Abstract
Spatial gradients of diffusible signalling molecules play crucial roles in controlling diverse cellular behaviour such as cell differentiation, tissue patterning and chemotaxis. In this paper, we report the design and testing of a microfluidic device for diffusionbased gradient generation for cellular assays. A unique channel design of the device eliminates cross-flow between the source and sink channels, thereby stabilising gradients by passive diffusion. The platform also enables quick and flexible control of chemical concentration that makes highly dynamic gradients in diffusion chambers. A model with the first approximation of diffusion and surface adsorption of molecules recapitulates the experimentally observed gradients. Budding yeast cells cultured in a gradient of a chemical inducer expressed a reporter fluorescence protein in a concentration dependent manner. This microfluidic platform serves as a versatile prototype applicable to a broad range of biomedical investigations.
| Original language | English |
|---|---|
| Pages (from-to) | 132-145 |
| Number of pages | 14 |
| Journal | Physica. A, Statistical Mechanics and its Applications |
| Volume | 470 |
| Early online date | 5 Dec 2016 |
| DOIs | |
| Publication status | Published - 15 Mar 2017 |
Bibliographical note
AcknowledgementsWe thank Marco Thiel and Alessandro De Moura for helpful discussions and advice, Stefan Hoppler and Mamen Romano for critical reading of the manuscript, James Hislop for his help with the plasma cleaner, Alex Brand for the microscopy imaging system and Alistair Robertson for fabricating the hydrostatic flow controllers. We also thank Diane Massie and Yvonne Turnbull for technical assistance. This work was supported by Scottish Universities Life Sciences Alliance (SULSA)and the University of Aberdeen .
Keywords
- Microfluidics
- Gradient
- Diffusion
- Yeast
- Saccharomyces cerevisiae
