The Maharaja Sayajirao University of Baroda, Faculty of Technology and Engineering, Department of Ap, India.
Uka Tarsadia University, Department of Physics, Bardoli, Gujarat, India.
J Biomed Opt. 2017 Dec;22(12):1-11. doi: 10.1117/1.JBO.22.12.126001.
Quantitative three-dimensional (3-D) imaging of living cells provides important information about the cell morphology and its time variation. Off-axis, digital holographic interference microscopy is an ideal tool for 3-D imaging, parameter extraction, and classification of living cells. Two-beam digital holographic microscopes, which are usually employed, provide high-quality 3-D images of micro-objects, albeit with lower temporal stability. Common-path digital holographic geometries, in which the reference beam is derived from the object beam, provide higher temporal stability along with high-quality 3-D images. Self-referencing geometry is the simplest of the common-path techniques, in which a portion of the object beam itself acts as the reference, leading to compact setups using fewer optical elements. However, it has reduced field of view, and the reference may contain object information. Here, we describe the development of a common-path digital holographic microscope, employing a shearing plate and converting one of the beams into a separate reference by employing a pin-hole. The setup is as compact as self-referencing geometry, while providing field of view as wide as that of a two-beam microscope. The microscope is tested by imaging and quantifying the morphology and dynamics of human erythrocytes.
定量三维（3-D）成像活体细胞提供了关于细胞形态及其时间变化的重要信息。离轴数字全息干涉显微镜是 3-D 成像、参数提取和活细胞分类的理想工具。虽然双光束数字全息显微镜提供了微物体的高质量 3-D 图像，但时间稳定性较低。共光路数字全息几何形状，其中参考光束来自物体光束，提供了更高的时间稳定性以及高质量的 3-D 图像。自参考几何形状是最简单的共光路技术之一，其中物体光束的一部分自身充当参考，从而使用较少的光学元件实现紧凑的设置。然而，它的视场减小，并且参考可能包含物体信息。在这里，我们描述了一种共光路数字全息显微镜的开发，该显微镜采用剪切板，并通过使用小孔将其中一个光束转换为单独的参考。该设置与自参考几何形状一样紧凑，同时提供与双光束显微镜一样宽的视场。该显微镜通过对人红细胞的形态和动力学进行成像和量化来进行测试。
