Team:Wageningen UR/Project/Devices

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Revision as of 19:35, 19 September 2011

Building a Synchronized Oscillatory System

Custom fluidic device designed by Team Wageningen UR to measure oscillations

Design

The [http://www.nature.com/nature/journal/v463/n7279/abs/nature08753.html paper] oscillatory system is based on, used microfluidic devices to physically constrain the host cells. This was necessary to induce and monitor oscillatory behaviour of a population of e.coli cells. Such microfluidic devices are very expensive and only usable for one experiment. Due to this we set out to find a cheap alternative for these microfluidic devices. Eventually we designed our own flow device, of which the design is seen in figure 1.

Fig.1: Wire frame model of designed flow device.

Bacterial growing platforms

This flow device can accommodate two bacterial growing platforms, a micro-sieve and a micro-dish. The micro-sieve as depicted is figure .. are membranes which have evenly distributed pores of 200 nm in diameter. These membranes are used in dairy industry to sterilise dairy products by removing micro-organism through filtration. Through filtration a cake of cells will form on the membrane, potentially giving us a platform capable of inducing and giving a mean of monitoring oscillatory behaviour of a population of e. coli cells.

Fig.1: Design flow device made in google sketchup

The micro-dish contains porous wells, in which the cells can be physically constrained. It is placed in the socket of the dish, which allows a fluid to be flown over the wells. This fluid can be used to supply the cells with nutrients and wash away excess AHL and cells if necessary. Figure 2 shows a schematic of the device. The inflow and outflow1 can be used to flush the device with fresh medium. The use of outflow2 depends on the bacteria growing platform. In the case of the micro-sieve it can be used to create an under pressure, e.g. with a syringe, to trap the bacteria on the sieve. When using a micro-dish, it can optionally be used to bottom-feed the bacteria in the wells and/or supply additional substances to initalize the oscillatory behaviour of the system, e.g. IPTG in the case of the pRight/lacI hybrid promoter for the streamligned oscillator.

Fig.2: Schematic of the micro-fluidic device depicting the position of in- and outflow

Left: Production of the device in the Wageningen University workshop

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@@ Line 26: / Line 26: @@
 ==== Design ====
-The [http://www.nature.com/nature/journal/v463/n7279/abs/nature08753.html paper] our system is based on used microfluidic devices to measure oscillations. Such microfluidic devices however are very expensive and our iGEM team budget was limited, so we decided to design a customary fluidic device, which was produced in the Wageningen University workshop. To keep as many options as possible open, the design implemented the idea of potentially accommodating two bacteria growing platforms, a micro-sieve and a micro-dish. Gaining more in-depth understanding of our system, we decided to focus on the micro-dish as platform to test the oscillatory behaviour of our construct. Figure 1 and 2 below show the flow chamber as original scetch designed by our team and the 3D rendered depiction of the final device.
+The [http://www.nature.com/nature/journal/v463/n7279/abs/nature08753.html paper] oscillatory system is based on, used microfluidic devices to physically constrain the host cells. This was necessary to induce and monitor oscillatory behaviour of a population of e.coli cells. Such microfluidic devices are very expensive and only usable for one experiment. Due to this we set out to find a cheap alternative for these microfluidic devices. Eventually we designed our own flow device, of which the design is seen in figure 1.
+[[File:mainpic.jpg]]
+'''Fig.1:''' ''Wire frame model of designed flow device.''
+'''Bacterial growing platforms'''
+This flow device can accommodate two bacterial growing platforms, a micro-sieve and a micro-dish. The micro-sieve as depicted is figure .. are membranes which have evenly distributed pores of 200 nm in diameter. These membranes are used in dairy industry to sterilise dairy products by removing micro-organism through filtration. Through filtration a cake of cells will form on the membrane, potentially giving us a platform capable of inducing and giving a mean of monitoring oscillatory behaviour of a population of e. coli cells.
@@ Line 32: / Line 70: @@
 [[File:Scetch device WUR.jpg|400px]] [[File:Module-2 WUR.JPG|333px]]
-'''Fig.1:''' ''Design of the fluidic device: original scetch and 3D render''
+'''Fig.1:''' ''Design flow device made in google sketchup''