Team:INSA Toulouse/contenu/safety/safety in the lab

From 2013.igem.org

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   <h1 class="title1">Safety in the Lab</h1>
   <h1 class="title1">Safety in the Lab</h1>
-
   <h2 class="title2">Concerning our institution, safety in INSA Toulouse</h2>
+
   <h2 class="title2">Safety in INSA Toulouse</h2>
-
   <p class="texte">INSA Toulouse is a public school, which have some laboratories used for public research. Everybody in the INSA must also follow the French word code, which legislates for respect of environment, workers and public safety. Most of the rules we have to follow are presented on the manual “<a href="http://www.who.int/csr/resources/publications/biosafety/LabBiosMan3rdFrenchweb.pdf">Manuel de sécurité en laboratories</a>” (from the WHO), which permit us to know almost everything we have to know about safety with microorganisms we use and products we manipulate. In addition, daily use of Deming wheel allows us to improve the safety system and keep it updated.</p>
+
   <p class="texte">INSA Toulouse is a public engineer school in which several government research laboratories are present. Workers at INSA must follow the French rules concerning safety and environmental regulations. Most of these rules are presented in the manual “<a href="http://www.who.int/csr/resources/publications/biosafety/LabBiosMan3rdFrenchweb.pdf" target="_blank">Manuel de sécurité en laboratoires</a>” (from the WHO), which allowed us to understand and apply safety rules when manipulating microorganisms and chemicals. In addition, daily use of the Deming wheel allowed us to improve the safety rules and keep them updated.</p>
    
    
   <h2 class="title2">Concerning the laboratory</h2>
   <h2 class="title2">Concerning the laboratory</h2>
 +
<h3 class="title3left530">Personal protective equipment</h3>
 +
 +
  <p class="texteleft530">
 +
We used conventional protection all along our daily manipulating. All experiments were perfomed with gloves and a conventional lab coat, protecting the worker and avoiding contaminations. We also wear glasses when needed (UV exposition, hot water manipulations, chemicals manipulation, etc.).</p>
 +
 +
<img src="https://static.igem.org/mediawiki/2013/2/2b/Hang.png" class="imgcontentright" />
 +
<div class="clear"></div>
 +
 +
 +
 
 +
<h3 class="title3">Biological and Chemical Waste</h3>
 +
  <p class="texteleft480">After a 1h30 training for chemical waste gesture and biological hazard, we learned how to use different trash containers as described in the picture. Biological waste, potentially containing viable microorganism, were autoclaved in a special bag, then discarded. Chemical wastes are grouped by categories and eliminated via the regular procedure for all chemicals of the INSA.</p>
 +
 +
 +
<img src="https://static.igem.org/mediawiki/2013/7/70/Waste_-_200PX.png" class="imgcontentright" />
 +
<div class="clear"></div>
 +
 +
 +
  <h3 class="title3">Devices and Material</h3>
 +
 +
  <p class="texte">Here is a list of devices we used and how to use them safely:</p>
 +
 +
 +
 +
 
  <div class="list">
  <div class="list">
      
      
     <ul class="circlearrow">
     <ul class="circlearrow">
-
       <li> <span class="title3">Worker protections</span></br>
+
       <li> <span class="title3">Chemical Storage</span></br>
-
<span  class="texte">As in every lab, we use conventional protection when manipulating. Every experiment has to be made with gloves and a conventional lab coat , protecting the worker and avoiding contaminations. We also wear glasses when they are useful (UV exposition, EtBr using, hot water manipulations, chemicals manipulation…).</span></li>
+
<span  class="texte">We got separated and dedicated racks for every kind of chemical product we use. As you can see on these pictures, those boxes are key-closed.</span></li>
-
<br>
+
</ul>
-
       <li> <span class="title3">Waste gesture</span></br>
+
  </div>
-
<span class="texte">After a formation of chemical waste gesture and biological hazard, we learn how to use different trash cabs as described in the picture.  
+
 
-
Biological wastes, potentially containing viable microorganism, are firstable autoclaved in a special bag, then discard on the classical way.
+
<img src="https://static.igem.org/mediawiki/2013/7/77/Inflammable_-_75px.png" class="imgcontentleft" />
-
Chemical waste are regrouped and treated every month together with wastes of other labs of the campus.</span></li>
+
<img src="https://static.igem.org/mediawiki/2013/c/cb/Acides_-_75px.png" class="imgcontentleft" />
-
      <li>Cras vitae felis at lacus ele </li>
+
<img src="https://static.igem.org/mediawiki/2013/0/06/Bases_-_75px.png" class="imgcontentleft" />
-
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+
 
