Abstract Reactive oxygen species (ROS) are formed by several different reactions and play a major role in cell cycling and as a messenger in normal cell transduction and have diverse biological effects when high in concentration. ROS may trigger DNA damage, cell mutation, inflammation, cell proliferation, and ultimately they may induce carcinogenesis. This study aims to design, fabricate, and develop a novel on-chip ROS detection based on mounted in-house built fluorescence microscope in microfluidics, which has not been reported previously. An in-house 3D printed built digital portable fluorescence type microscope for on-chip ROS detection has been developed. The combined use of the fluorescence microscope and a glass-based microfluidic chip both enabled to provide high-quality images of the living cells with ease of controlling and handling of the biological sample. As an evidence of the fluorescence microscope, breast cancer cells were loaded with 2′,7′-dichlorofluorescein diacetate (DCFDA, 50 μM) for 45 min on-chip which produces green fluorescence upon reaction with intracellular ROS. Fluorescence of the MCF-7 cells has been captured to obtain a profile of ROS generation for 1 day. LabVIEW 2019 was used along with its high-performance image analysis and IMAQ Vision tools. The viability of the cells was performed by LIVE/DEAD assay and Luna cell counter. The developed platform provides real-time monitoring and quantification of ROS. A strong linear correlation between concentrations versus fluorescence has been recorded. This result recommends that the current microfluidic chip provides an in-vitro platform for the assessment of cell cycle progression and evaluation of the variety of drugs.

中文翻译：

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通过内部构建的荧光显微镜在芯片上实时检测和量化 MCF-7 细胞中的活性氧

摘要 活性氧（ROS）由多种不同的反应形成，在细胞循环中起主要作用，在正常细胞转导中作为信使，在高浓度时具有多种生物学效应。ROS 可能引发 DNA 损伤、细胞突变、炎症、细胞增殖，最终可能诱发癌变。本研究旨在设计、制造和开发一种基于微流体中安装的内部内置荧光显微镜的新型片上 ROS 检测，这在以前没有报道过。已经开发了用于片上 ROS 检测的内部 3D 打印内置数字便携式荧光型显微镜。荧光显微镜和基于玻璃的微流控芯片的结合使用都能够提供高质量的活细胞图像，并且易于控制和处理生物样品。作为荧光显微镜的证据，乳腺癌细胞在芯片上加载 2',7'-二氯荧光素二乙酸酯（DCFDA，50 μM）45 分钟，在与细胞内 ROS 反应时产生绿色荧光。已捕获 MCF-7 细胞的荧光以获得 1 天的 ROS 生成概况。LabVIEW 2019 与其高性能图像分析和 IMAQ Vision 工具一起使用。通过 LIVE/DEAD 测定和 Luna 细胞计数器进行细胞活力。开发的平台提供了 ROS 的实时监控和量化。已记录浓度与荧光之间的强线性相关性。这一结果表明，目前的微流控芯片为细胞周期进程的评估和多种药物的评价提供了一个体外平台。