Team:TU Eindhoven/Safety/Our Lab

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                   <h2>Microfluidics: Introduction</h2>
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                   <h2>Our Lab</h2>
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                   <p>A substantial part of the TU Eindhoven iGEM 2014 Project is Microfluidics. Microfluidics is a technique that comprises various fields of engineering. This technique operates on a microscale and thus uses small volumes. For the encapsulation of bacterial cells – as is the case for our iGEM Project – droplet-based microfluidics is used. As extensively elaborated in the <a href="https://2014.igem.org/Team:TU_Eindhoven/Overview">General Overview Page</a>, the engineered bacteria must be brought in the proximity of PEG polymers. The ultimate goal is to verify the intended function of a cell encapsulation device. Theoretically three processes have to be accomplished:</p>
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                   <p>The laboratory of Chemical Biology (see <a href='#Fig1'>Figure 1</a> for a plan) is located in the building Helix on the TU Eindhoven campus. This laboratory is part of the department of Biomedical Engineering. The laboratory of Chemical Biology consists of two catagories of laboratories. The first one is a ML-I laboratory with biosafety level 1 (WHO BSL 1). Most of the work at the ML-I lab involves recombinant expression of proteins in <i>E. coli</i> or in yeast expression systems and the application of bacteriophages for phage display. The second one is a ML-II laboratory (cell lab) with biosafety level 2 (WHO BSL 2). Work in the ML-II lab typically involves the use of immortalized cell lines, but it can also include work with tissues or primary cells from animals or humans.</p>
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<img id='Fig1' src="https://static.igem.org/mediawiki/2014/a/ad/TU_Eindhoven_Laboratory.png" class="image_wrapper image_fr" width="1085">
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      <li>Combining a PEG solution and a bacterial culture with our engineered <i>E. coli</i> bacteria. These are the essential substances to perform the click reaction. After combining, this is called the Water Phase.</li> 
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<p style="font-size:18px;color:#CCCCCC;">Figure 1. Plan of the Laboratory of Chemical Biology.</p>
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      <li>The Water Phase has to be dispersed with an Oil Phase into droplets. This oil phase contains a fluorosurfactant in order to prevent aggregation and agglomeration of the droplets. It is crucial to form droplets with one cell in each droplet, since it will assure that each bacterial cell is encapsulated correctly – possible formation of a film is hereby avoided.</li>
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      <li>The result of the previous two processes is a droplet with PEG and engineered E. coli bacteria inside an oil phase. With a microfluidic feature (bumpy mixer) the droplet can be stirred. The droplets are collected in a chamber where the click reaction is initiated with the use of UV light.</li> 
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<p>
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<h3>Safety Equipment</h3>
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Before these processes can be performed, certain questions have to be answered and certain research has to be done. For instance, how is it possible to form droplets? What is the optimal method to recollect the content of the droplets? And numerous of other questions that need answering for instance the required flow speed of the water and oil phase and the viscosities of the phases.
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<p>The basic protection equipment that we use in the wet lab are lab coats, safety goggles and gloves. Lab coats and safety glasses are always and everywhere used in the lab (ML-I and ML-II). Gloves (mostly latex) are used when handling chemicals and for working DNase free. In the cell lab (ML-II) they are also used for working with biological materials in the biosafety cabinets. Nitrile gloves are used for handling chemicals that pass through latex or for persons allergic to latex. Cryo-gloves and heat-resistant gloves are used for handling cold and hot materials respectively. Also the fume hood is used when working with hazardous or toxic fumes.</p>
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<img id='Fig1' src="https://static.igem.org/mediawiki/2014/9/97/TU_Eindhoven_Microfluidics_Logo.jpg" class="image_wrapper image_fr" width="1085">
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<img id='Fig2' src="https://static.igem.org/mediawiki/2014/e/ed/TU_Eindhoven_Safety_Gloves.JPG" class="image_wrapper image_fr" width="1085">
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<p style="font-size:18px;color:#CCCCCC;">Figure 1. TU Eindhoven iGEM 2014 in a microfluidic device.</p>
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<img id='Fig2' src="https://static.igem.org/mediawiki/2014/d/d5/TU_Eindhoven_Safety_Rest.JPG" class="image_wrapper image_fr" width="1085">
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<p style="font-size:18px;color:#CCCCCC;">Figure 2. Safety equipment</p>
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<h4>Bibliography</h4>
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<h3>Working Equipment</h3>
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<p>Song H, Chen DL, Ismagilov RF. Reactions in droplets in microfluidic channels. Angew Chem Int Ed Engl. 2006;45:7336–7356
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<p>A lot of equipment was used during our project. The most important equipment is standard equipment like pipettes and pipettes tips to measure and transport liquids, but also the Bunsen Burner to work sterile and centrifuges to spin the samples were used. Also other equipment is used like the PCR machine for DNA amplification and gel electrophoresis apparatus for gel extraction.</p>
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Mazutis, L., Gilbert, J., Ung. W.L., Weitz, D.A., Griffiths, A.D. & Heyman J.A. (2013). Single-cell analysis and sorting using droplet-based microfluidics. Nature, 8(5), pp. 870-91.
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Song H, Chen DL, Ismagilov RF. Reactions in droplets in microfluidic channels. Angew Chem Int Ed Engl. 2006;45:7336–7356
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</p>
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<img id='Fig3' src="https://static.igem.org/mediawiki/2014/1/19/TU_Eindhoven_Equipment_1.JPG" class="image_wrapper image_fr" width="1085">
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<img id='Fig3' src="https://static.igem.org/mediawiki/2014/2/28/TU_Eindhoven_Equipment_2.JPG" class="image_wrapper image_fr" width="1085">
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<p style="font-size:18px;color:#CCCCCC;">Figure 3. Working equipment</p>
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<img id='Fig4' src="https://static.igem.org/mediawiki/2014/1/11/TU_Eindhoven_Safety1.jpg" class="image_wrapper image_fr" width="1085">
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<p style="font-size:18px;color:#CCCCCC;">Figure 4. Open top bench with amongst other pipets, pipets tips, centrifuge, Bunsen burner and PCR machine.</p>
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<img id='Fig5' src="https://static.igem.org/mediawiki/2014/0/0b/TU_Eindhoven_Safety2.jpg" class="image_wrapper image_fr" width="1085">
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<p style="font-size:18px;color:#CCCCCC;">Figure 5. Gel electrophoresis  and SDS-page place.</p>
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<img id='Fig6' src="https://static.igem.org/mediawiki/2014/e/e9/TU_Eindhoven_Safety3.jpg" class="image_wrapper image_fr" width="1085">
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<p style="font-size:18px;color:#CCCCCC;">Figure 6. Storage place for chemicals.</p>
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<img id='Fig7' src="https://static.igem.org/mediawiki/2014/b/b2/TU_Eindhoven_Safety4.png" class="image_wrapper image_fr" width="1085">
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<p style="font-size:18px;color:#CCCCCC;">Figure 7. High speed centrifuge and other lab materials.</p>
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<img id='Fig8' src="https://static.igem.org/mediawiki/2014/f/fb/TU_Eindhoven_Foto_1_onderaan_pagina_our_lab1.jpg" class="image_wrapper image_fr" width="1085">
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<p style="font-size:18px;color:#CCCCCC;">Figure 8. Working in the lab.</p>
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Latest revision as of 00:12, 18 October 2014

