Institute of Biomedical Electronics and Bioinformatics, National Taiwan University, Taipei, Taiwan, ROC.
J Biophotonics. 2013 May;6(5):416-24. doi: 10.1002/jbio.201200022. Epub 2012 Aug 24.
Quantification of three-dimensional (3D) refractive index (RI) with sub-cellular resolution is achieved by digital holographic microtomography (DHμT) using quantitative phase images measured at multiple illumination angles. The DHμT system achieves sensitive and fast phase measurements based on iterative phase extraction algorithm and asynchronous phase shifting interferometry without any phase monitoring or active control mechanism. A reconstruction algorithm, optical diffraction tomography with projection on convex sets and total variation minimization, is implemented to substantially reduce the number of angular scattered fields needed for reconstruction without sacrificing the accuracy and quality of the reconstructed 3D RI distribution. Tomogram of a living CA9-22 cell is presented to demonstrate the performance of the method. Further, a statistical analysis of the average RI of the nucleoli, the nucleus excluding the nucleoli and the cytoplasm of twenty CA9-22 cells is performed.
利用数字全息显微层析术(DHμT),通过在多个照明角度下测量定量相位图像,实现了具有亚细胞分辨率的三维(3D)折射率(RI)的量化。DHμT 系统基于迭代相位提取算法和异步相移干涉术实现了灵敏快速的相位测量,无需任何相位监测或主动控制机制。实施了一种重建算法,即基于凸集上的光衍射层析术和全变差最小化,可大大减少重建所需的角散射场数量,而不会牺牲重建的 3D RI 分布的准确性和质量。展示了一个活 CA9-22 细胞的断层扫描图像,以证明该方法的性能。此外,对二十个 CA9-22 细胞的核仁、核仁以外的核和细胞质的平均 RI 进行了统计分析。