Difference between revisions of "Team:Valencia UPV/Safety"

 
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               ">Index</h4>
 
               ">Index</h4>
 
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                        ">Harvard University</a>
 
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                         ">Universidad de las fuerzas armadas ESPE</a>
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                         ">Lab safety</a>
 
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                         ">Philipps-Universität Marburg</a>
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                         ">Safety design in Printeria</a>
 
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                        ">Imperial College London</a>
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                         ">Universidad Complutense de Madrid</a>
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                         ">Ethics in Printeria</a>
 
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             <h3>Introduction</h3>
 
             <h3>Introduction</h3>
 
          
 
          
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             <p>As a team, one of our favourite parts in iGEM have been collaborations. They reminded us that iGEM is not only a competition but a chance to share and let other teams be a part of your project, and that is wonderful. Below is shown the different universities that have collaborated with us. By clicking in the index or in the images you can go directly to the explanation.</p>
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             <p>As Theodor Reik (Sigmund Freud disciple) said: “In science, like in all life fields, a slogan is imposed: <b>safety first</b>”. Nowadays, safety is an essential requirement when creating new devices which humans will use, and Printeria is not different.</p>
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            <p>This document describes the safety of our project so, a clever step before begining is to know what is it. Safety describes  the containment principles, technologies and practices that are implemented to prevent the unintentional exposure to pathogens and toxines, or their accidental release [1]. Furthermore, safety is how you have protected yoursef and other in order to avoid any harms which could occur.</p>
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            <p><b>Printeria is designed to be a secure device</b>, because is going to be used by bioartists, schools and researchers which are our identified potential users according to <a href="https://static.igem.org/mediawiki/2018/3/32/T--Valencia_UPV--Potential_Users_MITUPV2018.pdf" target="blank">MIT segmentation table</a> and we do not want it to be a variable which create concern in those users. This approach toguether with our lab tools and posibilities,  have  influenced the experiments we did and the way we build Printeria. Next paragraphs describe our lab safety and how and why we implemented biosafety and biosecurity requirements in Printeria.</p>
  
 
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                  <p style="text-align: center !important;font-size: 1em !important;font-weight: bold;">Harvard University</p>
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                  <a class="inner-link" href="#fuerzasArmadas"><img class="fotosAttr" src="https://static.igem.org/mediawiki/2018/7/7b/T--Valencia_UPV--Icono_EcuadorUPV2018.png"></a>
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                  <p style="text-align: center !important;font-size: 1em !important;font-weight: bold;">Universidad de las Fuerzas Armadas ESPE</p>
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                  <p style="text-align: center !important;font-size: 1em !important;font-weight: bold;">Philipps-Universität Marburg</p>
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                   <p style="text-align: center !important;font-size: 1em !important;font-weight: bold;">Lab safety</p>
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                  <a class="inner-link" href="#complutense"><img class="fotosAttr" src="https://static.igem.org/mediawiki/2018/9/90/T--Valencia_UPV--Safety_DesignUPV2018.png"></a>
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                  <p style="text-align: center !important;font-size: 1em !important;font-weight: bold;">Safety design in Printeria</p>
 
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               </div>
 
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                   <a class="inner-link" href="#Safety"><img class="fotosAttr" src="https://static.igem.org/mediawiki/2018/c/c3/T--Valencia_UPV--Etica_imagenUPV2018.png"></a>
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                   <p style="text-align: center !important;font-size: 1em !important;font-weight: bold;">Universidad Complutense de Madrid</p>
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                   <p style="text-align: center !important;font-size: 1em !important;font-weight: bold;">Ethics in Printeria</p>
 
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                  <h3>Harvard</h3>
 
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                  <h4>Team Project</h4>
 
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                  <p>When explaining Printeria to people, we see they are reticent to use it because of the genetically modified bacteria that we do. A part of our project is to fight the stigma against Synthetic Biology and Harvard was claiming for teams disposed to do so.&nbsp;</p>
 
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                  <h4>Collaboration</h4>
 
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                  <p> Their collaboration was about doing a report of your project and create a series of newsletters and articles to show how we are using bacteria and demonstrate that we are doing it taking into account demanding security measures. However, throughout the summer they changed their mind and now they are doing a magazine instead of a series of articles. They will distribute it in their university and of course in the Giant Jamboree. You can download our report and the magazine by clicking in the buttons below.&nbsp;</p>
 
