Team:Grenoble-EMSE-LSU/Project/Validation

From 2013.igem.org

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<h1>Validation</h1>
<h1>Validation</h1>
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<p> We ran an experiment proving the stabilization of a bacterial culture, the kinetic lasts for 12 hours, and the concentration of bacterial was stabilized for 5 consecutive hours.
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<p> To validate our heuristic method, we ran two experiments where we tried to stabilize the concentration of living cells in a bacterial culture during 5 consecutive hours. Both were successful, and we will report in more detail one of them. </p>
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<h1>Experiment</h1>
<h1>Experiment</h1>
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<p>In this experiment we succeed to stabilize the amount of living bacteria at a level of $0.015 OD_{600}$. Even better: the previsions gave us the right amount of bacteria. </p>
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<p>In the first part of the experiment, we first determined the parameters of the model that describe the behavior of our cell culture, according to the <a href="https://2013.igem.org/Team:Grenoble-EMSE-LSU/Project/Modelling/Density#MCBP>procedure described in the modeling part</a>. </p>
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<p>The first part of the experiment followed a protocol close to the other:</p>
<p>The first part of the experiment followed a protocol close to the other:</p>
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<p> <b>1</b>. A first period of 90 minutes in the dark for the bacteria to grow and produce KillerRed. </p>
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<p><b>1</b>.0-90 min : this first incubation was done in the dark to let the bacteria grow and produce KillerRed. </p>
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<p> <b>2</b>. A long period of 4 hours at an illumination of maximal intensity. At this time, the effect of illumination were clear, and some parameters could have been adapted to fit these effects.</p>
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<p><b>2</b>. 90-330 min : a longer 4 hours period with illumination at maximal intensity. After that time, the effects of light were clear, and the parameters of the model were calculated to fit the observable kinetics.</p>
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<p> Then the model was used to predict which illumination could stabilized the population.</p>  
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<p><b>3</b>. Following the model predictions, the illumination of the next period of timehas been at 70% of maximal intensity.</p>
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<p><b>3</b>. 330-450 min : Then our model was used to calculate the light intensity that would stabilize the population of living cells : light was applied at 70% of its maximal intensity.</p>
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<p><b>4</b>. After 2 hours, as the fluorescence and the absorbance were slowly drifting away from the predictions, the illumination was put at 50% of maximal intensity for 2 hours, and then at 40% of maximal intensity. </p>
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<p><b>4</b>. 450-720 min : After 2 hours, we noticed that the fluorescence and the absorbance were slowly drifting away from the predictions. Since the fluorescence was higher than expected and the OD was lower than expected, we reduced the illumination to 50% of its maximal intensity for 2 hours and then at 40% of its maximal intensity. </p>
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Revision as of 14:45, 3 October 2013

Grenoble-EMSE-LSU, iGEM


Grenoble-EMSE-LSU, iGEM

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