺激下可以产生特定的反应信号,双乙酰是ODR-10的天然配体。此外,不同浓度的双乙酰可以在生物工程OSN中引发不同的时间放电模式,这允许检测溶液中特定气味分子的浓度。