Advantages of Fresnel biprism-based digital holographic microscopy in quantitative phase imaging
Advantages of Fresnel biprism-based digital holographic microscopy in quantitative phase imaging
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DOI：
10.1117/1.jbo.25.8.086501
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发表时间：
2020-08-01
影响因子：
3.5
通讯作者：
Doblas, Ana
中科院分区：
文献类型：
--
作者：
Hayes-Rounds, Charity;Bogue-Jimenez, Brian;Doblas, Ana
重要性：与其他定量相位成像（QPI）方法相比，数字全息显微术（DHM）的特点是高速，准确性，空间分辨率，时间稳定性和偏振灵敏度（PS）能力。上述特征使DHM适合在广泛的生物学应用中进行实时定量PS相位成像，旨在了解细胞生长和生理过程中发生的动态变化和/或对药剂的反应。目的：菲涅耳双棱镜（FB）的插入在光学显微镜的图像空间中，可能将任何商业系统转变为DHM系统，使QPI具有QPI中的五个期望特征同时：高时间灵敏度、高速度、高精度、高空间分辨率和PS。据我们所知，这是第一个FB为基础的DHM系统提供这五个features all together.Approach：所提出的系统的性能进行了校准与基准相位对象。通过对人U87胶质母细胞瘤细胞的成像，验证了PS的能力。结果：提出的基于FB的DHM系统提供了具有高空间分辨率的精确相位图像。我们的系统的时间稳定性是在几纳米的顺序，使活细胞研究。最后，细胞在不同偏振角下的独特行为（例如，结论：我们已经提出了一种方法，把任何商业光学显微镜与单色照明到PS QPI系统。所提出的系统提供了一个新的，简单的，紧凑的，具有成本效益的格式，由于低成本（几百美元）涉及实现这种简单的架构，使使用这种QPI技术访问大多数实验室与标准的光学显微镜精确的定量PS相位图像。(C)作者。
Significance: The hallmarks of digital holographic microscopy (DHM) compared with other quantitative phase imaging (QPI) methods are high speed, accuracy, spatial resolution, temporal stability, and polarization-sensitivity (PS) capability. The above features make DHM suitable for real-time quantitative PS phase imaging in a broad number of biological applications aimed at understanding cell growth and dynamic changes occurring during physiological processes and/or in response to pharmaceutical agents.Aim: The insertion of a Fresnel biprism (FB) in the image space of a light microscope potentially turns any commercial system into a DHM system enabling QPI with the five desired features in QPI simultaneously: high temporal sensitivity, high speed, high accuracy, high spatial resolution, and PS. To the best of our knowledge, this is the first FB-based DHM system providing these five features all together.Approach: The performance of the proposed system was calibrated with a benchmark phase object. The PS capability has been verified by imaging human U87 glioblastoma cells.Results: The proposed FB-based DHM system provides accurate phase images with high spatial resolution. The temporal stability of our system is in the order of a few nanometers, enabling live-cell studies. Finally, the distinctive behavior of the cells at different polarization angles (e.g., PS capability) can be observed with our system.Conclusions: We have presented a method to turn any commercial light microscope with monochromatic illumination into a PS QPI system. The proposed system provides accurate quantitative PS phase images in a new, simple, compact, and cost-effective format, thanks to the low cost (a few hundred dollars) involved in implementing this simple architecture, enabling the use of this QPI technique accessible to most laboratories with standard light microscopes. (C) The Authors.