Rapid, single-molecule assays in nano/micro-fluidic chips with arrays of closely spaced parallel channels fabricated by femtosecond laser machining.
Rapid, single-molecule assays in nano/micro-fluidic chips with arrays of closely spaced parallel channels fabricated by femtosecond laser machining.
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DOI：
10.3390/s140815400
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发表时间：
2014-08-20
期刊：
影响因子：
--
通讯作者：
Davis LM
中科院分区：
文献类型：
--
作者：
Canfield BK;King JK;Robinson WN;Hofmeister WH;Davis LM
具有成本效益的药物发现取决于在减少试剂需求的同时增加测定通量。为此，我们正在开发一种超灵敏的基于荧光的平台，该平台包含一个纳米/微流体芯片，该芯片具有一系列紧密间隔的通道，用于单分子测定的并行光学读出。在这里，我们描述了使用直接飞秒激光加工制造数百个紧密间隔的通道上的表面上的熔融石英基板。通过粘合到旋涂有聚（二甲基硅氧烷）薄膜的显微镜盖玻片上来密封通道。单分子检测实验使用定制的宽视场显微镜进行。通道阵列由线生成红色二极管激光器进行落射照明，从而在500微米的视场内产生仅几微米厚的线焦点。将荧光标记的生物分子的稀水溶液加载到装置中，并用电子倍增CCD相机检测荧光，允许每个微通道的采集速率高达7 kHz。基于实验参数的匹配数字滤波用于对检测到的荧光进行初始快速评估。更详细的分析是通过荧光相关光谱。模拟荧光数据与实验值吻合良好。
Cost-effective pharmaceutical drug discovery depends on increasing assay throughput while reducing reagent needs. To this end, we are developing an ultrasensitive, fluorescence-based platform that incorporates a nano/micro-fluidic chip with an array of closely spaced channels for parallelized optical readout of single-molecule assays. Here we describe the use of direct femtosecond laser machining to fabricate several hundred closely spaced channels on the surfaces of fused silica substrates. The channels are sealed by bonding to a microscope cover slip spin-coated with a thin film of poly(dimethylsiloxane). Single-molecule detection experiments are conducted using a custom-built, wide-field microscope. The array of channels is epi-illuminated by a line-generating red diode laser, resulting in a line focus just a few microns thick across a 500 micron field of view. A dilute aqueous solution of fluorescently labeled biomolecules is loaded into the device and fluorescence is detected with an electron-multiplying CCD camera, allowing acquisition rates up to 7 kHz for each microchannel. Matched digital filtering based on experimental parameters is used to perform an initial, rapid assessment of detected fluorescence. More detailed analysis is obtained through fluorescence correlation spectroscopy. Simulated fluorescence data is shown to agree well with experimental values.
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影响因子：
2.9
作者：
Okagbare, Paul I.;Soper, Steven Allan
通讯作者：
Soper, Steven Allan
影响因子：
7.4
作者：
Li, Y;Buch, JS;Lee, CS
通讯作者：
Lee, CS
影响因子：
6.1
作者：
Hatch, Andrew C.;Fisher, Jeffrey S.;Lee, Abraham P.
通讯作者：
Lee, Abraham P.
DOI：
10.1007/s005420100114
发表时间：
2002-01-01
期刊：
MICROSYSTEM TECHNOLOGIES
影响因子：
--
作者：
Gerlach, A;Knebel, G;Schaller, T
通讯作者：
Schaller, T
影响因子：
6.1
作者：
Mckenna, Brian K.;Selim, A. A.;Ehrlich, Daniel J.
通讯作者：
Ehrlich, Daniel J.