Team:ETH Zurich/Process/Validation
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== Results == | == Results == | ||
- | + | Fluorescence pictures of the tube (see Figure 2) showed a clear gradient of the fluorescence signal over approximately 5cm of the tube. After 5cm, the signal strength dropped under the background noise. | |
[[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].]] | ||
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+ | == Analysis == | ||
+ | We quantified the fluorescence signal using a moving average of 80×80 pixel, which moved along the symmetry axis of the tube (see Figure 3). | ||
[[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).]] | ||
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Revision as of 21:05, 21 September 2011
Systems Validation |
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Text goes here |
Experimental SetupTo 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 (see Figure 1). 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. |
ResultsFluorescence pictures of the tube (see Figure 2) showed a clear gradient of the fluorescence signal over approximately 5cm of the tube. After 5cm, the signal strength dropped under the background noise. |
Analysis
We quantified the fluorescence signal using a moving average of 80×80 pixel, which moved along the symmetry axis of the tube (see Figure 3).
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