Yonsei University, School of Mechanical Engineering, Seoul 03722, Republic of Korea.
Yonsei University, School of Integrated Technology & Yonsei Institute of Convergence Technology, Incheon 21983, Republic of Korea.
Methods. 2018 Mar 1;136:66-74. doi: 10.1016/j.ymeth.2017.11.010. Epub 2017 Nov 22.
Color-coded light-emitting diode (LED) microscopy (cLEDscope) is a novel computational microscopy technique capable of multi-contrast and quantitative phase imaging of biological specimens using color-multiplexed illumination. Using specially designed LED patterns, it is capable of recording multiple differential phase contrast (DPC) images in a single exposure and employs a computational algorithm to retrieve the phase distribution of the specimens. Herein, we describe the detailed procedures in the cLEDscope implementation for quantitative phase imaging. Several notable features and caveats in the cLEDscope setup and image processing are also outlined. The imaging model is derived for our specific configuration, and the associated phase-retrieval algorithms are presented on the basis of a weak-object transfer function. As an illustrative application of the quantitative cLEDscope, we demonstrate its utility as a sperm-motility analyzer by exploiting its real-time quantitative imaging capability.
彩色发光二极管（LED）显微镜（cLEDscope）是一种新颖的计算显微镜技术，能够使用彩色复用照明对生物样本进行多对比度和定量相位成像。使用专门设计的 LED 模式，它能够在单次曝光中记录多个微分相差对比度（DPC）图像，并采用计算算法来恢复样本的相位分布。本文详细描述了 cLEDscope 定量相位成像实现的详细步骤。还概述了 cLEDscope 设置和图像处理中的几个显著特点和注意事项。针对我们的特定配置推导出了成像模型，并基于弱物体传递函数提出了相关的相位恢复算法。作为定量 cLEDscope 的说明性应用，我们通过利用其实时定量成像能力，展示了其作为精子活力分析仪的实用性。
