Glucose and lactate biosensors for scanning electrochemical microscopy imaging of single live cells
Glucose and lactate biosensors for scanning electrochemical microscopy imaging of single live cells
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DOI：
10.1021/ac7021184
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发表时间：
2008-04-15
影响因子：
7.4
通讯作者：
Cliffel, David E.
中科院分区：
文献类型：
--
作者：
Ciobanu, Madalina;Taylor, Dale E., Jr.;Cliffel, David E.
我们开发了基于葡萄糖氧化酶和乳酸氧化酶的葡萄糖和乳酸超微电极（UME）生物传感器（通过电聚合或铸造将酶固定在Pt上）用于扫描电化学显微镜（SECM）并分别测定了它们对葡萄糖和乳酸的敏感性。我们的评估结果揭示了每种方法构建的传感器的不同优势：提高了灵敏度，缩短了手工铸造的制造时间，提高了电聚合的可重复性。我们已经获得了每种制造的生物传感器UME的安培接近曲线（ac），这些ac可以用作将UME定位在已知距离上的基板上的手段。我们使用葡萄糖生物传感器来记录单个成纤维细胞以上的葡萄糖摄取情况。同样，我们也使用了乳酸生物传感器me，用SECM记录单个癌细胞以上的乳酸产量。我们还表明，单个癌细胞的氧呼吸谱并不模拟细胞地形，而是相当复杂，在细胞接触培养皿的地方观察到更高的呼吸活性。这些UME生物传感器，连同其他已经在文献中描述的应用，可能被证明是在单个癌细胞中绘制代谢分析物（如葡萄糖、乳酸和氧）的强大工具。
We have developed glucose and lactate ultramicroelectrode (UME) biosensors based on glucose oxidase and lactate oxidase (with enzymes immobilized onto Pt UMEs by either electropolymerization or casting) for scanning electrochemical microscopy (SECM) and have determined their sensitivity to glucose and lactate, respectively. The results of our evaluations reveal different advantages for sensors constructed by each method: improved sensitivity and shorter manufacturing time for hand-casting, and increased reproducibility for electropolymerization. We have acquired amperometric approach curves (ACs) for each type of manufactured biosensor UME, and these ACs can be used as a means of positioning the UME above a substrate at a known distance. We have used the glucose biosensor UMEs to record profiles of glucose uptake above individual fibroblasts. Likewise, we have employed the lactate biosensor UMEs for recording the lactate production above single cancer cells with the SECM. We also show that oxygen respiration profiles for single cancer cells do not mimic cell topography, but are rather more convoluted, with a higher respiration activity observed at the points where the cell touches the Petri dish. These UME biosensors, along with the application of others already described in the literature, could prove to be powerful tools for mapping metabolic analytes, such as glucose, lactate, and oxygen, in single cancer cells.