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

 
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                         ">Imperial College London</a>
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                         ">Lab safety</a>
 
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                         ">Universidad Complutense de Madrid</a>
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                         ">Safety design in Printeria</a>
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                        ">Ethics in Printeria</a>
 
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             <h3>Introduction</h3>
 
             <h3>Introduction</h3>
 
          
 
          
             <p>As Theodor Reik (Sigmund Freud disciple) said: “In science, like in all life fields, a slogan is imposed: security first”. Nowadays, safety is an essential requirement when creating new devices that humans will use, and Printeria is not different.</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>
  
             <p>On the one hand biosafety describes  the containment principles. technologies and practices that are implemented to prevent the unintentional exposure to pathogens and toxines, or their accidental release [1]. On the other hand biosecurity describes de protection, control and accountability for valuable biological materials within laboratories, inorder to prevent their unauthorized access, loss, theft, misuse, diversion or intentional release.</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>
  
             <p>Next paragraphs describe how and why we implemented biosafety and biosecurity requirements in Printeria.</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;">Lab safety</p>
 
                   <p style="text-align: center !important;font-size: 1em !important;font-weight: bold;">Lab safety</p>
 
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                   <p style="text-align: center !important;font-size: 1em !important;font-weight: bold;">Safety design in Printeria</p>
 
                   <p style="text-align: center !important;font-size: 1em !important;font-weight: bold;">Safety design in Printeria</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>Lab Safety</h3>
 
                   <h3>Lab Safety</h3>
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                  <h4>Our laboratory</h4>
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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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      <h6>
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        <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.
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<p></p>
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                  <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>
 
                    
 
                    
                  <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>PixCell aims to get electronics and Synthetic Biology closer designing an innovative sistem where the culture is controlled by voltage.
 
                    &nbsp;</p>
 
 
                    
 
                    
                   <p> Antes de poner en marcha el gran proyecto que es Printeria, tenemos que comprobar que las construcciones genéticas y los métodos de laboratorio que vamos a utilizar, funcionan en un entorno controlado y seguro como es el de un laboratorio.
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                  <h4>Access to the laboratory</h4>
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                   <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>
  
Printeria utiliza como chasis bacterias E.coli de riesgo biológico 1 provenientes de las cepas DB3.1, 10G y TOP10.  Para las construcciones utilizamos biobricks y partes suministradas por IDT en Golden Gate.  Este trabajo, es realizado solamente por personas con experiencia en el campo y con conocimiento sobre la seguridad del laboratorio, como es el caso de los Biotecnólogos y los Ingenieros Biomédicos.
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                  <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>
  
Protocolos seguridad de antemano escrita y guiada por los instructors Alejandro vignoni y Yadira Boada Acosta
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                    <span class="btn__text">EVACUATION</span>
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                    <span class="btn__text">RIGHTS &amp; LIABILITIES</span>
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El laboratorio que utilizamos sigue la legislación vigente sobre el riesgo 1 (Ley 31/1995, RD 664/1997)  y tiene para ello un autoclave () y una campana () con esterilización mediante UV.  Además los resíduos generados por el laboratorio están minuciosamente controlados por el CGROM (centro generador de resíduos en la Oficina de Medioambiente) que facilitan los contenedores e intrucciones necesarias para la correcta clasificación de los resíduos.
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<p></p>
&nbsp;</p>
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                  <h4>Strains</h4>
 
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                  <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>
<p>Como medida de seguridad adicional, no se utiliza Bromuro de Etidio sino SYBR Safe, que tiene el mismo efecto sin efectos cancerígenos pero su uso es el mismo que con el BrEt en cuanto a protocolo.</p>
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                   <h3>Safety design in Printeria</h3>
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                   <h3>Safety by design</h3>
                   <p>Paris-Bettencourt team was right when they said : “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><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>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>Parameters</p></th>
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                                 <th><p><b>Safety Problem</b></p></th>
                                 <th><p>Description</p></th>  
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                                 <th><p style="text-align: center"> Implemented solution</p></th>  
 
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                                 <td><p>Number of samples</p></td>
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                                 <td><p><b>Sterilization of the PCB</b></p></td>
                                 <td><p>From <b>3 to 6 samples of reporter protein</b> and <b>3 samples of blank</b></p></td>  
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                                 <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>  
 
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                                 <td><p>Temperature</p></td>
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                                 <td><p><b>Sterilization of the microfluidic tubes</b></p></td>
                                 <td><p>37 ºC</p></td>  
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                                 <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>  
 
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                                 <td><p>Shake</p></td>
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                                 <td><p><b>Protection of the user</b></p></td>
                                 <td><p>Double Orbital. 01:00 (MM:SS)</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>
 
                               <tr>
 
                               <tr>
                                 <td><p>Absorbance. Optical Density (OD)</p></td>
+
                                 <td><p><b>Overheating of the PCB</b></p></td>
                                 <td><p>Wavelenght at <b>600 nm</b> emission</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>
                              <tr>
+
                              <tr>
                                 <td><p>Excitacion and emission scans</p></td>
+
                                 <td><p><b>Ventilation</b></p></td>
                                 <td><p>The scans occur between <b>two wavelength limit values</b>. The established range will depend on the theoretical spectrum of the protein.</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>
 
                               <tr>
 
                               <tr>
                                 <td><p>Excitacion and emission wavelengths</p></td>
+
                                 <td><p><b>Electroporator</b></p></td>
                                 <td><p>These values will depend on the range, and therefore on the spectrum. Values far from the theoretical peak lead to more attenuated fluorescence curves, and values very close to the peak can lead to overlap and error in reading the data. Therefore, <b>a compromise must be reached between curve resolution and reading overlap</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>
 
                               </tr>
                               <tr>
+
                                
                                <td><p>Gain (G)</p></td>
+
                            </tbody></table>
                                <td><p>Normally the gain value <b>G = 60</b>, although for proteins with lower fluorescence, it is recommended that G takes higher values.</p></td>
+
                         <p></p>
                              </tr>
+
                            </table>
+
                         </p></li>
+
  
                <p>Lista + Comité de ética.
 
Riesgo y posibles soluciones.
 
</p>
 
  
 
                  
 
                  
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 +
 +
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 +
 +
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                <p></p>
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               </div>
 
               </div>
 
             </div>
 
             </div>
         </div>
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<a class="anchorOffset" id="Safety"></a>
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                  <img src="https://static.igem.org/mediawiki/2018/c/c3/T--Valencia_UPV--Etica_imagenUPV2018.png" style="
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                  <p></p>
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              </div>
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              <div class="col-md-9">
 +
                  <h3>Ethics in Printeria</h3>
 +
<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></p>
 +
 
 +
<div style="
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                    text-align: center;"><a class="btn btn--primary-1" href="https://static.igem.org/mediawiki/2018/6/63/T--Valencia_UPV--Resolucion_eticaUPV2018.pdf" style="
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                    <span class="btn__text">COMMITTEE VEREDICT</span>
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                    </a>
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                  </div>
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              </div>
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            </div>
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        </div></div>
  
 
       </div>
 
       </div>
 
   </div>
 
   </div>
  
 +
        <div>
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            <h3>References</h3>
 +
       
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            <p itemscope="" itemtype="http://schema.org/Book">
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    <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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</p>
  
 +
        </div>
 
</div>
 
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</section>
 
</section>
  

Latest revision as of 19:56, 17 October 2018

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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.

References

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

CONTACT US igem.upv.2018@gmail.com