Multiwavelength Digital Holographic Imaging and Phase Unwrapping of Protozoa Using Custom Fiji Plug-ins
Multiwavelength Digital Holographic Imaging and Phase Unwrapping of Protozoa Using Custom Fiji Plug-ins
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DOI：
10.3389/fphy.2019.00094
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发表时间：
2019-07-05
影响因子：
3.1
通讯作者：
Nadeau, Jay
中科院分区：
文献类型：
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作者：
Cohoe, David;Hanczarek, Iulia;Nadeau, Jay
多波长数字全息显微镜(DHM)已被用于改善数字全息图的位相重建，以减少2pi的位相模糊。然而，大多数用作测试图像的样品都是固体的或附着在表面上，因此很容易确定焦平面和校正色差。在这项研究中，我们将3波长离轴DHM应用于包含不同光谱特征的游泳原生动物，如叶绿素和类胡萝卜素。我们使用角谱方法将全息图再现为幅度和相位图像。从幅度和相位两个方面提出了噪声去除、色差校正和图像配准方法。对位相展开方法进行了评估，并与模拟全息图的预期结果进行了比较。所使用的算法是在使用开源斐济平台的插件中实现的，并且可供使用，大大扩展了可用于DHM的开源软件。
Multiwavelength digital holographic microscopy (DHM) has been used to improve phase reconstructions of digital holograms by reducing 2 pi phase ambiguities. However; most samples used as test images have been solid or adhered to a surface, making it easy to determine focal planes and correct for chromatic aberration. In this study we apply 3-wavelength off-axis DHM to swimming protozoa containing distinct spectral features such as chlorophyll and carotenoids. We reconstruct the holograms into amplitude and phase images using the angular spectrum method. Methods for noise subtraction, chromatic aberration correction, and image registration are presented for both amplitude and phase. Approaches to phase unwrapping are evaluated and compared to expected results from simulated holograms. The algorithms used are implemented in plug-ins using the open source Fiji platform and are available for use, significantly expanding the open-source software available for DHM.