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                   <h3>Ecuador</h3>
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                   <h3>Lab Safety</h3>
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                   <h4>Team Project</h4>
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                   <h4>Our laboratory</h4>
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                   <p><a href="https://2018.igem.org/Team:Ecuador" target="blank">Ecuador iGEM team</a> has been working on the development of a new biomaterial to speed up the healing in bone injures. This biomaterial called (C-lastine), is based on the cross-linking of bacterial cellulose and Elastin-like polypeptides. This cross-linking is mediated by a carbohydrate binding domain and a BMP2 protein (an osteoblasts differentiation inducer). To achieve this objective they are using E.Coli as a biofactory, specifically C41 strain which is resistant to Cellulose synthase operon protein D, codified by gene bcsD and responsible of pore formation in the membrane allowing cellulose to exit the cell. The final result is a bandage matrix that can be applied to the wound during a surgical intervention.</p>
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                   <p>We have planned this project consequently with the tools we have. <b>Our lab follows the current spanish legislation</b> about risk 1 (Ley 31/1995, RD 664/1997) and as it is written, our lab is provided with an <b>autoclave</b> and an laminar <b>flow cabinet with UV</b> sterilization. Waste generated are minutely controlled by <b>CGROM</b> (waste management center of the Polytechnic University of Valencia). They provide every lab with special containers and  instructions for the correct classification of waste which can be seen in the next photos:</p>
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                   <h4>Collaboration</h4>
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                   <p> If we think about it, Printeria is a kind of biofactory so, we thought that trying to make Ecuadorís biomaterial in Printeria would be a good trial to show that Printeria is ready to become something greater in the biotechnological industry and why not, it could be used in hospitals as well. However, due to the fact that there were problems with the size of the parts for Golden Gate and with the customs, it could not become a reality. This idea would be developed with Imperial College. Instead, we did several things.</p>
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                   <p>We did a survey to get some feedback from the potential users of both projects. It consisted in a <a href="https://www.youtube.com/watch?v=ITVeAXSsO5k" style="padding-right: 0;" target="blank">video</a> in spanish and then a survey to fill out. In our case, the survey was made to know what users  thought about Printeria and Synthetic Biology. In no case this survey represents the collective of users (only 15 people were surveyed) it was just made to have an idea.</p>
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 +
      <h6>
 +
        <b>Images 1-2.</b> On the left, Roger explaining Marc how to deal with chemical containers, on the right, the containers which CGROM manage.
 +
      </h6>
 +
    </div>
 +
  </div>
 +
<p></p>
 +
                  <p>As an additional safety measure we use <b>SYBR Safe</b> instead of <b>Eidium bromide</b>. Its use is identical in all the protocols but without carcinogenic risks.</p>
 +
                 
 +
                 
 +
                   <h4>Access to the laboratory</h4>
 +
                   <p>As we are a multidisciplinary team formed by biotechnologists, engineers and artists, <b>not everybody knows how to behaviour in a laboratory</b>. That is why we did several <b>seminars</b> directed by Alejandro Vignoni and Yadira Boada to <b>teach the basics</b> of their laboratory. However, as a security measure, they decided that not everybody would be allowed to work in the lab alone, only biotechlogists and biomedical engineers could be there without surveillance due to their experience and knowledge adquiered during their degree.</p>
 +
 
 +
                   <p>Over the course of these seminars we were informed about the <b>evacuation</b> protocols, <b>waste management</b> and our <b>rights and liabilities</b> as UPV staff. These information can be seen in the information sheets they gave us and that you can download through these buttons:</p>
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 +
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 +
<p></p>
 +
                  <h4>Strains</h4>
 +
                  <p>Printeria uses <b><i>E.coli</i></b> and <b><i>V.natriegens</i></b> as chassis. Both are classified as <b>risk 1</b> so the harm which can cause to people is minimal. The <i>E.coli</i> are from strains DB3.1, 10G and TOP10, for <i>V.natriegens</i> we use the strain  ATCC 1404 from <b>CECT</b> (Colección Española de Cultivos Tipo). To do constructions we use  biobricks and parts supplied by IDT prepared with the Golden Gate grammar.</p>
  
  
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                  <p>Furthermore we provide them with a special strain called C41. This was really important for them due to the fact that without it, they could not continue with their iGEM project.</p>
 
