Fabrication of cyclo olefin polymer microfluidic devices for trapping and culturing of yeast cells
| Autor: | Kutlu O. Ulgen, Elif Gencturk, Irem Ezgi Odabasi, Senol Mutlu, Emre Iseri, Sevde Puza |
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| Rok vydání: | 2017 |
| Předmět: |
Materials science
Polymers Microfluidics Saccharomyces cerevisiae Biomedical Engineering Cell Culture Techniques Nanotechnology 02 engineering and technology Cell Separation 01 natural sciences Green fluorescent protein chemistry.chemical_compound Bioreactors Lab-On-A-Chip Devices Doubling time Computer Simulation Molecular Biology Microscopy Chromatography biology Cell growth 010401 analytical chemistry Cycloparaffins Equipment Design Cells Immobilized 021001 nanoscience & nanotechnology biology.organism_classification Fluorescence Yeast 0104 chemical sciences chemistry Hydrodynamics Polystyrene 0210 nano-technology |
| Zdroj: | Biomedical microdevices. 19(2) |
| ISSN: | 1572-8781 |
| Popis: | A microfluidic platform is designed and fabricated to investigate the role of uncharacterized YOR060C (Sld7) protein in aging in yeast cells for the first time. Saccharomyces cerevisiae yeast cells are trapped in the series of C-shaped regions (0.5 nL) of COP (cyclo olefin polymer), PMMA (poly methylmethacrylate), or PS (polystyrene) microbioreactors. The devices are fabricated using hot embossing and thermo-compression bonding methods. Photolithography and electrochemical etching are used to form the steel mold needed for hot embossing. The cell cycle processes are investigated by monitoring green fluorescent protein (GFP) tagged Sld7 expressions under normal as well as calorie restricted conditions. The cells are loaded at 1 μL/min flowrate and trapped successfully within each chamber. The medium is continuously fed at 0.1 μL/min throughout the experiments. Fluorescent signals of the low abundant Sld7 proteins could be distinguished only on COP devices. The background fluorescence of COP is found 1.22 and 7.24 times lower than that of PMMA, and PS, respectively. Hence, experiments are continued with COP, and lasted for more than 40 h without any contamination. The doubling time of the yeast cells are found as 72 min and 150 min, and the growth rates as 9.63 × 10−3 min−1 and 4.62 × 10−3 min−1, in 2% glucose containing YPD and YNB medium, respectively. The product concentration (Sld7p:GFP) increased in accordance with cell growth. The dual role of Sld7 protein in both cell cycle and chronological aging needs to be further investigated following the preliminary experimental results. |
| Databáze: | OpenAIRE |
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