Tsai Chen-Ming, Vyas Sunil, Luo Yuan
Opt Express. 2024 Feb 26;32(5):7919-7930. doi: 10.1364/OE.514225.
Digital holographic microscopy (DHM) is a powerful quantitative phase imaging (QPI) technique that is capable of recording sample's phase information to enhance image contrast. In off-axis DHM, high-quality QPI images can be generated within a single recorded hologram, and the system stability can be enhanced by common-path configuration. Diffraction gratings are widely used components in common-path DHM systems; however, the presence of multiple diffraction beams leads to system power loss. Here, we propose and demonstrate implementation of a volume holographic grating (VHG) in common-path DHM, which provides single diffraction order. VHG in common-path DHM (i.e., VHG-DHM) helps in improving signal-to-noise ratio as compared to the conventional DHM. In addition, VHG, with inherently high angular selectivity, reduces image noise caused by stray light. With a simple fabrication process, it is convenient to utilize VHG to control the beam separation angle of DHM. Further, by using Bragg-matched wavelength degeneracy to avoid potential cell damaging effect in blue light, the VHG is designed for recording at a maximum sensitive wavelength of ∼488 nm, while our VHG-DHM is operated at the longer wavelength of red 632.8 nm for cell observation. Experimental results, measured by the VHG-DHM, show the measurement of target thickness ranging from 100 nm to 350 nm. In addition, stability of the system is quantitatively measured. High-contrast QPI images of human lung cancer cells are demonstrated.
数字全息显微镜(DHM)是一种强大的定量相位成像(QPI)技术,能够记录样品的相位信息以增强图像对比度。在离轴DHM中,高质量的QPI图像可以在单个记录的全息图内生成,并且通过共光路配置可以提高系统稳定性。衍射光栅是共光路DHM系统中广泛使用的组件;然而,多个衍射光束的存在会导致系统功率损失。在此,我们提出并展示了在共光路DHM中实现体全息光栅(VHG),它提供单一衍射级次。共光路DHM中的VHG(即VHG-DHM)与传统DHM相比有助于提高信噪比。此外,具有固有高角度选择性的VHG可降低由杂散光引起的图像噪声。通过简单的制造工艺,利用VHG来控制DHM的光束分离角很方便。此外,通过使用布拉格匹配波长简并来避免蓝光中潜在的细胞损伤效应,VHG设计用于在约488 nm的最大敏感波长处记录,而我们的VHG-DHM在红色632.8 nm的较长波长下运行用于细胞观察。由VHG-DHM测量的实验结果显示了目标厚度在100 nm至350 nm范围内的测量。此外,对系统的稳定性进行了定量测量。展示了人肺癌细胞的高对比度QPI图像。