College of Information Science and Engineering, Fujian Key Laboratory of Light Propagation and Transformation, Huaqiao University, Xiamen, Fujian, China.
Institute of Fluid Physics, China Academy of Engineering Physics, Mianyang, China.
J Biophotonics. 2022 Jul;15(7):e202100400. doi: 10.1002/jbio.202100400. Epub 2022 Mar 21.
Quantitative phase microscopy (QPM) has been emerged as an indispensable diagnostic and characterization tool in biomedical imaging with its characteristic nature of label-free, noninvasive, and real time imaging modality. The integration of holography to the conventional microscopy opens new advancements in QPM featuring high-resolution and quantitative three-dimensional image reconstruction. However, the holography schemes suffer in space-bandwidth and time-bandwidth issues in the off-axis and phase-shifting configuration, respectively. Here, we introduce an on-axis phase-shifting holography based QPM system with single-shot imaging capability. The technique utilizes the Fizeau interferometry scheme in combination with polarization phase-shifting and space-division multiplexing to achieve the single-shot recording of the multiple phase-shifted holograms. Moreover, the high-speed imaging capability with instantaneous recording of spatially phase shifted holograms offers the flexible utilization of the approach in dynamic quantitative phase imaging with robust phase stability. We experimentally demonstrated the validity of the approach by quantitative phase imaging and depth-resolved imaging of paramecium cells. Furthermore, the technique is applied to the phase imaging and quantitative parameter estimation of red blood cells. This integration of a Fizeau-based phase-shifting scheme to the optical microscopy enables a simple and robust tool for the investigations of engineered and biological specimen with real-time quantitative analysis.
定量相位显微镜（QPM）已经成为生物医学成像中不可或缺的诊断和特征工具，具有无标记、非侵入性和实时成像方式的特点。全息术与传统显微镜的结合为 QPM 带来了新的进展，具有高分辨率和定量的三维图像重建功能。然而，全息术方案在离轴和相移配置中分别存在空间带宽和时间带宽的问题。在这里，我们引入了一种具有单次成像能力的基于同轴相移全息术的 QPM 系统。该技术利用菲佐干涉仪方案结合偏振相移和空间分复用，实现了多个相移全息图的单次记录。此外，高速成像能力可瞬时记录空间相移全息图，为动态定量相位成像提供了灵活的应用，具有稳健的相位稳定性。我们通过定量相位成像和草履虫细胞的深度分辨成像实验验证了该方法的有效性。此外，该技术还应用于红细胞的相位成像和定量参数估计。将基于菲佐的相移方案集成到光学显微镜中，为工程和生物样本的实时定量分析提供了简单而强大的工具。