iGEM Team TU Eindhoven 2014

iGEM Team TU Eindhoven 2014

Our Lab

The laboratory of Chemical Biology (see Figure 1 for a plan) is located in the building Helix on the TU Eindhoven campus. This laboratory is part of the department of Biomedical Engineering. The laboratory of Chemical Biology consists of two catagories of laboratories. The first one is a ML-I laboratory with biosafety level 1 (WHO BSL 1). Most of the work at the ML-I lab involves recombinant expression of proteins in E. coli or in yeast expression systems and the application of bacteriophages for phage display. The second one is a ML-II laboratory (cell lab) with biosafety level 2 (WHO BSL 2). Work in the ML-II lab typically involves the use of immortalized cell lines, but it can also include work with tissues or primary cells from animals or humans.

Figure 1. Plan of the Laboratory of Chemical Biology.

Safety Equipment

The basic protection equipment that we use in the wet lab are lab coats, safety goggles and gloves. Lab coats and safety glasses are always and everywhere used in the lab (ML-I and ML-II). Gloves (mostly latex) are used when handling chemicals and for working DNase free. In the cell lab (ML-II) they are also used for working with biological materials in the biosafety cabinets. Nitrile gloves are used for handling chemicals that pass through latex or for persons allergic to latex. Cryo-gloves and heat-resistant gloves are used for handling cold and hot materials respectively. Also the fume hood is used when working with hazardous or toxic fumes.

Figure 2. Safety equipment

Working Equipment

A lot of equipment was used during our project. The most important equipment is standard equipment like pipettes and pipettes tips to measure and transport liquids, but also the Bunsen Burner to work sterile and centrifuges to spin the samples were used. Also other equipment is used like the PCR machine for DNA amplification and gel electrophoresis apparatus for gel extraction.

Figure 3. Working equipment

Figure 4. Open top bench with amongst other pipets, pipets tips, centrifuge, Bunsen burner and PCR machine.

Figure 5. Gel electrophoresis and SDS-page place.

Figure 6. Storage place for chemicals.

Figure 7. High speed centrifuge and other lab materials.

Figure 8. Working in the lab.

iGEM Team TU Eindhoven 2014

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