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A microfluidic single-cell array for in situ laminar-flow-based comparative culturing of budding yeast cells
- Source :
- Talanta. 231
- Publication Year :
- 2021
-
Abstract
- 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 × 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 β-estradiol in cell-culturing medium. Experimental results show successful induction of fluorescence in cells immobilized in desired columns that have been dosed with β-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.
- Subjects :
- Yellow fluorescent protein
In situ
biology
Chemistry
Microfluidics
Saccharomyces cerevisiae
Laminar flow
Microfluidic Analytical Techniques
biology.organism_classification
Yeast
Analytical Chemistry
Single-cell analysis
Saccharomycetales
Biophysics
biology.protein
Hydrodynamics
Fluidics
Single-Cell Analysis
Subjects
Details
- ISSN :
- 18733573
- Volume :
- 231
- Database :
- OpenAIRE
- Journal :
- Talanta
- Accession number :
- edsair.doi.dedup.....31f1a7d4455fddca4e1adb2822aab9ac