-
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+
<div class="clear"></div>
 +
<div class="list">
 +
   
 +
    <ul class="circlearrow">
 +
      <li> <span class="title3">Ethidium bromide</span></br>
 +
<span class="texte">A dark room is dedicated for using EtBr and UV. This room is key-closed. Everyone entering the room must wear gloves, glasses and lab coat. Two special boxes are available to discard spoiled EtBr (one for used agarose gels, a second for gloves).</span></li>
 +
<img src="https://static.igem.org/mediawiki/2013/9/90/EtBr_wastes_-_400_px.png" class="imgcontentleftmarge" />
 +
</ul>
 +
 
 +
  </div>
 +
 
 +
<img src="https://static.igem.org/mediawiki/2013/7/7a/EtBR_room.png" class="imgcontentleft" />
 +
 
 +
 
 +
 
 +
 
 +
<div class="clear"></div>
 +
 
 +
<div class="list">
 +
   
 +
    <ul class="circlearrow">
 +
       <li> <span class="title3">Biological safety cabinet</span></br>
 +
<span class="texte">To avoid external contamination by unwanted microorganisms on agar plates we used a Biological safety cabinet (FASTER – Ultrasafe). The cabinet is cleaned-up every morning when we arrive and every evening when we leave.</span></li>
 +
 
 +
</ul>
 +
 
 +
  </div>
 +
 
 +
<img src="https://static.igem.org/mediawiki/2013/1/19/PSM_-_180px.png" class="imgcontentleft" />
 +
 
 +
 
 +
 
 +
 
 +
<div class="clear"></div>
 +
 
 +
<div class="list">
 +
   
 +
    <ul class="circlearrow">
 +
      <li> <span class="title3">Electric burner</span></br>
 +
<span class="texte">For all sterile manipulations, we used electric burners. They are as efficient as Bunsen burners (even more), but much safer than them (limiting greatly the risks of fire hazards).</span></li>
 +
</ul>
 +
 
 +
  </div>
 +
 
 +
<img src="https://static.igem.org/mediawiki/2013/b/b8/Elec_burner.png" class="imgcontentleft" />
 +
 
 +
<div class="clear"></div>
 +
 
 +
<div class="list">
 +
   
 +
    <ul class="circlearrow">
 +
       <li> <span class="title3">Chemical Hood</span></br>
 +
<span class="texte">To protect the user from volatile chemical compounds while dangerous manipulations, we used of two chemical hoods.</span></li>
 +
</ul>
 +
 
 +
  </div>
 +
 
 +
<img src="https://static.igem.org/mediawiki/2013/8/86/Fume_Cupboard.png" class="imgcontentleft" />
 +
 
 +
<div class="clear"></div>
 +
 
 +
<div class="list">
 +
   
 +
    <ul class="circlearrow">
 +
       <li> <span class="title3">Water-bathes</span></br>
 +
<span class="texte">Use of water-bathes can be dangerous, with exposition of boiling or hot water. We used special gloves for protecting us from projections and steam. The use of water-bathes implies of course wearing glasses and lab coat. Special care was taken to switch them off every evening before leaving!</span></li>
 +
 
     </ul>
     </ul>
   </div>
   </div>
 +
 +
<img src="https://static.igem.org/mediawiki/2013/5/5c/Hot_bathe.png" class="imgcontentleft" />
<div class="clear"></div>
<div class="clear"></div>
-
  <h3 class="title3">Worker protections</h3>
+
<div class="list">
 +
   
 +
    <ul class="circlearrow">
 +
      <li> <span class="title3">P2 Laboratory</span></br>
 +
<span class="texte">Special care have been taken to manipulate the <i>Chromobacterium violaceum</i> strain which is classified as a class 2 microorganism. All manipulations with this bacteria were performed in a a dedicated P2 laboratory, fully equipped for microbiology. Biological waste from C. violaceum were treated with special procedures according to the Safety Regulation Rules for Class II microorganisms. </span></li>
-
  <p class="texte">As in every lab, we use conventional protection when manipulating. Every experiment has to be made with gloves and a conventional lab coat , protecting the worker and avoiding contaminations. We also wear glasses when they are useful (UV exposition, EtBr using, hot water manipulations, chemicals manipulation…).</p>
+
    </ul>
-
  <h3 class="title3">Waste gesture</h3>
+
  </div>
-
  <p class="texte">After a formation of chemical waste gesture and biological hazard, we learn how to use different trash cabs as described in the picture.
+
<img src="https://static.igem.org/mediawiki/2013/1/19/P2_lab.png" class="imgcontentleft" />
-
Biological wastes, potentially containing viable microorganism, are firstable autoclaved in a special bag, then discard on the classical way.
+
<div class="clear"></div>
-
Chemical waste are regrouped and treated every month together with wastes of other labs of the campus.</p>
+
-
 
