北京大学学报(自然科学版)

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新型多孔单细胞观测芯片设计及在细菌抗药性研究中的应用

姜香丹1,2,欧阳颀1,2,罗春雄1,2   

  1. 1. 北京大学微流与纳米技术研究中心, 北京大学人工微结构与介观物理国家重点实验室, 北京大学物理学院, 北京100871; 2. 北京大学定量生物学中心, 前沿交叉学科研究院, 北京100871;

  • 收稿日期:2013-04-04 出版日期:2014-05-20

Design of Multi-well Based Single Cell Observation Chip and Its Application on Bacteria Antibiotic Property Study

JIANG Xiangdan1,2, OUYANG Qi1,2, LUO Chunxiong1,2   

  1. 1. Center for Microfluidic and Nanotechnology, State Key Laboratory for Artificial Microstructures and Mesoscopic Physics, School of Physics, Peking University, Beijing 100871; 2. Center for Quantitative Biology, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871;

  • Received:2013-04-04 Published:2014-05-20

摘要: 提出一种新型多孔单细胞观测芯片的设计和实现方法, 结合多孔板操作简单和微流控芯片单细胞观测的优势, 为微流控芯片的推广利用提供新思路。在芯片中设计由小孔、特定高度的培养腔室和栅栏组成的并排对称单元, 这三层高度结构把细胞限制在培养室单层固定生长。利用PDMS材料在等离子体处理后对液体的浸润性和对气体吸收的特性, 在小孔加入菌液后细胞很快被自动吸入细胞培养室。只用10 μL液体就可以完成长时间观测细菌所有单层形态变化过程以及高通量并行对比, 实现不同大肠杆菌菌株在不同抗生素浓度下的并行对比分析。此芯片制作的方便性与能在单层细胞程度下实现高通量并行对比的优越性, 使它在大肠杆菌的抗药性以及流式分选后期研究等方面应用前景十分广阔。

关键词: 微流控芯片, 大肠杆菌, 单层生长, 高通量, 抗药性

Abstract: The authors introduced a multi-well based cell observation chip which combined the advantage of easy operation and dynamic observation of single cell. Parallel symmetrical structure unit, which consists of holes, cultivation chambers and barriers, is designed to track cell phenotype changes at single-cell level. Because of the hydrophilism and gas absorption property of the PDMS material after the plasma treatment, the Escherichia coli cells loaded into holes will be soon automatically inhaled into cultivation chambers. Long-term observation under different antibiotic environment and high-throughput parallel comparative study can be realized using only 10 μL liquid. The superiority of such microfluidic chip in realizing monolayer high-throughput parallel study and its convenience may provide many applications, including the antibiotic property study of E. coli and late-stage studies based on fluorescence-activated cell sorting.

Key words: microfluidic, Escherichia coli, single-cell level, high-throughput, antibiotic

中图分类号: 

  • Q143