Bio-Cell Gene Expression Analyzer

术语“生物细胞”指的是在胁迫作用的整个实验过程中保持其存活状态的细胞。这项研究是为了演示生物细胞对各种应激信号的反应的实时分析。电应力信号被施加到烟草或水稻的培养细胞上。作为细胞反应，用显微荧光图像分析系统测量基因表达。通过烟草细胞株BY-2的实验，测量了不同强度的脉冲电应力信号下细胞膜的电阻抗。结果表明，当跨膜电位(V&lt；Cp&gt；)超过200 mV时，膜有明显的通透性。另一方面，利用水稻原生质体进行的两步实验证明，几丁质酶基因的表达依赖于钙离子。在第一步，由脉冲电应力信号控制钙离子的引入。结果表明，在VCMP的1-5 mV范围内，Ca~(2+)&Gt~(2+)进入细胞内。作为第二步，我们将Ca~(2+)&Gt~(2+)直接注入原生质体中，证实了几丁质酶基因的表达。除了电应力，我们还打算在生物细胞内部施加电化学应力。然后，我们研制了直径为1-2μm的毛细碳微电极，并通过分析多巴胺的循环伏安图来检测其性能。综上所述，开发了生物细胞基因表达分析系统，并展示了其良好的性能。

The term "bio-cell" means a cell that maintains its viable state throughout the experiments of stress application. This study was done in order to demonstrate the real time analysis of the bio-cell responses to various stress signals. An electric stress signal was applied to cultured cells of tobacco or rice. As the cell response, the gene expression was measured with a microscopic fluorescent image analyzing system. By the experiment using tobacco cells, line BY-2, the electric impedance of the cell membrane was measured under various intensities of pulsing electric stress signals. As the result, appreciable membrane permeabilization was observed when the cross membrane potential (V_<CMP>) exceeded 200mV.On the other hand, the Ca^<2+>-dependent expression of chitinase gene was demonstrated by 2-step experiments using rice protoplasts. At the first step, Ca^<2+> introduction was controlled by the pulsing electric stress signal. As the result, Ca^<2+> entered the cells around 1-5 mV of VCMP.As the 2nd step, we demonstrated the chitinase gene expression by the direct injection of Ca^<2+> into the protoplasts. Besides the electric stress, we intended to apply an electrochemical stress to a bio-cell at its inside. Then we developed a capillary carbon microelectrode with a 1-2 μm diameter and checked its performance by analyzing cyclicvoltammograms of dopamine. In conclusion, the analyzing system of the gene expression of a bio-cell was developed and its promising performance was demonstrated.