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="title3">Worker protections</h3>
+
<h3 class="title3left530">Personal protective equipment</h3>
-
   <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>
+
   <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>
-
  <h3 class="title3">Waste gesture</h3>
+
<img src="https://static.igem.org/mediawiki/2013/2/2b/Hang.png" class="imgcontentright" />
 +
<div class="clear"></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.  
+
 
-
Biological wastes, potentially containing viable microorganism, are firstable autoclaved in a special bag, then discard on the classical way.
+
 
-
Chemical waste are regrouped and treated every month together with wastes of other labs of the campus.</p>
+
<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>
   <h3 class="title3">Devices and Material</h3>
-
   <p class="texte">In this part, we’ll list all devices we use and how to use them safely :</p>
+
   <p class="texte">Here is a list of devices we used and how to use them safely:</p>
 +
 
 +
 
 +
 
    
    
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     <ul class="circlearrow">
     <ul class="circlearrow">
       <li> <span class="title3">Chemical Storage</span></br>
       <li> <span class="title3">Chemical Storage</span></br>
-
<span  class="texte">We got separated and dedicated racks for every kind of chemical product we use. As you can see on pictures, those boxes are key-closed and gesture of bottles and product is made when we used them.</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">Ethydium bromure room</span></br>
+
  </div>
-
<span class="texte">A room is dedicated for using EtBr, while this reactant has been proved as highly mutagenic and dangerous for user. This room is key-closed and gloves, glasses and lab coat are compulsory to enter in. Two special boxes, for gesture of spoiled EtBr wastes, are putting at disposition of users for future and special treatments.</span></li>
+
 
-
<br>
+
<img src="https://static.igem.org/mediawiki/2013/7/77/Inflammable_-_75px.png" class="imgcontentleft" />
 +
<img src="https://static.igem.org/mediawiki/2013/c/cb/Acides_-_75px.png" class="imgcontentleft" />
 +
<img src="https://static.igem.org/mediawiki/2013/0/06/Bases_-_75px.png" class="imgcontentleft" />
 +
 
 +
<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>
       <li> <span class="title3">Biological safety cabinet</span></br>
-
<span class="texte">To avoid external contamination and limiting exposure to biological material, we use a Biological safety cabinet (FASTER – Ultrasafe) which is controlled every year. It works according to the principle of laminar flux circulation and filter, in such a way microorganism are stuck into the cabinet, and external contaminations are impossible. This cabinet is cleaned-up every morning when we arrive and every evening when we leave.</span></li>
+
<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>
-
<br>
+
 
 +
</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>
       <li> <span class="title3">Electric burner</span></br>
-
<span class="texte">For all manipulations, we also use electric burners, which are as efficient as Bunsen burners (even more), but much more safe than them (this way we limit fire hazards)</span></li>
+
<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>
-
<br>
+
</ul>
-
       <li> <span class="title3">Fume Cupboards</span></br>
+
 
-
<span class="texte">To protect the user from volatile chemical compounds while dangerous manipulations, we dispose of two Fume cupboards.</span></li>
+
  </div>
-
<br>
+
 
 +
<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>
       <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 also use special gloves protecting us from projections and steam. The use of water-bathes implies of course wearing glasses and lab coat.</span></li>
+
<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>
 +
 
 +
  </div>
 +
 
 +
<img src="https://static.igem.org/mediawiki/2013/5/5c/Hot_bathe.png" class="imgcontentleft" />
 +
 
 +
<div class="clear"></div>
 +
 
 +
<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>
     </ul>
     </ul>
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   </div>
   </div>
 +
<img src="https://static.igem.org/mediawiki/2013/1/19/P2_lab.png" class="imgcontentleft" />
<div class="clear"></div>
<div class="clear"></div>
-
   <h2 class="title2">Concerning <i>E. calculus</i> and project</h2>
+
   <h2 class="title2">Concerning our <i>E. calculus</i> project</h2>
-
   <h2 class="title2">Would <i>E. calculus</i> raise safety issues in terms of :</h2>
+
   <h2 class="title2">Would <i>E. calculus</i> raise safety issues in terms of:</h2>
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     <ul class="circlearrow">
     <ul class="circlearrow">
       <li> <span class="title3">Researcher safety?</span></br>
       <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"> Laboratory Biosafety Manual</a>) with Escherichia coli DH5α and DH51 strains, which are considered as biologically safe for users. Some of chemical compounds used could be dangerous for health (EtBr, Phenolchloroform..), but as previously described, all precautions are taken to ensure security and safety for all users.</span></li>
+
<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>
<br>
       <li> <span class="title3">Public safety?</span></br>
       <li> <span class="title3">Public safety?</span></br>
-
<span class="texte">As described in the overview, <i>E. calculus</i> should not go out of the lab. Interactions with human body are not envisaged and public should also never be in contact with our modified organism. Every manipulations we proceed are made under great control, in an enclose environment. Each modification we apply to our strains should not be threatening for biodiversity and external organisms. Furthermore, our strains are engineered and should not be able to survive in an external environment.
+
<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>
-
<br>If a direct application is find for <i>E. calculus</i> (or just for logic gates we create and test), it will be generally a part of a process in closed reactors, or in controlled environment. We also believe that our system is really safe and could never be an issue for public safety.</span></li>
+
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>
<br>
       <li> <span class="title3">Environmental safety?</span></br>
       <li> <span class="title3">Environmental safety?</span></br>
-
<span class="texte"><i>E. calculus</i> should also not be deployed in environment and will stay in the lab, at least for instance. Of course, avoiding contaminations and spreading implies a good gesture of biological waste (see the part above “waste gesture”). Furthermore, use of special strain permit us to be sure that none of our modified organisms are spread out in the wild.</span></li>
+
<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>
     </ul>
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   <h2 class="title2">Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?</h2>
   <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 microorganism when testing the parts, but as wrote earlier, all precautions are respected to avoid every problem. Our parts are also tested in different contexts, to assure their full functionality and safety whenever another team wants to use them.</p>
+
   <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>
  <h2 class="title2">Is there a local biosafety group, committee, or review board at your institution?</h2>
-
   <p class="texte">Our institute disposes of one person who is relevant for security and safety in the lab, but we do not have any group or committee concerning biosafety in general. However, we possess review boards we can consult any time to answer our question about biosafety. In addition, biosafety was evaluated by laboratory responsibles who do not have any restriction letting us work in the lab on this project.</p>
+
   <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>
  <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>
  <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">Our team does not have any local biosafety group or committee, information and documentation about biosafety and condition of application were pretty hard to find. However, we have the luck to work in a laboratory which already got all equipments and protocols to ensure a great Biosafety. It could be a good improvement for future iGEM competitions if iGEM staff could provide us way to ensure that safety is good enough to take part to the competition, like a “safety check-list”, or by putting some “limitations” about the manipulations, telling us what is dangerous or forbidden. (forbidden strains, forbidden modifications, dangerous and “key” manipulations…).
+
   <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>We also focused on the 2012 Paris-Bettencourt iGEM project, named “<a href="https://2012.igem.org/Team:Paris_Bettencourt">Bware</a>”, which proposes multiple ways to ensure that new synthetic constructions could never be an issue for health or environment, even if the strain if unfortunately released on the wild. For us, work of team Bettencourt is really important and should potentially be integrated on future iGEM standards and rules.
+
<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>
</p>

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.

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