Team:ETH Zurich/Process/Validation

From 2011.igem.org

(Difference between revisions)
Line 6: Line 6:
|-
|-
|style="border-left: none;"|[[#Experimental Setup|Experimental Setup]]
|style="border-left: none;"|[[#Experimental Setup|Experimental Setup]]
-
|[[#Results |Results ]] [[#Analysis |Analysis]]
+
|[[#Results |Results ]]
 +
|[[#Analysis |Analysis]]
|}  
|}  
|-
|-
-
|colspan="2"|'''This page presents a biological experiment which showed that the setup of the flow channel works as predicted by the models.'''
+
|colspan="2"|'''This page presents a biological experiment which showed that the setup of the flow channel works as predicted by the models (compare plots in the [[Team:ETH_Zurich/Modeling/Microfluidics#Simulation|modeling section]]).'''
|}
|}
{|class="roundContainer"
{|class="roundContainer"

Revision as of 21:23, 21 September 2011

Menu image preload Menu image preload Menu image preload Menu image preload Menu image preload Menu image preload


Systems Validation
Experimental Setup Results Analysis
This page presents a biological experiment which showed that the setup of the flow channel works as predicted by the models (compare plots in the modeling section).

Experimental Setup

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 (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.

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.

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].

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).

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).
Retrieved from "http://2011.igem.org/Team:ETH_Zurich/Process/Validation"