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Resolving the depth of fluorescent light by structured illumination and shearing interferometry

机译：通过结构化照明和剪切干涉法解决荧光的深度

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摘要

A method for the depth-sensitive detection of fluorescent light is presented. It relies on a structured illumination restricting the excitation volume and on an interferometric detection of the wave front curvature. The illumination with two intersecting beams of a white-light laser separated in a Sagnac interferometer coupled to the microscope provides a coarse confinement in lateral and axial direction. The depth reconstruction is carried out by evaluating shearing interferograms produced with a Michelson interferometer. This setup can also be used with spatially and temporally incoherent light as emitted by fluorophores. A simulation workflow of the method was developed using a combination of a solution of Maxwell's equations with the Monte Carlo method. These simulations showed the principal feasibility of the method. The method is validated by measurements at reference samples with characterized material properties, locations and sizes of fluorescent regions. It is demonstrated that sufficient signal quality can be obtained for materials with scattering properties comparable to dental enamel while maintaining moderate illumination powers in the milliwatt range. The depth reconstruction is demonstrated for a range of distances and penetration depths of several hundred micrometers.
机译：提出了一种用于荧光的深度敏感检测的方法。它依赖于限制激发体积的结构化照明以及波前曲率的干涉检测。在与显微镜连接的Sagnac干涉仪中分离出的两束白光激光的相交光束，在横向和轴向方向上提供了粗略的限制。通过评估由迈克尔逊干涉仪产生的剪切干涉图来进行深度重建。此设置也可以与荧光团发出的空间和时间不相干光一起使用。该方法的仿真工作流程是通过将麦克斯韦方程组的解决方案与蒙特卡洛方法相结合而开发的。这些仿真表明了该方法的主要可行性。该方法通过对参考样品进行测量而得到验证，这些参考样品具有表征的材料特性，荧光区域的位置和大小。事实证明，对于散射特性可与牙釉质媲美的材料，可以获得足够的信号质量，同时在毫瓦范围内保持适度的照明功率。对于数百微米的距离和穿透深度范围进行了深度重建。

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  • 来源
    《Quantitative phase imaging II》|2016年|97182H.1-97182H.8|共8页
  • 会议地点 San Francisco CA(US)
  • 作者

    Johannes Schindler; Ahmed Elmaklizi; Florian Voit; Ansgar Hohmann; Philipp Schau; Nicole Brodhag; Philipp Krauter; Karsten Frenner; Alwin Kienle; Wolfgang Osten;

  • 作者单位

    Institut fuer Technische Optik, University of Stuttgart, Pfaffenwaldring 9, 70569 Stuttgart, Germany;

    Institut fuer Lasertechnologien in der Medizin und Messtechnik, University of Ulm, Helmholtzstrasse 12, 89801, Ulm, Germany;

    Institut fuer Lasertechnologien in der Medizin und Messtechnik, University of Ulm, Helmholtzstrasse 12, 89801, Ulm, Germany;

    Institut fuer Lasertechnologien in der Medizin und Messtechnik, University of Ulm, Helmholtzstrasse 12, 89801, Ulm, Germany;

    Institut fuer Technische Optik, University of Stuttgart, Pfaffenwaldring 9, 70569 Stuttgart, Germany;

    Institut fuer Technische Optik, University of Stuttgart, Pfaffenwaldring 9, 70569 Stuttgart, Germany;

    Institut fuer Lasertechnologien in der Medizin und Messtechnik, University of Ulm, Helmholtzstrasse 12, 89801, Ulm, Germany;

    Institut fuer Technische Optik, University of Stuttgart, Pfaffenwaldring 9, 70569 Stuttgart, Germany;

    Institut fuer Lasertechnologien in der Medizin und Messtechnik, University of Ulm, Helmholtzstrasse 12, 89801, Ulm, Germany;

    Institut fuer Technische Optik, University of Stuttgart, Pfaffenwaldring 9, 70569 Stuttgart, Germany;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    Fluorescence; Microscopy; Shearing; Interferometry; Structured illumination; Scattering;

    机译：荧光;显微镜；剪切；干涉仪结构化照明；散射;

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