+
-
   <p class="texteleft"><span class="spantitle">lorem ipsum</span></br>
+
   <h2 class="title2">Concerning our <i>E. calculus</i> project</h2>
-
<br>Mauris ac suscipit erat, sit amet blandit lectus. Fusce placerat, lectus at suscipit viverra, lorem lorem mattis turpis, id pharetra velit nunc vitae velit. Etiam ultrices aliquam ligula, sed ultrices nibh vulputate non. Ut dapibus arcu luctus, suscipit urna et, imperdiet magna. Curabitur tristique sed diam non elementum. Sed lectus urna, consequat quis porta eu, ultrices in nunc. Aenean vitae elit neque.<p>
+
-
  <p class="texteright"><span class="spantitle">lorem ipsum</span></br>
+
  <h2 class="title2">Would <i>E. calculus</i> raise safety issues in terms of:</h2>
-
<br>Mauris ac suscipit erat, sit amet blandit lectus. Fusce placerat, lectus at suscipit viverra, lorem lorem mattis turpis, id pharetra velit nunc vitae velit. Etiam ultrices aliquam ligula, sed ultrices nibh vulputate non. Ut dapibus arcu luctus, suscipit urna et, imperdiet magna. Curabitur tristique sed diam non elementum. Sed lectus urna, consequat quis porta eu, ultrices in nunc. Aenean vitae elit neque.</p>
+
-
<div class="clear"></div>
 
-
    
+
<div class="list2">
 +
   
 +
    <ul class="circlearrow">
 +
      <li> <span class="title3">Researcher safety?</span></br>
 +
<span  class="texte">In this project, we are working with biosafety level 1 microorganism (according to World Health Organization (WHO) <a href="http://www.who.int/csr/resources/publications/biosafety/Biosafety7.pdf" target="_blank"> (Laboratory Biosafety Manual</a>) with Escherichia coli DH5-alpha, DH5-1 and XL1-Blue strains, which are considered as biologically safe for users. Some of chemical compounds are dangerous for health (EtBr, Phenolchloroform, Chloramphenicol), but as previously described, all precautions are taken to ensure security and safety for all users.</span></li>
 +
<br>
 +
      <li> <span class="title3">Public safety?</span></br>
 +
<span class="texte">As described in the overview, <i>E. calculus</i> is not designed to be used on other environments than the laboratory. Interactions with human body are not envisaged and public should also never be in contact with our modified organism. All the experiments we have performed we made according to the highest standards of safety measures. Each modification we apply to our strains should not be threatening for biodiversity and external organisms beacuse the <i>E. coli</i> strains that are routinely used in the laboratories are mutated in many genes, and should not be able to survive in an external environment. <br>
 +
However, we are conscious that the gates we designed could potentially (and hopfully :-)) used for other biological applications, especially in metabolic engineering. In our view, these potential applications will probably be part of a process in a closed reactors, or in controlled environment, hence the safety regulations in use in the industrial context will apply. We also believe that our system present no evident potential danger, mainly because the used genes or parts were never described as potential threats to human and hopefully will never be an issue for public safety.
 +
</span></li>
 +
<br>
 +
      <li> <span class="title3">Environmental safety?</span></br>
 +
<span class="texte"><i>E. calculus</i> should  not be disseminated in the environment of course. Avoiding contaminations and accidental release of the <i>E. coli</i> strain implies good laboratory practise, particularly for elimination of biological waste (see above).</span></li>
 +
    </ul>
 +
 
 +
   </div>
 +
 
<div class="clear"></div>
<div class="clear"></div>
-
  <h3 class="title3">TITRE 3 lorem ipsum</h3>
 
-
   <p class="texte">Lorem ipsum dolor sit amet, consectetur adipiscing elit. Ut et dolor turpis. Suspendisse interdum, dolor eu  ultricies dignissim, arcu sapien placerat quam, ut bibendum lorem nibh id magna. Nunc condimentum lectus at diam tempus sodales. Nullam scelerisque auctor tellus, nec auctor ante elementum sit amet. Aenean venenatis velit eget porttitor laoreet. Aliquam aliquam libero in ante imperdiet vehicula. Donec nibh urna, tincidunt quis condimentum at, tempus sed sapien. Nunc elit dui, dignissim tincidunt mi non, ultrices ultricies erat.</p>
+
   <h2 class="title2">Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?</h2>
 +
 
