Journal
TALANTA
Volume 231, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.talanta.2021.122401
Keywords
Microfluidics; Cell trapping; Single-cell analysis; Comparative cell culturing; Saccharomyces cerevisiae
Categories
Funding
- National Natural Science Foundation of China [61774036, 31771299]
- National Key R&D Program of China [2018YFF01012100]
- Science and Technology Project of Jiangsu Province [BZ2020067]
- Fundamental Research Funds for the Central Universities
- Open Research Fund of State Key Laboratory of Bioelectronics
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The MiSCA microfluidic single-cell array allows for immobilizing yeast cells in a precisely controlled microenvironment and conducting comparative culturing under the influence of different media and compounds. This system enables parallel sets of experiments and control experiments, as well as comparative experiments under well-defined laminar-perfusion conditions.
To facilitate in situ comparative culturing of budding yeast cells in a precisely controlled microenvironment, we developed a microfluidic single-cell array (MiSCA) with 96 traps (16 rows x 6 columns) for single-cell immobilization. Through optimization of the distances between neighboring traps and the applied flow rates by using a hydraulic equivalent circuit of the fluidic network, yeast cells were delivered to each column of the array by laminar focused flows and reliably captured at the traps by hydrodynamic forces with about 90% efficiency of cell immobilization. Immobilized cells in different columns within the same device can then be cultured in parallel while being exposed to different media and compounds delivered by laminar flows. For biological validation of the comparative cell-culturing device, we used budding yeast that can express yellow fluorescent protein upon the addition of beta-estradiol in cell-culturing medium. Experimental results show successful induction of fluorescence in cells immobilized in desired columns that have been dosed with beta-estradiol. The MiSCA system allows for performing sets of experiments and control experiments in parallel in the same device, or for executing comparative experiments under well-defined laminar-perfusion conditions with different media, as well as in situ monitoring of dynamic cellular responses upon different analytical compounds or reagents for single-cell analysis.
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