Difference between revisions of "Team:UGent Belgium/Demonstrate"

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<h1>Demonstration</h1>
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<br>
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<p>The goal of our project is to provide a working product that can collect a substantial amount of water. To this end, we performed some experiments to measure the exact quantity of water collected by our 3D-printed shape, coated with polylactic acid (PLA) and biotin, and treated with the Ice Nucleating Protein (INP)-streptavidin complex. A mixture of PLA and biotin, dissolved in dichloromethane, was used to coat the water collectors. The coated objects were left to dry overnight in a laminar flow cabinet, allowing the solvent to evaporate.</p>
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<p>We considered four biological treatments:</p>
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<ol style="list-style-type: decimal">
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<li><strong>INP + INP_NC-mSA2</strong>: whole cells expressing both full length INP and membrane-bound monomeric streptavidin (mSA2)</li>
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<li><strong>RFP + INP_NC-mSA2</strong>: Control, cells expressing membrane-bound mSA2 and a red fluorescent protein</li>
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<li><strong>INP_RC-mSA2</strong>: protein extract: INP nucleating domain - mSA2 fusion protein</li>
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<li><strong>mGFPuv2-mSA2</strong>: Control, mGFPuv2-mSA2 fusion protein</li>
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</ol>
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<p>In each experiment, the measurements were performed in a <a href="https://2016.igem.org/Team:UGent_Belgium/Hardware">controlled humidified chamber</a>. Both the temperature and the humidity were constantly measured and the latter was actively controlled using <a href="https://2016.igem.org/Team:UGent_Belgium/Software">bang-bang control</a>.</p>
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<h2 id="effect-biological-treatments-on-the-water-collectors">Effect biological treatments on the water collectors</h2>
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<h3 id="setup">Setup</h3>
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<p>In this demonstrative experiment, we assesed the effect of the four biological treatments described above.</p>
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<p>The dome of four water collectors were coated using the PLA-biotin solution. Subsequently each collector was sawn in half in order to have a treatment and associated control for each experimental unit.</p>
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<div class="center">
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<div class="figure">
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<img src="https://static.igem.org/mediawiki/2016/f/fb/T--UGent_Belgium--halfcollector.jpeg" alt="Half a water collector." width="600" />
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<p class="center"><i>Half of a water collector.</i></p>
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</div></div>
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<p>One half of each collector was treated with a different biological function (i.e. INP + INP_NC-mSA2, RFP + INP_NC-mSA2, INP_RC-mSA2 and mGFPuv2-mSA2). After waiting five minutes, to allow biotin-streptavidin bonds to form, all half-collectors were rinsed thoroughly with a physiological solution. Each halve was placed on a petri dish.</p>
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<p>All halves were dried and weighted on an analytical scale. Finally, all petri dishes were randomly placed in the humidified chamber. They were left under a flow of humid air for one night. Afterwards, all halves were weighed again, to measure the accumulation of water.</p>
  
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<img src="https://static.igem.org/mediawiki/2016/9/9c/T--UGent_Belgium--humchamb.jpeg" alt="The collectors in the humidified chamber." width="600"/>
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<p class="center"><i>The collectors in the humidified chamber.</i></p>
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</div></div>
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<h3 id="results">Results</h3>
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<p>The following bar chart shows the differences in water accumulation of each treatment-half compared to the control-half.</p>
  
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<p>Here you can describe the results of your project and your future plans. </p>
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<img src="https://static.igem.org/mediawiki/2016/4/41/Water_collection_collectors.png" alt="Difference in water collection treatment versus control for the four treatments." width="600"/>
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<p class="center"><i>Differences in water collection between treatment and control, for all four treatments.</i></p>
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<h5>What should this page contain?</h5>
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<p>The average water amount of water collected by all the experimental units is <span class="math">3.07 ± 0.38</span> gram. The difference in weight of the treatment-halves compared to the control-halves of the different treatments were between -0.92 and 0.6 gram. Unfortunately, it appears that the INP treatments result in a decrease in water collection. As the effect is quite small and we could not perform any replications, it is perhaps safest to conclude that these results do not support any conclusions either way.</p>
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<li> Clearly and objectively describe the results of your work.</li>
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<li> Future plans for the project </li>
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<li> Considerations for replicating the experiments </li>
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<h5> Project Achievements </h5>
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<script type="text/javascript" src="https://ajax.googleapis.com/ajax/libs/jquery/1.12.4/jquery.min.js"></script>
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<p>You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.</p>
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<li>A list of linked bullet points of the successful results during your project</li>
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<li>A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.</li>
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<h5>Inspiration</h5>
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<p>See how other teams presented their results.</p>
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<li><a href="https://2014.igem.org/Team:TU_Darmstadt/Results/Pathway">2014 TU Darmstadt </a></li>
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<li><a href="https://2014.igem.org/Team:Imperial/Results">2014 Imperial </a></li>
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<li><a href="https://2014.igem.org/Team:Paris_Bettencourt/Results">2014 Paris Bettencourt </a></li>
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Latest revision as of 18:46, 19 October 2016

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Demonstration


The goal of our project is to provide a working product that can collect a substantial amount of water. To this end, we performed some experiments to measure the exact quantity of water collected by our 3D-printed shape, coated with polylactic acid (PLA) and biotin, and treated with the Ice Nucleating Protein (INP)-streptavidin complex. A mixture of PLA and biotin, dissolved in dichloromethane, was used to coat the water collectors. The coated objects were left to dry overnight in a laminar flow cabinet, allowing the solvent to evaporate.

We considered four biological treatments:

  1. INP + INP_NC-mSA2: whole cells expressing both full length INP and membrane-bound monomeric streptavidin (mSA2)
  2. RFP + INP_NC-mSA2: Control, cells expressing membrane-bound mSA2 and a red fluorescent protein
  3. INP_RC-mSA2: protein extract: INP nucleating domain - mSA2 fusion protein
  4. mGFPuv2-mSA2: Control, mGFPuv2-mSA2 fusion protein

In each experiment, the measurements were performed in a controlled humidified chamber. Both the temperature and the humidity were constantly measured and the latter was actively controlled using bang-bang control.

Effect biological treatments on the water collectors

Setup

In this demonstrative experiment, we assesed the effect of the four biological treatments described above.

The dome of four water collectors were coated using the PLA-biotin solution. Subsequently each collector was sawn in half in order to have a treatment and associated control for each experimental unit.

Half a water collector.

Half of a water collector.

One half of each collector was treated with a different biological function (i.e. INP + INP_NC-mSA2, RFP + INP_NC-mSA2, INP_RC-mSA2 and mGFPuv2-mSA2). After waiting five minutes, to allow biotin-streptavidin bonds to form, all half-collectors were rinsed thoroughly with a physiological solution. Each halve was placed on a petri dish.

All halves were dried and weighted on an analytical scale. Finally, all petri dishes were randomly placed in the humidified chamber. They were left under a flow of humid air for one night. Afterwards, all halves were weighed again, to measure the accumulation of water.

The collectors in the humidified chamber.

The collectors in the humidified chamber.

Results

The following bar chart shows the differences in water accumulation of each treatment-half compared to the control-half.

Difference in water collection treatment versus control for the four treatments.

Differences in water collection between treatment and control, for all four treatments.

The average water amount of water collected by all the experimental units is 3.07 ± 0.38 gram. The difference in weight of the treatment-halves compared to the control-halves of the different treatments were between -0.92 and 0.6 gram. Unfortunately, it appears that the INP treatments result in a decrease in water collection. As the effect is quite small and we could not perform any replications, it is perhaps safest to conclude that these results do not support any conclusions either way.