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Microfluidic Cytometry Using Surface-Enhanced Raman Scattering Biotags.

机译：使用表面增强拉曼散射生物标签的微流式细胞术。

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摘要

In this dissertation, we combine microfluidics and surface-enhanced Raman spectroscopy (SERS) in order to create a platform for identification of cells labeled with SERS biotags (SBTs) based their multiple surface biomarkers. Microfluidic systems allow us to transport cells to the detection region and control their advection with unprecedented control and precision. It also help us with faster processes using smaller sample quantities. In combination with a powerful chemical detection method such as SERS, a platform capable of multiplexed identification of cells can be developed.;SERS is a molecular identification method based on enhancement of molecules' vibrational spectra due to plasmonic resonances. Metallic nanostructures are needed to achieve unparalleled enhancements that render SERS capable of detection single molecules.;Here, we use a flow-focusing microfluidic device to transport cells and micron-sized particle to a SERS interrogation region and collect their signature in a cytometry fashion. Three labeling and data analysis schemes were developed:;• In the first platform, the concept of identification of SERS-labeled particles in a microfluidic channel was put to test. As a proof-of-concept, micron-sized beads labeled with different combinations of two tags was used and successfully identified by means of ratiometric analysis of SERS signals.;• In In the second system the two-tag platform was used in order to identify circulating prostate cancer cells in a background of normal prostate cells. In this system, one SBT was used as a universal tag and the second SBT was targeting a specific biomarker that is overexpressed in cancer cells. We managed to detect a population of 1% cancer cells in a mixture of cancer and normal cells.;Finally, a multiplexed SERS identification system was developed which enabled us to discriminate between four populations of microbeads that were labeled with multiple cocktails of four SBTs. This platforms can be used for various immunophenotyping application where cells with multiple surface biomarkers are targeted.
机译：本文结合微流控技术和表面增强拉曼光谱技术（SERS），为基于多种表面生物标志物的SERS生物标签（SBTs）标记的细胞的鉴定提供平台。微流体系统使我们能够将细胞运输到检测区域，并以前所未有的控制和精度控制细胞的对流。它还有助于我们使用较少的样品量来加快处理速度。结合强大的化学检测方法（例如SERS），可以开发一种能够对细胞进行多重鉴定的平台。; SERS是一种基于等离子体共振产生的分子振动光谱增强的分子鉴定方法。需要金属纳米结构来实现无与伦比的增强，从而使SERS能够检测单个分子。在此，我们使用流聚焦微流控设备将细胞和微米级颗粒运输到SERS询问区域，并以流式细胞仪收集其特征。开发了三种标记和数据分析方案：•在第一个平台中，对在微流体通道中鉴定SERS标记颗粒的概念进行了测试。作为概念验证，使用了标记有两个标签不同组合的微米大小的珠子，并通过对SERS信号进行比例分析成功地鉴定了该珠子。•在第二个系统中，使用了两个标签的平台来在正常前列腺细胞的背景下识别循环中的前列腺癌细胞。在该系统中，一个SBT用作通用标签，第二个SBT靶向在癌细胞中过表达的特定生物标记。我们设法在癌细胞和正常细胞的混合物中检测到了1％的癌细胞。最后，开发了一个多重SERS识别系统，使我们能够区分四个带有四个SBT混合物的微珠种群。该平台可用于具有多种表面生物标记的细胞靶向的各种免疫表型应用。

著录项

  • 作者

    Hoonejani, Mehran R.;

  • 作者单位

    University of California, Santa Barbara.;

  • 授予单位 University of California, Santa Barbara.;
  • 学科 Mechanical engineering.;Analytical chemistry.
  • 学位 Ph.D.
  • 年度 2015
  • 页码 95 p.
  • 总页数 95
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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