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Parameter-optimized digital holographic microscope for high-resolution living-cell analysis

机译:参数优化的数字全息显微镜,用于高分辨率活细胞分析

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摘要

A parameter-optimized off-axis setup for digital holographic microscopy is presented for simultaneous, high-resolution, full-field quantitative amplitude and quantitative phase-contrast microscopy and the detection of changes in optical path length in transparent objects, such as undyed living cells. Numerical reconstruction with the described nondiffractive reconstruction method, which suppresses the zero order and the twin image, requires a mathematical model of the phase-difference distribution between the object wave and the reference wave in the hologram plane. Therefore an automated algorithm is explained that determines the parameters of the mathematical model by carrying out the discrete Fresnel transform. Furthermore the relationship between the axial position of the object and the reconstruction distance, which is required for optimization of the lateral resolution of the holographic images, is derived. The lateral and the axial resolutions of the system are discussed and quantified by application to technical objects and to living cells.
机译:提出了一种用于数字全息显微镜的参数优化离轴设置,可用于同时,高分辨率,全场定量振幅和定量相衬显微镜,以及检测透明物体(例如未染色的活细胞)中光程长度的变化。利用所描述的非衍射重建方法的数值重建,其抑制了零级和孪生像,需要在全息图中的物波和参考波之间的相位差分布的数学模型。因此,解释了一种自动算法,该算法通过执行离散菲涅耳变换来确定数学模型的参数。此外,得出了物体的轴向位置和重建距离之间的关系,这是优化全息图像的横向分辨率所需要的。通过应用于技术对象和活细胞,讨论并量化了系统的横向和轴向分辨率。

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