 
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                   <h3>Marburg</h3>
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                   <h3>Safety by design</h3>
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                   <h4>Team Project</h4>
+
                   <p><b>Paris-Bettencourt 2012 team</b> was right when they said in their page "<i>How safe is safe enough?</i>": “Biosafety is an exciting design challenge, an essential enabling technology for synthetic biology, and a fundamental ethical obligation of all bioengineers”.</p>
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                  <p>Do you know that there is something that grows faster than a Bacteria? If it grows faster...why not use it as a chassis to do pharmaceuticals and chemicals in the industry? This is the fantastic idea that Marburg team has developed. They want to demonstrate that working with V.Natriegens is cheaper, faster and possible with todays methods.&nbsp;</p>
+
−
                  <h4>Collaboration</h4>
+
−
                  <p> Initially, they requested us to do the interlab study with V.natriegens and to annotate how fast V.natriegens grows but this gave us a really good idea. Why don't we try to use V.natriegens in Printeria? If all goes well, it would mean that Printeria is ready to work with other organisms which grow faster than E.coli, this way we reinforce the biofactory idea of Printeria.&nbsp;</p>
+
−
                  <p> However, as things do not go as we spect in certain moments we could not make the interlab with V.natriegens. (Aunque) Marburg team gave us almost all the elements to do it we had to buy V.natriegens in the CECT and unfortunately they arrived one week before the deadline. We tried several times using a (alquilada) frozen centrifuge but 3 tries later we could not achieve electrocompetent cells. As we do not have sponsors and (alquilando) the frozen centrifuge was not cheap we stoped trying. &nbsp;</p>
+
−
                  <p>on the other side, trying V.natriegens in Printeria has been an an enormous success. We achieved the (ensablaje) of (no se cuantas partes). You can see the results in the next photo:  &nbsp;</p>
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                  <p>This sentence describes perfectly what we did with Printeria.  Printeria was designed to ensure safety for all audiences, <b>to be safe enough for the world</b>. In the next table you can see <b>different safety problems we saw with our instructors help, and how we solved them.</b></p>
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 +
                                <th><p><b>Safety Problem</b></p></th>
 +
                                <th><p style="text-align: center"> Implemented solution</p></th>
 +
                              </tr>
 +
                              <tr>
 +
                                <td><p><b>Sterilization of the PCB</b></p></td>
 +
                                <td><p>How do we clean the PCB (Printed Circuit Board) without <b>damaging</b> metalic and plastic components?</p><p> Well, we decided to copy what flow cabins do. Based on this, we decided to use <b>UV led lights</b> to achieve this objective. This idea was given by <b>Mojica</b> as a part of our <a href="https://2018.igem.org/Team:Valencia_UPV/Integrated_Human" target="blank">intergrated human</a>. Furthermore we have programmed an especial reaction that use little droplets with <b>HCl and ethanol</b> to clean and ensure we have perfectly sterilized the PCB which is protected by a <b>plastic film</b> as well and this feature prevents acid from attacking it.</p></td>
 +
                              </tr>
 +
                              <tr>
 +
                                <td><p><b>Sterilization of the microfluidic tubes</b></p></td>
 +
                                <td><p>The reaction fluid is going to pass through <b>microfluidic tubes</b> and it will <b>contaminate them</b>. To reverse this situation we will use the same special program wich usses droplets with <b>ethanol and HCl</b>.</p></td>
 +
                              </tr>
 +
                              <tr>
 +
                                <td><p><b>Protection of the user</b></p></td>
 +
                                <td><p>We want users to see what is happening inside Printeria but also we want them to be protected if something wrong happens inside. That is why we thought that a <b>metacrilate box</b> that covered the machine would be a nice option. Besides, it is useful to <b>pin up the elements</b> which will be part of Printeria.</p></td>
 +
                              </tr>
 +
                              <tr>
 +
                                <td><p><b>Overheating of the PCB</b></p></td>
 +
                                <td><p>As the <b>PCB has a cold and a hot zone</b>, it is necessary to evacuate the heat generated. The hot zone can achieve more than 80º degrees and can be a potencial danger. A resistance is responsible of heating, and a <b>peltier plate</b> is responsible of chilling. Peltier effect is based in a semiconductor metal where you apply a potential difference. Then, you create a hot part and a cold one in that semiconductor. We use the cold part to extract the energy and stabilize the temperature via conduction of the PCB's hot zone.</p></td>
 +
                              </tr>
 +
                              <tr>
 +
                                <td><p><b>Ventilation</b></p></td>
 +
                                <td><p>The peltier plate is responsible of chilling the PCB but <b>how do we evacuate all the energy generated?</b> Well, the answer is simple, Printeria just needed <b>ventilation holes</b> and <b>watercooling</b> to do it. Watercooling is based in the higher termic capacity of fluids (water in this case) to absorb energy. Water is pumped through microfluidic tubes to reach the peltier and chill it.</p></td>
 +
                              </tr>
 +
                              <tr>
 +
                                <td><p><b>Electroporator</b></p></td>
 +
                                <td><p>This device is capable to reach high voltages, so if you require one of these in your design first of all, ensure no one can harm himself/herself. This is the reason why we designed our own electroporator in a <b>dedicated and compacted printed circuit to isolate it</b> from any exposure. It can reach until 1600V, which is not a joke.
 +
                              </p></td></tr><tr>
 +
                                <td><p><b>Tips</b></p></td>
 +
                                <td><p>Tips which go in the revolver wheel are <b>disposable</b> to ensure the purity of each reaction.</p></td>
 +
                              </tr>
 +
                             
