Zhang Xu, Sun Rui, Gong Pengfei, Al-Olayan Ebtesam, Abukhadra Mostafa R, Liu Bo, Su Ping, Zhang Dawei, Feng Shilun
Engineering Research Center of Optical Instrument and System, the Ministry of Education, Shanghai Key Labora-tory of Modern Optical System, University of Shanghai for Science and Technology, Shanghai, 200093, China; Xiangfu Laboratory, Jiaxing Key Laboratory of Bio-semiconductor, Jiashan, 314102, China; State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai, 200050, China.
Xiangfu Laboratory, Jiaxing Key Laboratory of Bio-semiconductor, Jiashan, 314102, China; State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai, 200050, China; Nano-translational Medicine Research Center, Yangtze Delta Region Institute of Tsinghua University, Zhejiang, Jiaxing, 314000, China.
Talanta. 2025 Mar 1;284:127242. doi: 10.1016/j.talanta.2024.127242. Epub 2024 Nov 19.
Droplet microfluidic chips have emerged as an efficient platform for single-cell analysis due to their high sensitivity, efficiency, and throughput, showing significant potential in pathogen detection. However, current droplet microfluidic chips encounter challenges in large-scale droplet quantification and precise imaging, rendering them unsuitable for the high-throughput pathogen detection required for a large number of samples. To address these issues, this study developed a high-precision fluorescence imaging system utilizing a confocal reflective fluorescence approach, which is an advanced microscopy technique that combines confocal microscopy and reflected fluorescence imaging. It can obtain fluorescence signal and reflected light signal in the sample at the same time, so as to provide richer and more comprehensive image information. The system offers a large field of view (17.8 mm × 17.8 mm) and high resolution (20 μm), enabling the rapid imaging of 30,000 droplets within 10 s, thereby significantly enhancing detection efficiency and automation. Additionally, the enzymatic reaction of Escherichia coli (E. coli) was implemented using the droplet microfluidic chip to validate the effectiveness of the optical imaging system, with results demonstrating the system's capability to accurately capture fluorescence changes during the reaction.
液滴微流控芯片因其高灵敏度、高效率和高通量，已成为单细胞分析的有效平台，在病原体检测方面显示出巨大潜力。然而，目前的液滴微流控芯片在大规模液滴定量和精确成像方面面临挑战，使其不适用于大量样品所需的高通量病原体检测。为了解决这些问题，本研究开发了一种利用共焦反射荧光方法的高精度荧光成像系统，这是一种将共聚焦显微镜和反射荧光成像相结合的先进显微镜技术。它可以同时获取样品中的荧光信号和反射光信号，从而提供更丰富、更全面的图像信息。该系统具有大视野（17.8毫米×17.8毫米）和高分辨率（20微米），能够在10秒内对30000个液滴进行快速成像，从而显著提高检测效率和自动化程度。此外，利用液滴微流控芯片进行了大肠杆菌的酶促反应，以验证光学成像系统的有效性，结果表明该系统能够准确捕捉反应过程中的荧光变化。
