Accurate quantitative phase digital holographic microscopy with single- and multiple-wavelength telecentric and nontelecentric configurations.
Accurate quantitative phase digital holographic microscopy with single- and multiple-wavelength telecentric and nontelecentric configurations.
复制标题
DOI：
10.1364/ao.55.005666
复制
发表时间：
2016-07
期刊：
影响因子：
1.9
通讯作者：
Thanh Nguyen;G. Nehmetallah;C. Raub;S. Mathews;R. Aylo
中科院分区：
文献类型：
--
作者：
Thanh Nguyen;G. Nehmetallah;C. Raub;S. Mathews;R. Aylo
在这项工作中，我们从理论上和实验上研究了在透射和反射模式下使用远心和非远心配置的单波长和多波长数字全息显微镜（DHM）。一种单波长远心成像系统在DHM最初提出，以避免由于显微镜物镜（MO）在标准的非远心DHM配置的残余抛物线相位畸变。然而，远心结构不能补偿高阶相位像差。作为单波长远心DHM （SW-DHM）的远心和非远心安排的延伸，我们提出了反射和传输模式的多波长远心DHM （MW-TDHM）。MW-TDHM结构的优点是在无相位模糊的情况下扩展了垂直测量范围，光学上消除了传统MW-DHM中由MO引起的抛物线相位畸变。这些配置消除了需要第二个参考全息图来减去两相图，使数字自动像差补偿比非远心配置更容易应用。我们还讨论了一种利用空间滤波消除零阶图像和虚图像的重建算法，以及另一种利用apodiization最小化波动强度的算法。此外，我们采用两个多项式模型，使用二维表面拟合来数字补偿色差（在多波长情况下）和高阶相位像差。采用自定义开发的用户友好图形用户界面来自动化所有配置的重建过程。最后，TDHM用于可视化高侵袭性MDA-MB-231培养的乳腺癌细胞的细胞。
In this work, we investigate, both theoretically and experimentally, single-wavelength and multiwavelength digital holographic microscopy (DHM) using telecentric and nontelecentric configurations in transmission and reflection modes. A single-wavelength telecentric imaging system in DHM was originally proposed to circumvent the residual parabolic phase distortion due to the microscope objective (MO) in standard nontelecentric DHM configurations. However, telecentric configurations cannot compensate for higher order phase aberrations. As an extension to the telecentric and nontelecentric arrangements in single-wavelength DHM (SW-DHM), we propose multiple-wavelength telecentric DHM (MW-TDHM) in reflection and transmission modes. The advantages of MW-TDHM configurations are to extend the vertical measurement range without phase ambiguity and optically remove the parabolic phase distortion caused by the MO in traditional MW-DHM. These configurations eliminate the need for a second reference hologram to subtract the two-phase maps and make digital automatic aberration compensation easier to apply compared to nontelecentric configurations. We also discuss a reconstruction algorithm that eliminates the zero-order and virtual images using spatial filtering and another algorithm that minimizes the intensity of fluctuations using apodization. In addition, we employ two polynomial models using 2D surface fitting to compensate digitally for chromatic aberration (in the multiwavelength case) and for higher order phase aberrations. A custom-developed user-friendly graphical user interface is employed to automate the reconstruction processes for all configurations. Finally, TDHM is used to visualize cells from the highly invasive MDA-MB-231 cultured breast cancer cells.