Biosensing and monitoring of cell populations using the hydrogel bacterial microchip
Biosensing and monitoring of cell populations using the hydrogel bacterial microchip
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DOI：
10.1016/j.bios.2004.06.005
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发表时间：
2005-03-15
影响因子：
12.6
通讯作者：
Mirzabekov, AD
中科院分区：
文献类型：
--
作者：
Fesenko, DO;Nasedkina, TV;Mirzabekov, AD
提出了水凝胶细菌芯片技术的发展方向。该芯片是一组体积为0.3-60nl的半球形凝胶元件，附着在疏水玻璃表面，含有固定的活微生物细胞。单独的凝胶元件每个包含10(5)个细胞，即使在细胞分裂时也将它们保留在里面。凝胶的多孔结构使营养物质和被测物质易于进入固定化细胞。来自细胞的光信号很容易测量，并允许使用重要的荧光染色或编码生物发光或荧光报告的工程结构来监测细胞内代谢。研究了HBMChip的两种可能的应用模式，即细菌观察和生物传感。利用荧光染色法SYTO - 9分析了大肠杆菌生长过程中核酸合成的动态变化。有人建议用一个特殊的函数来评价细胞生长参数。HBMChip的生物传感性能已通过抗生素的定量分析和亚砷酸钠的检测得到验证。(C) 2004 Elsevier B.V.版权所有
Advanced development of the hydrogel bacterial microchip (HBMChip) technique is proposed. The microchip represents an array of hemispherical gel elements 0.3-60nl in volume attached to hydrophobic glass surface and containing live immobilized microbial cells. Separate gel elements contain each up to 10(5) cells and retain them inside even while the cells are dividing. Porous structure of the gel provides easy access of nutrients and tested substances to the immobilized cells. Optical signals from the cells are easily measurable and allow monitoring of intracellular metabolism using vital fluorescent stains or engineered constructs encoding bioluminescent or fluorescent reporters. Two possible application modes of the HBMChip have been investigated, i.e. the observation of bacteria and biosensing. The dynamics of nucleic acids synthesis in growing E. coli cells has been analyzed using vital fluorescent stain SYTO 9. A special function has been suggested for evaluation of the cell growth parameters. Biosensing properties of the HBMChip have been illustrated by quantitative analysis of antibiotics and the detection of sodium meta-arsenite. (C) 2004 Elsevier B.V. All rights reserved.