Digital concentration readout of single enzyme molecules using femtoliter arrays and Poisson statistics
Digital concentration readout of single enzyme molecules using femtoliter arrays and Poisson statistics
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DOI：
10.1021/nl060227d
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发表时间：
2006-03-01
期刊：
影响因子：
10.8
通讯作者：
Walt, DR
中科院分区：
文献类型：
--
作者：
Rissin, DM;Walt, DR
用于准确定量溶液中特定分析物浓度的方法都是基于集合响应，其中许多分析物分子产生测量信号。在这篇论文中，β-半乳糖苷酶的单分子是使用直径为1毫米的光纤束与2.4 × 10（5）单独密封，毫微微升微孔反应器进行监测。通过观察来自单个酶分子催化的荧光产物在反应容器阵列上的积累，并通过应用泊松统计分析，获得数字浓度读数。这种方法应该证明是有用的单分子酶学和超灵敏的生物测定。更一般地说，通过计数单个分子来确定浓度的能力提供了一种分析稀溶液的新方法。
Methods for accurately quantifying the concentration of a particular analyte in solution are all based on ensemble responses in which many analyte molecules give rise to the measured signal. In this paper, single molecules of beta-galactosidase were monitored using a 1 mm diameter fiber optic bundle with 2.4 x 10(5) individually sealed, femtoliter microwell reactors. By observation of the buildup of fluorescent products from single enzyme molecule catalysis over the array of reaction vessels and by application of a Poisson statistical analysis, a digital concentration readout was obtained. This approach should prove useful for single molecule enzymology and ultrasensitive bioassays. More generally, the ability to determine concentration by counting individual molecules offers a new approach to analysis of dilute solutions.