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Induction of sustained glycolytic oscillations in single yeast cells using microfluidics and optical tweezers

Paper i proceeding
Författare Anna-Karin Gustavsson
Caroline B. Adiels
Mattias Goksör
Publicerad i Proc. SPIE, 8458 s. 84580Y
Volym 8458
Sidor 84580Y
Publiceringsår 2012
Publicerad vid Institutionen för fysik (GU)
Sidor 84580Y
Språk en
Länkar dx.doi.org/10.1117/12.928919
Ämnesord Cell heterogeneity, glycolysis, limit-cycle oscillations, microfluidic flow chamber, optical manipulation
Ämneskategorier Optik, Optisk fysik, Cellbiologi

Sammanfattning

Yeast glycolytic oscillations have been studied since the 1950s in cell free extracts and in intact cells. Until recently, sustained oscillations have only been observed in intact cells at the population level. The aim of this study was to investigate sustained glycolytic oscillations in single cells. Optical tweezers were used to position yeast cells in arrays with variable cell density in the junction of a microfluidic flow chamber. The microfluidic flow chambers were fabricated using soft lithography and the flow rates in the different inlet channels were individually controlled by syringe pumps. Due to the low Reynolds number, the solutions mixed by diffusion only. The environment in the junction of the chamber could thus be controlled by changing the flow rates in the inlet channels, with a complete change of environment within 2 s. The optimum position of the cell array was determined by simulations, to ensure complete coverage of the intended solution without any concentration gradients over the cell array. Using a DAPI filter set, the NADH auto fluorescence could be monitored in up to 100 cells simultaneously. Sustained oscillations were successfully induced in individual, isolated cells within specific flow rates and concentrations of glucose and cyanide. By changing the flow rates without changing the surrounding solution, it was found that the cell behavior was dependent on the concentration of chemicals in the medium rather than the flow rates in the range tested. Furthermore, by packing cells tightly, cell-to-cell interaction and synchronization could be studied.

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