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首页> 外文学位 >Tomographic Diffuse Fluorescence Flow Cytometry for Enumeration of Rare Circulating Cells in Vitro and in Vivo.
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Tomographic Diffuse Fluorescence Flow Cytometry for Enumeration of Rare Circulating Cells in Vitro and in Vivo.

机译：体层摄影漫射荧光流式细胞术用于体外和体内枚举稀有循环细胞。

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摘要

Accurate quantification of circulating cell populations is important in many areas of preclinical and clinical biomedical research including the study of metastasized cancers, T-Lymphotocyes and hematopoietic stem cells. Normally this is done either by extraction and analysis of small blood samples or more recently using microscopy-based in vivo fluorescence flow cytometry. In this thesis, a new technological approach to this problem is described using detection of diffuse fluorescent light from relatively large blood vessels in vivo. The 'tomographic diffuse fluorescence flow cytometer' (TDFFC) uses modulated lasers to illuminate a mouse limb and an array of optical fibers coupled to a high-sensitivity photomultiplier tube array operating in photon counting mode to detect weak fluorescence signals from cells. It is first demonstrated that the TDFFC instrument is capable of detecting fluorescent microspheres and Vybrant-DiD labeled cells with excellent accuracy in an optical flow phantom with similar size, optical properties, linear flow rates and autofluorescence as a mouse limb. Preliminary data demonstrating that the TDFFC is capable of detecting circulating cells in nude mice in vivo is also shown. Finally, a number of methods for performing coarse tomographic localization of fluorescent cells within the cross-section of a mouse limb using TDFFC data sets are described, and the feasibility of this approach is demonstrated using in vitro data sets. In principle, this device would allow interrogation of the whole blood volume of a mouse in minutes, with several orders of magnitude sensitivity improvement compared with current approaches.
机译：在临床前和临床生物医学研究的许多领域，包括转移性癌症，T-淋巴细胞和造血干细胞的研究中，循环细胞数量的准确定量非常重要。通常，这可以通过提取和分析少量血液样本来完成，或者最近可以使用基于显微镜的体内荧光流式细胞术来完成。在本文中，描述了一种通过检测来自体内较大血管的漫射荧光来解决该问题的新技术方法。 “层析漫射荧光流式细胞仪”（TDFFC）使用调制激光照射小鼠肢体，并耦合到以光子计数模式运行的高灵敏度光电倍增管阵列的光纤阵列，以检测来自细胞的微弱荧光信号。首次证明了TDFFC仪器能够以与小鼠肢体相似的大小，光学特性，线性流速和自发荧光的光流模型，以优异的精度检测荧光微球和Vybrant-DiD标记的细胞。还显示了证明TDFFC能够在体内检测裸鼠体内循环细胞的初步数据。最后，描述了使用TDFFC数据集对小鼠肢体横截面内的荧光细胞进行粗略层析定位的多种方法，并使用体外数据集证明了该方法的可行性。原则上，该设备将允许在几分钟内询问鼠标的全血量，与当前方法相比，灵敏度提高了几个数量级。

著录项

  • 作者

    Zettergren, Eric William.;

  • 作者单位

    Northeastern University.;

  • 授予单位 Northeastern University.;
  • 学科 Engineering Electronics and Electrical.;Physics Optics.;Engineering Biomedical.
  • 学位 M.S.
  • 年度 2011
  • 页码 72 p.
  • 总页数 72
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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