 +
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 +
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                  <h3>Imperial College</h3>
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                  <h4>Team Project</h4>
+
−
                  <p>This collaboration was born when Ismael (Imperial College iGEM team supervisor) came to the Univertitat Politécnica de Valencia to have a meeting with Diego Orzáez one of our instructors. Taking advantage of that situation we met with him and he explained us the Imperial College project called PixCell.</p>
+
−
                  <p>PicCell aims to get electronics and Synthetic Biology closer designing an innovative sistem where the culture is controlled by voltage.
+
−
                    &nbsp;
+
−
                  </p>
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−
                  <h4>Collaboration</h4>
+
−
                  <p> As they are using Golden Gate grammar we thought that it could be a good opportunity to do what we could not do with Ecuador team, try their construction in Printeria and compare results handmade and Printeria made.&nbsp;</p>
+
 
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                   <h3>Complutense de Madrid</h3>
+
                   <h3>Ethics in Printeria</h3>
−
                  <h4>Collaboration</h4>
+
<p><b>Science without ethics is cruel, and ethics without science is empty</b>. Ethics become really important when a project is being thought. For this reason we wanted to pass the UPV (Polytechnic University of Valencia) ethics committee. Here you can download the document we sent them to approve and the <b>positive veredict</b> of the committee. </p>
−
                  <p> One of biggest problems during iGEM project is to learn how to work with Wiki sintaxis. As not everybody is acquainted with programming with marking languages like html or xml, managing the wiki is a difficult task for every team. &nbsp;</p>
+
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−
                  <p>Francisco Javier Quero, team leader of Madrid-OLM team, contacted us at the beginning of the summer to ask for help with their wiki. To help them solve their problems our software team did a call via Skype to talk these topics:</p>
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                  <ul style="list-style-type:square">
 
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                    <li><p>An explanation of how mediawiki works and the main problems they will find working with it.</p></li>
 
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 +
            <h3>References</h3>
 +
       
 +
            <p itemscope="" itemtype="http://schema.org/Book">
 +
    <cite itemprop="name"> <b>1.</b> Dual use: A report by iGEM team Bielefeld-CeBiTec 2015</cite>. [Germany]. <span itemprop="publisher">Universität Bielefeld</span>, <time itemprop="datePublished">2015</time>. <span itemprop="numberOfPages">30 pages.</span>
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Latest revision as of 19:56, 17 October 2018

Stack Multipurpose HTML Template

Index

Introduction

As Theodor Reik (Sigmund Freud disciple) said: “In science, like in all life fields, a slogan is imposed: safety first”. Nowadays, safety is an essential requirement when creating new devices which humans will use, and Printeria is not different.

This document describes the safety of our project so, a clever step before begining is to know what is it. Safety describes the containment principles, technologies and practices that are implemented to prevent the unintentional exposure to pathogens and toxines, or their accidental release [1]. Furthermore, safety is how you have protected yoursef and other in order to avoid any harms which could occur.

Printeria is designed to be a secure device, because is going to be used by bioartists, schools and researchers which are our identified potential users according to MIT segmentation table and we do not want it to be a variable which create concern in those users. This approach toguether with our lab tools and posibilities, have influenced the experiments we did and the way we build Printeria. Next paragraphs describe our lab safety and how and why we implemented biosafety and biosecurity requirements in Printeria.

