Ultrasensitive and high-throughput fluorescence analysis of droplet contents with orthogonal line confocal excitation.
Ultrasensitive and high-throughput fluorescence analysis of droplet contents with orthogonal line confocal excitation.
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DOI：
10.1021/ac102173m
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发表时间：
2010-12-01
影响因子：
7.4
通讯作者：
Chiu, Daniel T.
中科院分区：
文献类型：
--
作者：
Jeffries, Gavin D. M.;Lorenz, Robert M.;Chiu, Daniel T.
本文描述了一种对传统共聚焦荧光检测的简单改进，大大提高了液滴流高速分析的信噪比（S/N）。而不是使用传统的外射几何形状，液滴的照明是在一条线的形式，这是正交的流动方向和光收集物镜。与我们从液滴观察到高水平散射背景的epi几何相反，当使用正交线共聚焦照明时，我们检测到的背景（取决于所使用的激光功率）减少了10倍以上。我们在分析物浓度范围内使用标准微流控平台表征了这种改进，并观察到检测限大于10的改进。使用这种方法，我们能够以大于350滴/秒的速度分析皮升体积液滴中含有的分析物的皮摩尔浓度。
This paper describes a simple modification to traditional confocal fluorescence detection that greatly improves signal-to-noise (S/N) for the high-speed analysis of droplet streams. Rather than using the conventional epi geometry, illumination of the droplet was in the form of a line that is orthogonal to both the direction of flow and the light-collection objective. In contrast to the epi geometry where we observed high levels of scattering background from the droplets, we detected more than 10-fold less background (depending on the laser power used) when orthogonal-line-confocal illumination was used. We characterized this improvement using a standard microfluidic platform over a range of analyte concentrations, and observed an improvement in limits of detection of greater than 10. Using this method, we were able to analyze pico-molar concentrations of analytes contained within picoliter-volume droplets at a rate of greater than 350 droplets per second.
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影响因子：
7.4
作者：
Roman, Gregory T.;Wang, Meng;Shultz, Kristin N.;Jennings, Colin;Kennedy, Robert T.
通讯作者：
Kennedy, Robert T.
影响因子：
7.4
作者：
Chiu DT;Lorenz RM;Jeffries GD
通讯作者：
Jeffries GD
影响因子：
14.9
作者：
Chen C;Ridzon DA;Broomer AJ;Zhou Z;Lee DH;Nguyen JT;Barbisin M;Xu NL;Mahuvakar VR;Andersen MR;Lao KQ;Livak KJ;Guegler KJ
通讯作者：
Guegler KJ
影响因子：
7.4
作者：
He, MY;Sun, CH;Chiu, DT
通讯作者：
Chiu, DT
影响因子：
6.2
作者：
Lorenz, Robert M.;Fiorini, Gina S.;Jeffries, Gavin D. M.;Lim, David S. W.;He, Mingyan;Chiu, Daniel T.
通讯作者：
Chiu, Daniel T.