Team:ETH Zurich/Process/Validation

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Revision as of 18:58, 21 September 2011

Can you feel the smoke tonight?

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 gradient: Quantification of the gradient in Figure 2: The light intensity of the IPTG-induced GFP signal was quantified by a moving 80&cross;80 moving average. The peak at around 1.2 is due to an air bubble in the channel (see Figure 2).

@@ Line 15: / Line 15: @@
 == System validation for diffusion==
-[[File:Setup_test.png|400px|left|thumb|'''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 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.
-[[File:ETHZ Gradient.png|800px|center|thumb|'''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|'''Quantification of gradient:''' ''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 gradient:''' Quantification of the gradient in Figure 2: The light intensity of the IPTG-induced GFP signal was quantified by a moving 80&cross;80 moving average. The peak at around 1.2 is due to an air bubble in the channel (see Figure 2).]]