Lab safety

Safety design in Printeria

Ethics in Printeria

Lab Safety

Our laboratory

We have planned this project consequently with the tools we have. Our lab follows the current spanish legislation about risk 1 (Ley 31/1995, RD 664/1997) and as it is written, our lab is provided with an autoclave and an laminar flow cabinet with UV sterilization. Waste generated are minutely controlled by CGROM (waste management center of the Polytechnic University of Valencia). They provide every lab with special containers and instructions for the correct classification of waste which can be seen in the next photos:

Images 1-2. On the left, Roger explaining Marc how to deal with chemical containers, on the right, the containers which CGROM manage.

As an additional safety measure we use SYBR Safe instead of Eidium bromide. Its use is identical in all the protocols but without carcinogenic risks.

Access to the laboratory

As we are a multidisciplinary team formed by biotechnologists, engineers and artists, not everybody knows how to behaviour in a laboratory. That is why we did several seminars directed by Alejandro Vignoni and Yadira Boada to teach the basics of their laboratory. However, as a security measure, they decided that not everybody would be allowed to work in the lab alone, only biotechlogists and biomedical engineers could be there without surveillance due to their experience and knowledge adquiered during their degree.

Over the course of these seminars we were informed about the evacuation protocols, waste management and our rights and liabilities as UPV staff. These information can be seen in the information sheets they gave us and that you can download through these buttons:

Strains

Printeria uses E.coli and V.natriegens as chassis. Both are classified as risk 1 so the harm which can cause to people is minimal. The E.coli are from strains DB3.1, 10G and TOP10, for V.natriegens we use the strain ATCC 1404 from CECT (Colección Española de Cultivos Tipo). To do constructions we use biobricks and parts supplied by IDT prepared with the Golden Gate grammar.

Safety by design

Paris-Bettencourt 2012 team was right when they said in their page "How safe is safe enough?": “Biosafety is an exciting design challenge, an essential enabling technology for synthetic biology, and a fundamental ethical obligation of all bioengineers”.

This sentence describes perfectly what we did with Printeria. Printeria was designed to ensure safety for all audiences, to be safe enough for the world. In the next table you can see different safety problems we saw with our instructors help, and how we solved them.

Safety Problem

Implemented solution

Sterilization of the PCB

How do we clean the PCB (Printed Circuit Board) without damaging metalic and plastic components?

Well, we decided to copy what flow cabins do. Based on this, we decided to use UV led lights to achieve this objective. This idea was given by Mojica as a part of our intergrated human. Furthermore we have programmed an especial reaction that use little droplets with HCl and ethanol to clean and ensure we have perfectly sterilized the PCB which is protected by a plastic film as well and this feature prevents acid from attacking it.

Sterilization of the microfluidic tubes

The reaction fluid is going to pass through microfluidic tubes and it will contaminate them. To reverse this situation we will use the same special program wich usses droplets with ethanol and HCl.

Protection of the user

We want users to see what is happening inside Printeria but also we want them to be protected if something wrong happens inside. That is why we thought that a metacrilate box that covered the machine would be a nice option. Besides, it is useful to pin up the elements which will be part of Printeria.

Overheating of the PCB

As the PCB has a cold and a hot zone, it is necessary to evacuate the heat generated. The hot zone can achieve more than 80º degrees and can be a potencial danger. A resistance is responsible of heating, and a peltier plate is responsible of chilling. Peltier effect is based in a semiconductor metal where you apply a potential difference. Then, you create a hot part and a cold one in that semiconductor. We use the cold part to extract the energy and stabilize the temperature via conduction of the PCB's hot zone.

Ventilation

The peltier plate is responsible of chilling the PCB but how do we evacuate all the energy generated? Well, the answer is simple, Printeria just needed ventilation holes and watercooling to do it. Watercooling is based in the higher termic capacity of fluids (water in this case) to absorb energy. Water is pumped through microfluidic tubes to reach the peltier and chill it.

Electroporator

This device is capable to reach high voltages, so if you require one of these in your design first of all, ensure no one can harm himself/herself. This is the reason why we designed our own electroporator in a dedicated and compacted printed circuit to isolate it from any exposure. It can reach until 1600V, which is not a joke.

Tips

Tips which go in the revolver wheel are disposable to ensure the purity of each reaction.

Ethics in Printeria

Science without ethics is cruel, and ethics without science is empty. Ethics become really important when a project is being thought. For this reason we wanted to pass the UPV (Polytechnic University of Valencia) ethics committee. Here you can download the document we sent them to approve and the positive veredict of the committee.

References

1. Dual use: A report by iGEM team Bielefeld-CeBiTec 2015. [Germany]. Universität Bielefeld, . 30 pages.

CONTACT US igem.upv.2018@gmail.com