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Development of a micro fluorescence detector for m-TAS based on surface plasmon field enhancement

基于表面等离子体场增强的m-TAS微型荧光探测器的研制

基本信息

批准号：
18510110
负责人：
ISHIDA Akito
金额：
$ 1.54万
依托单位：
Kyoto Prefectural University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18510110/
关键词：
plasmon enhanced excitation micro total analysis system silver-coated nanocone array fluorescence immunoassay liauid phase silver coating nanoimprinting micro fluidics PDMS

项目摘要

The aim of this study is development of a high-performance fluorescence detection method based on surface plasmon enhanced excitation for integration into a micro total analysis system (μ-TAS). We employed a PMMA nanocone array (diameter, 250nm ; height, 750 nm ; pitch, 250 nm ; macroscopic dimension, 5×5mm^2) prepared by nanoimprinting as a substrate. The surface was treated with tannic acid and silver nitrate successively. After immobilization of Texas-Red on the surface, fluorescence enhancement property of the nanocone array was estimated by comparison between the fluorescence intensity of the same areas in the presence and absence of nanocone array upon far-field illumination using a fluorescence microscope. The silver-coated nanocone array showed the fluorescence intensity approximately 50 times larger than that of the flat surface. This result suggests that far-field illumination of the silver-coated nanocone array induces local plasmons and the strongly enhanced electromagnetic field effectively excites the fluorophores immobilized on the surface. Then we have demonstrated a practical application of this excitation method. Anti-IgG was immobilized on a silver coated nanocone arrays and the two sections of the surface were covered with PDMS microfluid channels, respectively. Fluorescence labeled IgG solution and BSA solution were independently introduced into the two channels and then, after careful washing, fluorescence microscopic imaging was carried out. The clear difference in the fluorescence intensity between the two channels demonstrates that the silver-coated nanocone is useful for fluorescence bioassays as a plasmon-enhanced substrate.

本研究的目的是建立一种基于表面等离子体增强激发的高效荧光检测方法，并将其集成到微总量分析系统（μ-TAS）中。我们采用纳米印迹技术制备的PMMA纳米锥阵列（直径250nm，高度750nm，节距250nm，宏观尺寸5×5mm^2）作为衬底。分别用单宁酸和硝酸银进行表面处理。将Texas-Red固定在纳米锥阵列表面后，利用荧光显微镜对相同区域进行远场光照，比较纳米锥阵列存在和不存在时的荧光强度，评估纳米锥阵列的荧光增强性能。镀银纳米锥阵列的荧光强度约为平面的50倍。结果表明，镀银纳米锥阵列的远场光照可诱发局部等离子体激元，强增强的电磁场可有效激发固定在表面的荧光团。然后给出了该激励方法的实际应用。将抗igg固定在镀银纳米锥阵列上，并在两部分表面分别覆盖PDMS微流体通道。分别将荧光标记的IgG溶液和牛血清白蛋白溶液分别引入两个通道中，仔细洗涤后进行荧光显微成像。两个通道之间荧光强度的明显差异表明，镀银纳米锥作为等离子体增强底物可用于荧光生物测定。