 +
  <p class="texte">The only safety issue we can associate with our new Biobricks is researcher exposure to a microorganism when testing the parts. During the iGEM summer work, all safety measures were respected to avoid any problem.</p>
 +
 
 +
<h2 class="title2">Is there a local biosafety group, committee, or review board at your institution?</h2>
-
  <table class="tablecontent">
+
  <p class="texte">There is one biosafety group, leaded by a specialist in safety regulations in our institute. This person is in charge of the evaluation and implementation of the safety measures in the Instititute. Moreover, as INSA Toulouse is a five years engineer school, there are plenty of review boards that we can consult at any time to answer arising questions about biosafety. In addition, biosafety issues are regularly evaluated by the security laboratory officer. </p>
-
<tr style="background-color:#20a8da; height:50px; color:#ffffff;" >
 
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<td style="border-bottom:4px solid #e5e6e6; border-top-left-radius:9px;">TR  Title</td>
 
-
<td style="border-bottom:4px solid #e5e6e6;">TR  Title</td>
 
-
<td style="border-bottom:4px solid #e5e6e6; border-top-right-radius:9px;">TR  Title</td>
 
-
</tr>
 
-
<tr>
 
-
<td style="border-right:1px solid #e5e6e6;">Quisque</td>
 
-
<td style="border-right:1px solid #e5e6e6;">Maecenas a lorem augue</td>
 
-
<td>Maecenas a lorem augue</td>
 
-
</tr>
 
-
<tr style="border-top:1px solid #e5e6e6">
 
-
<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;">Quisque</td>
 
-
<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;">Maecenas a lorem augue</td>
 
-
<td style="border-top:1px solid #e5e6e6;">Maecenas a lorem augue</td>
 
-
</tr>
 
-
<tr>
 
-
<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;">Quisque</td>
 
-
<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;">Maecenas a lorem augue</td>
 
-
<td style="border-top:1px solid #e5e6e6;">Maecenas a lorem augue</td>
 
-
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<tr>
 
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<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;">Quisque</td>
 
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<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;">Maecenas a lorem augue</td>
 
-
<td style="border-top:1px solid #e5e6e6;">Maecenas a lorem augue</td>
 
-
</tr>
 
-
</table>
 
 +
<h2 class="title2">Do you have any other ideas how to deal with safety issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?</h2>
 +
  <p class="texte">Safety issues are usually highly specific for each project. Associating DNA parts together will always pose safety risks in case of accidental release of DNA or bacteria occur. European safety rules are pretty strict and are tailored to avoid accidental contamination of the environment or the humans manipulating these microorganisms. We think that the fact that Safety Issues are mandatory for iGEM projects is a very good way to force students (and their advisors!) to take some time evaluating (or getting help to evaluate) their specific safety (biosafety) issues.
 +
<br>When we discussed biosafety issues of our project, we analyzed the 2012 Paris-Bettencourt iGEM project named "<a href="https://2012.igem.org/Team:Paris_Bettencourt">Bware</a>", which proposes several measures to ensure that new synthetic constructions do not consitute health or environmental issues if the strain was released on the wild. Such a system would be ideal to destroy any genetic material that could be released during or after the use of <i>E. calculus</i>.
 +
</p>
</div>
</div>

Latest revision as of 21:14, 4 October 2013

logo


Safety in the Lab

Safety in INSA Toulouse

INSA Toulouse is a public engineer school in which several government research laboratories are present. Workers at INSA must follow the French rules concerning safety and environmental regulations. Most of these rules are presented in the manual “Manuel de sécurité en laboratoires” (from the WHO), which allowed us to understand and apply safety rules when manipulating microorganisms and chemicals. In addition, daily use of the Deming wheel allowed us to improve the safety rules and keep them updated.

Concerning the laboratory

Personal protective equipment

We used conventional protection all along our daily manipulating. All experiments were perfomed with gloves and a conventional lab coat, protecting the worker and avoiding contaminations. We also wear glasses when needed (UV exposition, hot water manipulations, chemicals manipulation, etc.).

Biological and Chemical Waste

After a 1h30 training for chemical waste gesture and biological hazard, we learned how to use different trash containers as described in the picture. Biological waste, potentially containing viable microorganism, were autoclaved in a special bag, then discarded. Chemical wastes are grouped by categories and eliminated via the regular procedure for all chemicals of the INSA.

