Rondelez Yannick, Tresset Guillaume, Tabata Kazuhito V, Arata Hideyuki, Fujita Hiroyuki, Takeuchi Shoji, Noji Hiroyuki
Nat Biotechnol. 2005 Mar;23(3):361-5. doi: 10.1038/nbt1072. Epub 2005 Feb 20.
Precise understanding of biological functions requires tools comparable in size to the basic components of life. Single molecule studies have revealed molecular behaviors usually hidden in the ensemble- and time-averaging of bulk experiments. Although most such approaches rely on sophisticated optical strategies to limit the detection volume, another attractive approach is to perform the assay inside very small containers. 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.
对生物功能的精确理解需要尺寸与生命基本组成部分相当的工具。单分子研究揭示了通常隐藏在大量实验的总体平均和时间平均中的分子行为。尽管大多数此类方法依赖复杂的光学策略来限制检测体积，但另一种有吸引力的方法是在非常小的容器内进行分析。我们开发了一种带有大量微米级腔体的硅树脂装置。我们用它长时间紧密封装低至飞升至的溶液体积。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单分子的活性证明了我们方法的可行性。该方法对于单分子水平的许多超灵敏生物测定应该是有意义的。
