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Team:ETH Zurich/expresults/diffusion
From 2014.igem.org
Diffusion On Chip
Our project aims for the biological implementation of cellular automata with XOR logic gates. In order to achieve this, we found a way to create a regular grid of cells with a defined, optimal neighborhood. This means channel length, well size, and medium were optimized and the properties were modelled with Matlab and Comsol whenever feasible. With these in silico results in mind we used CAD software to design our custom made molds, which where then 3D-printed and used for the production of PDMS chips. The cells containing one of our genetic circuits were encapsulated in alginate beads and loaded on the millifluidic chip. This approach allowed us to establish a method for measuring diffusion and cell-to-cell communication. In particular, a step towards the emergence of complex patters by cell-to-cell communication was made. Also the Comsol model regarding pattern formation was confirmed experimentally with our rapid-prototyping approach. The final time-lapse video of the cell-to-cell communication experiment is shown below in video 1.
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| Video 1 Row wise, self-propagating cell-to-cell communication of E. coli cells confined in alginate beads (d=3 mm) on a custom-made millifluidic PDMS chip. All cells contained [http://parts.igem.org/Part:BBa_J23100 constitutively (BBa_J23100)] expressed [http://parts.igem.org/Part:BBa_C0062 LuxR (BBa_C0062)], and riboregulated sfGFP followed by [http://parts.igem.org/Part:BBa_C0161 LuxI (BBa_C0161)] , together under the control of the [http://parts.igem.org/Part:BBa_R0062 pLux promoter (BBa_R0062)]. LuxI catalyzes the production of the autoinducer 3OC6-HSL, which is then diffusing from cell to cell. For initialization, the cells in one bead of the top row were induced with 3OC6-HSL before encapsulation. Imaging was implemented with a Biostep Dark-Hood DH-50 (Argus X1 software) fitted with a Canon EOS 500D DSLR camera and a fluorescence filter (545 nm filter). Pictures were usually taken every 2 min at an excitation wavelength (470 nm) with the standard Canon EOS Utility software. Time-lapse movies were created with Adobe After Effects CC software. |
