Development of Real-time Observation Method for Cell Response without Probes and Its Application to Biosensing

本研究开发了一种新型的基于细胞的高分辨率二维表面等离子体共振（2D-SPR）生物传感系统。利用二维表面等离子体共振成像仪实时观察了粘附在Au芯片上的神经细胞或内分泌细胞对激动剂刺激的反应。进一步证明了2D-SPR成像中对激动剂刺激的细胞反应可能是基于信号转导蛋白如PKC向细胞膜的细胞内募集。我们已经发现，细胞，如圆柱状细胞，如分裂酵母，在高频交流电场下的电取向明显依赖于细胞的活力。这些结果表明，无标记细胞，基于生物传感的2D-SPR成像方法或电定向观察方法应用于药物筛选、细胞状态评估、细胞识别等。

In this research, novel cell-based biosensing system with a high resolution of two dimensional surface plasmon resonance (2D-SPR) imager was developed. Response of some neural or endocrine cells which were adhered on an Au chip to agonist stimulation was successfully observed by the reflection intensity change in the individual cell regions with the 2D-SPR imager at real-time. It was further demonstrated that the cell response in 2D-SPR imaging to agonist stimulation might be based on intracellular recruitment of signal transduction proteins such as PKC to cell membrane.Furthermore, we have found that the electro-orientation of cells such as cylinder-like cells such as fission yeast under high frequency AC electric field clearly depended on the cell viability.These results suggested future wide applications of label-free cell-based biosensing with 2D-SPR imaging method or electro-orientation observation method to drug screening, cell state evaluation, cell recognition and so on.