Single-cell-based sensors and synchrotron FTIR spectroscopy: a hybrid system towards bacterial detection.
Single-cell-based sensors and synchrotron FTIR spectroscopy: a hybrid system towards bacterial detection.
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DOI：
10.1016/j.bios.2007.04.010
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发表时间：
2007-09
影响因子：
12.6
通讯作者：
Mandana Veiseh;Omid Veiseh;Michael C. Martin;C. Bertozzi;Miqin Zhang
中科院分区：
文献类型：
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作者：
Mandana Veiseh;Omid Veiseh;Michael C. Martin;C. Bertozzi;Miqin Zhang
单巨噬细胞为基础的传感器的微阵列的开发和证明潜在的实时细菌检测同步辐射FTIR显微镜。通过表面工程技术将细胞图案化在氧化硅基底的金电极上，其中金电极用纤连蛋白固定以介导细胞粘附，并且氧化硅背景用聚乙二醇（PEG）钝化以抵抗蛋白质吸附和细胞粘附。比较了暴露于不同浓度脂多糖（LPS）的金电极上的单细胞、双细胞和三细胞的细胞形态和红外光谱，以揭示这种基于细胞的传感平台的检测能力。发现基于单细胞的系统在暴露于LPS时产生最显著和一致的IR光谱偏移，从而提供最高的检测灵敏度。结果表明，细胞形态的变化和细胞暴露于LPS后的IR位移与LPS浓度和暴露时间有关，从而建立了一种鉴定LPS浓度和感染细胞群的方法。通过比较金电极表面积分别为25、100和400μ m2的单电池阵列的红外光谱，研究了将该单电池系统与常规红外光谱仪结合使用的可能性及其局限性。这种基于细胞的平台可能提供实时、无标记和快速的细菌检测，并允许高通量统计分析和便携性。
Microarrays of single macrophage cell-based sensors were developed and demonstrated for potential real-time bacterium detection by synchrotron FTIR microscopy. The cells were patterned on gold electrodes of silicon oxide substrates by a surface engineering technique, in which the gold electrodes were immobilized with fibronectin to mediate cell adhesion and the silicon oxide background was passivated with polyethylene glycol (PEG) to resist protein adsorption and cell adhesion. Cell morphology and IR spectra of single, double, and triple cells on gold electrodes exposed to lipopolysaccharide (LPS) of different concentrations were compared to reveal the detection capability of this cell-based sensing platform. The single-cell-based system was found to generate the most significant and consistent IR spectrum shifts upon exposure to LPS, thus providing the highest detection sensitivity. Changes in cell morphology and IR shifts upon cell exposure to LPS were found to be dependent on the LPS concentration and exposure time, which established a method for the identification of LPS concentration and infected cell population. Possibility of using this single-cell system with conventional IR spectroscopy as well as its limitation was investigated by comparing IR spectra of single-cell arrays with gold electrode surface areas of 25, 100, and 400μm2using both synchrotron and conventional FTIR spectromicroscopes. This cell-based platform may potentially provide real-time, label-free, and rapid bacterial detection, and allow for high-throughput statistical analyses, and portability.