Fabrication and Characterization of Biomolecule-based Structure by Using SECM

1.扫描电化学显微镜（SECM）用于局部电产生羟基自由基，以在具有不同烷基硅烷衍生物的自组装单层（SAMs）的玻璃基底上产生具有心肌黄酶的图案。SECM探针（微电极）电生成的羟基自由基扩散到SAM表面，降解SAM并将其从玻璃表面去除。当SAM是疏水性的，心肌黄酶模式形成的基板上，通过浸渍所得的基板到心肌黄酶溶液。还使用具有功能性末端基团的SAM通过化学键合来修饰黄递酶，在底物上给出黄递酶模式。SECM用于在单细胞水平上对细胞活性成像。基于氧气还原电流的SW-480的SECM测量给出了呼吸活动的图像。活细胞的SECM测量特别适合于阐明化学和物理刺激对细胞的影响。 关于我们 在单个细胞水平上的定位。当在培养基中加入氰根离子或硝基酚时，细胞的SECM图像变得苍白，表明这些物质降低了呼吸活性。通过定量分析细胞活性的变化来评价细胞膜对氰离子的渗透。同样，SECM也被用于估算单个植物细胞的呼吸和光合活性. SECM用于估计几种氧化还原物质通过单个藻类原生质体的细胞膜的渗透。当微电极靠近影响半球形扩散区形成的膜放置时，氧化还原电流变得低于在本体溶液中观察到的电流。由于氧化还原电流的降低与膜对氧化还原物质的渗透性相关，因此对该现象的定量分析提供了氧化还原物质穿过膜的渗透系数（Pm）。Fe（CN）_6 ~（2-）<-4>、Fe（CN）_6 ~（3-）、Co（phen）_3 ~（2-）、FMA和氢醌的Pm值分别为&lt;1 × 10 ~（-4）、&lt;1 × 10 ~（-4）、1 × 10 ~（-4）、<-3>5 × 10 ~<-3>（-4）和2 × 10 ~（-4<-2>）cm/sec。少

1. Scanning electrochemical microscopy (SECM) was used for localized electrogeneration of hydroxyl radical to create patterns with diaphorase on glass substrates with self-assembled mono-layers (SAMs) of different alkylsilane derivatives. The electrogenerated hydroxyl radical from the probe (microelectrode) of SECM diffused onto the SAM surfaces and degraded the SAMs and removed them from the glass surfaces. When the SAM was hydrophobic, diaphorase patterns were formed on the substrates by dipping the resulting substrate into a diaphorase solution. A SAM with functional terminal groups was also used to immobilize diaphorase via chemical linkage, giving a diaphorase pattern on the substrate.2. SECM was used for imaging of cellular activity at the single cell level. SECM measurements of SW-480 based on the reduction current of oxygen gave images of respiration activity. The SECM measurements of living cells are particularly suitable to clarify the influence of chemical and physical stimu … More lation at a single cell level. When cyanide ion or nitrophenol was added to the culture medium, the SECM image of the cell became pale, indicating that these species lowers the respiration activity. The permeation of the cellular membrane to cyanide ion was evaluated by the quantitative analysis of the change in the cellular activity. Similarly, SECM was used for estimating the respiration and photosynthetic activity of a single plant cell.3. SECM was used for estimation of permeation of several redox species through a cell membrane of a single algal protoplast. When the microelectrode is placed close to a membrane which influences the formation of the semi-spherical diffusion region, the redox current becomes lower than that observed in the bulk solution. Since the decrease in the redox current correlates with the permeability of the membrane to redox species, the quantitative analysis of the phenomena affords the permeation coefficient (Pm) for redox species to pass through the membrane. The Pm values for Fe(CN)_6^<-4>, Fe(CN)63-, Co(phen)32_3^<+2>, FMA and hydroquinone were <1x10-4, <1x10-4, 1x10^<-3>, 5x10^<-3> and 2x10^<-2> cm/sec, respectively. Less