Team:ETH Zurich/Process/Validation

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Systems Validation
Diffusion-only System A
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System validation for diffusion

Figure 1: Experimental setup for the diffusion test in agarose filled tubes.

To test if we can create a gradient of a small molecule along our agarose filled tube. We filled a tube (2 mm diameter, 7 cm) long with agarose and E. coli with IPTG-inducible GFP was incubated at 37 °C overnight on end of the tube sticking in 1 ml of 10 mM IPTG solution.


Figure 2: GFP gradient in tube: E. coli with IPTG-inducable GFP were incubated in a tube (2 mm diameter, 7 cm long). GFP expression was assessed under the fluorescent microscope after overnight incubation, with a excitation wavelength of 480 nm and a emission wavelength of 510 nm. The 15 microscope photos were reassembled into one using [http://research.microsoft.com/en-us/um/redmond/groups/ivm/ICE/ the Microsoft Research Image Composite Editor].
Figure 3: Quantification of the gradient in Figure 2: The light intensity of the IPTG-induced GFP signal was quantified by a 80×80 pixel moving average. The peak at around 1.2 is due to an air bubble in the channel (see Figure 2).