Devices and Material

Here is a list of devices we used and how to use them safely:

  • Chemical Storage
    We got separated and dedicated racks for every kind of chemical product we use. As you can see on these pictures, those boxes are key-closed.
  • Ethidium bromide
    A dark room is dedicated for using EtBr and UV. This room is key-closed. Everyone entering the room must wear gloves, glasses and lab coat. Two special boxes are available to discard spoiled EtBr (one for used agarose gels, a second for gloves).
  • Biological safety cabinet
    To avoid external contamination by unwanted microorganisms on agar plates we used a Biological safety cabinet (FASTER – Ultrasafe). The cabinet is cleaned-up every morning when we arrive and every evening when we leave.
  • Electric burner
    For all sterile manipulations, we used electric burners. They are as efficient as Bunsen burners (even more), but much safer than them (limiting greatly the risks of fire hazards).
  • Chemical Hood
    To protect the user from volatile chemical compounds while dangerous manipulations, we used of two chemical hoods.
  • Water-bathes
    Use of water-bathes can be dangerous, with exposition of boiling or hot water. We used special gloves for protecting us from projections and steam. The use of water-bathes implies of course wearing glasses and lab coat. Special care was taken to switch them off every evening before leaving!
  • P2 Laboratory
    Special care have been taken to manipulate the Chromobacterium violaceum strain which is classified as a class 2 microorganism. All manipulations with this bacteria were performed in a a dedicated P2 laboratory, fully equipped for microbiology. Biological waste from C. violaceum were treated with special procedures according to the Safety Regulation Rules for Class II microorganisms.

Concerning our E. calculus project

Would E. calculus raise safety issues in terms of:

  • Researcher safety?
    In this project, we are working with biosafety level 1 microorganism (according to World Health Organization (WHO) (Laboratory Biosafety Manual) with Escherichia coli DH5-alpha, DH5-1 and XL1-Blue strains, which are considered as biologically safe for users. Some of chemical compounds are dangerous for health (EtBr, Phenolchloroform, Chloramphenicol), but as previously described, all precautions are taken to ensure security and safety for all users.

  • Public safety?
    As described in the overview, E. calculus is not designed to be used on other environments than the laboratory. Interactions with human body are not envisaged and public should also never be in contact with our modified organism. All the experiments we have performed we made according to the highest standards of safety measures. Each modification we apply to our strains should not be threatening for biodiversity and external organisms beacuse the E. coli strains that are routinely used in the laboratories are mutated in many genes, and should not be able to survive in an external environment.
    However, we are conscious that the gates we designed could potentially (and hopfully :-)) used for other biological applications, especially in metabolic engineering. In our view, these potential applications will probably be part of a process in a closed reactors, or in controlled environment, hence the safety regulations in use in the industrial context will apply. We also believe that our system present no evident potential danger, mainly because the used genes or parts were never described as potential threats to human and hopefully will never be an issue for public safety.

  • Environmental safety?
    E. calculus should not be disseminated in the environment of course. Avoiding contaminations and accidental release of the E. coli strain implies good laboratory practise, particularly for elimination of biological waste (see above).

Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?

The only safety issue we can associate with our new Biobricks is researcher exposure to a microorganism when testing the parts. During the iGEM summer work, all safety measures were respected to avoid any problem.

Is there a local biosafety group, committee, or review board at your institution?

There is one biosafety group, leaded by a specialist in safety regulations in our institute. This person is in charge of the evaluation and implementation of the safety measures in the Instititute. Moreover, as INSA Toulouse is a five years engineer school, there are plenty of review boards that we can consult at any time to answer arising questions about biosafety. In addition, biosafety issues are regularly evaluated by the security laboratory officer.

Do you have any other ideas how to deal with safety issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?

Safety issues are usually highly specific for each project. Associating DNA parts together will always pose safety risks in case of accidental release of DNA or bacteria occur. European safety rules are pretty strict and are tailored to avoid accidental contamination of the environment or the humans manipulating these microorganisms. We think that the fact that Safety Issues are mandatory for iGEM projects is a very good way to force students (and their advisors!) to take some time evaluating (or getting help to evaluate) their specific safety (biosafety) issues.
When we discussed biosafety issues of our project, we analyzed the 2012 Paris-Bettencourt iGEM project named "Bware", which proposes several measures to ensure that new synthetic constructions do not consitute health or environmental issues if the strain was released on the wild. Such a system would be ideal to destroy any genetic material that could be released during or after the use of E. calculus.