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首页> 外文期刊>Journal of biomedical optics >Electron multiplying charge-coupled device-based fluorescence cross-correlation spectroscopy for blood velocimetry on zebrafish embryos
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Electron multiplying charge-coupled device-based fluorescence cross-correlation spectroscopy for blood velocimetry on zebrafish embryos

机译：基于电子倍增电荷耦合器件的荧光互相关光谱用于斑马鱼胚胎的血液测速

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摘要

Biomedical issues in vasculogenesis and cardiogenesis require methods to follow hemodynamics with high spatial (micrometers) and time (milliseconds) resolution. At the same time, we need to follow relevant morphogenetic processes on large fields of view. Fluorescence cross-correlation spectroscopy coupled to scanning or wide-field microscopy meets these needs but has limited flexibility in the excitation pattern. To overcome this limitation, we develop here a two-photon two-spots setup coupled to an all-reflective near-infrared (NIR) optimized scanning system and to an electron multiplying charge-coupled device. Two NIR laser spots are spaced at adjustable micron-size distances (1 to 50 μm) by means of a Twyman-Green interferometer and repeatedly scanned on the sample, allowing acquisition of information on flows at 4 ms-3 μm time-space resolution in parallel on an extended field of view. We analyze the effect of nonhomogeneous and variable flow on the cross-correlation function by numerical simulations and show exemplary application of this setup in studies of blood flow in zebrafish embryos in vivo. By coupling the interferometer with the scanning mirrors and by computing the cross-correlation function of fluorescent red blood cells, we are able to map speed patterns in embryos' vessels.
机译：血管生成和心脏生成中的生物医学问题需要采用具有高空间（微米）和时间（毫秒）分辨率的血液动力学追踪方法。同时，我们需要在大视野中遵循相关的形态发生过程。荧光互相关光谱仪与扫描或宽视场显微镜相结合可以满足这些需求，但激发模式的灵活性有限。为了克服这一限制，我们在这里开发了一种双光子两点设置，该设置与全反射近红外（NIR）优化的扫描系统以及电子倍增电荷耦合器件耦合。通过Twyman-Green干涉仪将两个NIR激光光斑以可调的微米尺寸距离（1到50μm）隔开，并在样品上重复扫描，从而可以以4 ms-3μm的时空分辨率获取有关流的信息。平行于扩展视野。我们通过数值模拟分析了非均匀流动和可变流动对互相关函数的影响，并显示了该设置在研究斑马鱼胚胎体内血流方面的示例性应用。通过将干涉仪与扫描镜耦合，并通过计算荧光红细胞的互相关函数，我们能够绘制出胚胎血管中的速度模式。

著录项

  • 来源
    《Journal of biomedical optics》 |2014年第6期|067007.1-067007.12|共12页
  • 作者

    Paolo Pozzi; Laura Sironi; Laura DAlfonso; Margaux Bouzin; Maddalena Collini; Giuseppe Chirico; Piersandro Pallavicini; Franco Cotelli; Efrem A. Foglia;

  • 作者单位

    Universita degli Studi di Milano-Bicocca, Dipartimento di Fisica, Piazza della Scienza 3, I-20126, Milano, Italy;

    Universita degli Studi di Milano-Bicocca, Dipartimento di Fisica, Piazza della Scienza 3, I-20126, Milano, Italy;

    Universita degli Studi di Milano-Bicocca, Dipartimento di Fisica, Piazza della Scienza 3, I-20126, Milano, Italy;

    Universita degli Studi di Milano-Bicocca, Dipartimento di Fisica, Piazza della Scienza 3, I-20126, Milano, Italy;

    Universita degli Studi di Milano-Bicocca, Dipartimento di Fisica, Piazza della Scienza 3, I-20126, Milano, Italy;

    Universita degli Studi di Milano-Bicocca, Dipartimento di Fisica, Piazza della Scienza 3, I-20126, Milano, Italy;

    Universita di Pavia, Dipartimento di Chimica, viale Taramelli, 12 - 27100 Pavia, Italy;

    Universita degli Studi di Milano, Dipartimento di Bioscienze, Via Celoria 26, I-20133, Milano, Italy;

    Universita degli Studi di Milano, Dipartimento di Bioscienze, Via Celoria 26, I-20133, Milano, Italy;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    fluorescence correlation; velocimetry; zebrafish; hemodynamics; microfluidics;

    机译：荧光相关测速斑马鱼血液动力学微流体;

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