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Fabrication of cyclo olefin polymer microfluidic devices for trapping and culturing of yeast cells.
Puza, Sevde; Gencturk, Elif; Odabasi, Irem E; Iseri, Emre; Mutlu, Senol; Ulgen, Kutlu O.
Afiliación
  • Puza S; Department of Chemical Engineering, Biosystems Engineering Laboratory, Bogazici University, 34342, Istanbul, Turkey.
  • Gencturk E; Department of Chemical Engineering, Biosystems Engineering Laboratory, Bogazici University, 34342, Istanbul, Turkey.
  • Odabasi IE; Department of Chemical Engineering, Biosystems Engineering Laboratory, Bogazici University, 34342, Istanbul, Turkey.
  • Iseri E; Department of Electrical and Electronics Engineering, BUMEMS Laboratory, Bogazici University, 34342, Istanbul, Turkey.
  • Mutlu S; Department of Electrical and Electronics Engineering, BUMEMS Laboratory, Bogazici University, 34342, Istanbul, Turkey.
  • Ulgen KO; Department of Chemical Engineering, Biosystems Engineering Laboratory, Bogazici University, 34342, Istanbul, Turkey. ulgenk@boun.edu.tr.
Biomed Microdevices ; 19(2): 40, 2017 Jun.
Article en En | MEDLINE | ID: mdl-28466286
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Polímeros / Saccharomyces cerevisiae / Separación Celular / Células Inmovilizadas / Técnicas de Cultivo de Célula / Cicloparafinas / Dispositivos Laboratorio en un Chip Idioma: En Revista: Biomed Microdevices Asunto de la revista: ENGENHARIA BIOMEDICA Año: 2017 Tipo del documento: Article País de afiliación: Turquía Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Polímeros / Saccharomyces cerevisiae / Separación Celular / Células Inmovilizadas / Técnicas de Cultivo de Célula / Cicloparafinas / Dispositivos Laboratorio en un Chip Idioma: En Revista: Biomed Microdevices Asunto de la revista: ENGENHARIA BIOMEDICA Año: 2017 Tipo del documento: Article País de afiliación: Turquía Pais de publicación: Estados Unidos