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

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