Poola Praveen Kumar, John Renu
Indian Institute of Technology Hyderabad, Department of Biomedical Engineering, Kandi, Telangana, India.
J Biomed Opt. 2017 Oct;22(10):1-7. doi: 10.1117/1.JBO.22.10.106001.
We report the results of characterization of red blood cell (RBC) structure and its dynamics with nanometric sensitivity using transport of intensity equation microscopy (TIEM). Conventional transport of intensity technique requires three intensity images and hence is not suitable for studying real-time dynamics of live biological samples. However, assuming the sample to be homogeneous, phase retrieval using transport of intensity equation has been demonstrated with single defocused measurement with x-rays. We adopt this technique for quantitative phase light microscopy of homogenous cells like RBCs. The main merits of this technique are its simplicity, cost-effectiveness, and ease of implementation on a conventional microscope. The phase information can be easily merged with regular bright-field and fluorescence images to provide multidimensional (three-dimensional spatial and temporal) information without any extra complexity in the setup. The phase measurement from the TIEM has been characterized using polymeric microbeads and the noise stability of the system has been analyzed. We explore the structure and real-time dynamics of RBCs and the subdomain membrane fluctuations using this technique.
我们报告了使用强度传输方程显微镜（TIEM）以纳米级灵敏度表征红细胞（RBC）结构及其动力学的结果。传统的强度传输技术需要三个强度图像，因此不适用于研究活生物样品的实时动力学。然而，假设样品是均匀的，利用强度传输方程进行相位恢复已通过单次X射线散焦测量得到证明。我们将此技术应用于红细胞等均匀细胞的定量相光显微镜检查。该技术的主要优点是简单、成本效益高，并且易于在传统显微镜上实施。相位信息可以很容易地与常规明场和荧光图像合并，以提供多维（三维空间和时间）信息，而无需在设置上增加任何额外的复杂性。使用聚合物微珠对TIEM的相位测量进行了表征，并分析了系统的噪声稳定性。我们使用该技术探索红细胞的结构和实时动力学以及亚域膜波动。
