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Combined metal-enhanced fluorescence-surface acoustic wave (mef-saw) biosensor.

机译:结合金属增强的荧光表面声波(mef-saw)生物传感器。

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摘要

Immunofluorescence assays are capable of both detecting the amount of a protein and the location of the protein within a cell or tissue section. Unfortunately, the traditional technique is not capable of detecting concentrations on the nanoscale. Also, the technique suffers from non-specific attachment, which can cause false-positives, as well as photobleaching when detecting lower concentrations is attempted. There is also a time constraint problem since the technique can take from many hours to a few days in some cases. In this work, metal-enhanced fluorescence (MEF) is used to lower the detection limit and reduce photobleaching. Unfortunately, MEF also increases the intensity of non-specifically bound proteins (NSBPs). Therefore, a surface acoustic wave (SAW) device is used to remove the more weakly bound NSBPs. Previously, this has been shown on lithium niobate, but it is used with a quartz substrate in this work. The SAW device is also used to cause micro-mixing which speeds the process up significantly. In this research, it was found that silver nanocubes can lower the detection limit down to below 1 ng/mL. Quartz SAW devices are shown to remove NSBPs at a power of 10 mW applied for five minutes. Micro-mixing is shown to be improved by a factor of six at 10 mW for 10 minutes by saturating the antibody used in this research, which takes 1 hour without micro-mixing. Finally, all three components are combined. In this work, the whole device is used to detect 50 ng/mL. After micro-mixing, the intensity is the same as with MEF, and, after removal, it has been lowered by 7 a.u.
机译:免疫荧光测定法能够检测蛋白质的量以及蛋白质在细胞或组织切片中的位置。不幸的是,传统技术无法检测纳米级浓度。而且,该技术还具有非特异性附着的问题,这可能导致假阳性,并且在尝试检测较低浓度时会发生光漂白。还有一个时间约束问题,因为在某些情况下该技术可能要花费数小时到数天的时间。在这项工作中,金属增强荧光(MEF)用于降低检测限并减少光致漂白。不幸的是,MEF还增加了非特异性结合蛋白(NSBPs)的强度。因此,使用表面声波(SAW)装置来去除绑定较弱的NSBP。以前,这已在铌酸锂上显示出来,但在这项工作中它与石英衬底一起使用。 SAW设备还用于引起微混合,从而显着加快了处理速度。在这项研究中,发现银纳米立方体可以将检出限降低至1 ng / mL以下。 Quartz SAW设备显示在5分钟内以10 mW的功率去除了NSBP。通过使本研究中使用的抗体饱和,在10 mW的功率下进行10分钟的微混合可提高6倍,而无需微混合则需要1个小时。最后,将所有三个组件组合在一起。在这项工作中,整个设备用于检测50 ng / mL。微混合后,强度与MEF相同,去除后强度降低了7a.u。

著录项

  • 作者

    Morrill, Samuel.;

  • 作者单位

    University of South Florida.;

  • 授予单位 University of South Florida.;
  • 学科 Chemistry Biochemistry.;Health Sciences Immunology.;Engineering Biomedical.
  • 学位 M.Ch.E.
  • 年度 2014
  • 页码 86 p.
  • 总页数 86
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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