Microfabricated arrays of femtoliter chambers allow single molecule enzymology
Microfabricated arrays of femtoliter chambers allow single molecule enzymology
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DOI：
10.1038/nbt1072
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发表时间：
2005-03-01
影响因子：
46.9
通讯作者：
Noji, H
中科院分区：
文献类型：
--
作者：
Rondelez, Y;Tresset, G;Noji, H
精确理解生物功能需要与生命基本组成部分大小相当的工具（1-4）。单分子研究揭示了通常隐藏在整体实验和时间平均中的分子行为（5，6）。尽管大多数这样的方法依赖于复杂的光学策略来限制检测体积（7，8），但另一种有吸引力的方法是在非常小的容器内进行测定（9-16）。我们已经开发了一种硅胶装置，其呈现出大量微米尺寸的腔体。我们用它来紧密地封闭溶液体积，低至毫微微升，在很长一段时间。微芯片确保腔室均匀且精确定位。我们通过测量β-半乳糖苷酶和辣根过氧化物酶的单分子活性证明了我们方法的可行性。该方法应该是在单分子水平上的许多超灵敏的生物测定的兴趣。
Precise understanding of biological functions requires tools comparable in size to the basic components of life(1-4). Single molecule studies have revealed molecular behaviors usually hidden in the ensemble- and time-averaging of bulk experiments(5,6). Although most such approaches rely on sophisticated optical strategies to limit the detection volume(7,8), another attractive approach is to perform the assay inside very small containers(9-16). We have developed a silicone device presenting a large array of micrometer-sized cavities. We used it to tightly enclose volumes of solution, as low as femtoliters, over long periods of time. The microchip insures that the chambers are uniform and precisely positioned. We demonstrated the feasibility of our approach by measuring the activity of single molecules of beta-galactosidase and horseradish peroxidase. The approach should be of interest for many ultrasensitive bioassays at the single-molecule level.