Team:ETH Zurich/Process/Validation

[[File:Setup_test.png|400px|left|thumb|'''Figure 1: Experimental setup for the diffusion test in agarose filled tubes.''' ]]

[[File:Setup_test.png|400px|left|thumb|'''Figure 1: Experimental setup for the diffusion test in agarose filled tubes.''' ]]

To validate that 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''. The E. coli cells were engineered to express an IPTG-inducible GFP. The cells were incubated at 37 °C overnight. One end of the tube was connected to a sample medium (1 ml) containing 10 mM IPTG solution.  

To validate that 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''. The E. coli cells were engineered to express an IPTG-inducible GFP. The cells were incubated at 37 °C overnight. One end of the tube was connected to a sample medium (1 ml) containing 10 mM IPTG solution.  

[[File:ETHZ Gradient.png|800px|center|thumb|'''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].]]

[[File:ETHZ Gradient.png|800px|center|thumb|'''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].]]

[[File:Quantification.png|800px|center|thumb|'''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 1.2cm is due to an air bubble in the channel (see Figure 2).]]

[[File:Quantification.png|800px|center|thumb|'''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 1.2cm is due to an air bubble in the channel (see Figure 2).]]