A Large-field droplets for high-throughput Escherichia coli identification within one field of view

Xu Zhang; Rui Sun; Pengfei Gong; Ebtesam Al-Olayan; Mostafa R Abukhadra; Bo Liu; Ping Su; Dawei Zhang; Shilun Feng

doi:10.1016/j.talanta.2024.127242

A Large-field droplets for high-throughput Escherichia coli identification within one field of view

Talanta. 2024 Nov 19:284:127242. doi: 10.1016/j.talanta.2024.127242. Online ahead of print.

Authors

Xu Zhang¹, Rui Sun², Pengfei Gong³, Ebtesam Al-Olayan⁴, Mostafa R Abukhadra⁵, Bo Liu⁶, Ping Su⁷, Dawei Zhang⁸, Shilun Feng⁹

Affiliations

¹ 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.
² 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.
³ Xiangfu Laboratory, Jiaxing Key Laboratory of Bio-semiconductor, Jiashan, 314102, China.
⁴ Department of Zoology, College of Science, King Saud University, Riyadh, Saudi Arabia.
⁵ Materials Technologies and their applications Lab, Faculty of Science, Beni-Suef University, Beni Suef city, Egypt.
⁶ Xiangfu Laboratory, Jiaxing Key Laboratory of Bio-semiconductor, Jiashan, 314102, China; School of Microelectronics, Shanghai University, Shanghai, 200444, China.
⁷ Shenzhen International Graduate School, Tsinghua University, Shenzhen, 518055, China.
⁸ 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. Electronic address: dwzhang@usst.edu.cn.
⁹ State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai, 200050, China. Electronic address: shilun.feng@mail.sim.ac.cn.

PMID: 39579491
DOI: 10.1016/j.talanta.2024.127242

Abstract

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.

Keywords: E. coli; Enzymatic reaction; Fluorescence imaging; Large field of view; Microfluidic chip.