Difference between revisions of "Team:Madrid-OLM/HardawareMicrofluidics"

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     <head>
 
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         <meta charset="utf-8">
 
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         <title>Titulo del apartado</title>
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         <title>Microfluidics part of the device</title>
 
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                     </li>
 
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                     <li>
 
                     <li>
                         <a href="#story" class="inner-link" data-title="1 apartado"></a>
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                         <a href="#workflowPDMS" class="inner-link" data-title="Lab workflow for PDMS chips"></a>
 
                     </li>
 
                     </li>
 
                     <li>
 
                     <li>
                         <a href="#ceremony" class="inner-link" data-title="2 apartado"></a>
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                         <a href="#manPMMA" class="inner-link" data-title="Manufacturing the PMMA chips"></a>
 
                     </li>
 
                     </li>
 
                     <li>
 
                     <li>
                         <a href="#couple" class="inner-link" data-title="3 apartado"></a>
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                         <a href="#fluidmech" class="inner-link" data-title="Fluid Mechanics behaviour"></a>
 
                     </li>
 
                     </li>
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                     <li>
 
                     <li>
                         <a href="#reception" class="inner-link" data-title="4 apartado"></a>
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                         <a href="#plamabond" class="inner-link" data-title="Plasma Bonding"></a>
 
                     </li>
 
                     </li>
 
                     <li>
 
                     <li>
                         <a href="#rsvp" class="inner-link" data-title="5 Apartado"></a>
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                         <a href="#Injection" class="inner-link" data-title="Injection"></a>
 
                     </li>
 
                     </li>
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                      <li>
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                        <a href="#improv" class="inner-link" data-title="Further improvements"></a>
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                    </li>
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                 </ul>
 
                 </ul>
 
             </section>
 
             </section>
 
              
 
              
             <section id="home" class="cover height-100 imagebg text-center parallax" data-overlay="3">
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             <section id="home" class="tittle-secc text-center switchable feature-large">
                <div class="background-image-holder">
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                <div class="container">
                    <img alt="background" src="img/wedding-1.jpg" /> <!-- Imagen del apartado si se necesita -->
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                    <div class="row justify-content-around">
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                        <div class="col-md-8 col-lg-8">
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                            <h1 id="Teamtittle">Microfluidics</h1>
 +
                            <p class="lead">When the need of moving microvolumes arises as a mandatory requirement of design, microfluidics pops up as the one and only solution. Although there is at hand a wide range of microfluidic commercial solutions, many of them are too expensive to start experimenting with.</p>
 +
                            <p class="lead">That is why our method comes to give an alternative solution. The <a href="http://www.elveflow.com/microfluidic-tutorials/microfluidic-reviews-and-tutorials/the-poly-di-methyl-siloxane-pdms-and-microfluidics/">PDMS</a> manufacturing reveals itself as a tough rival with respect to other alternatives. Although there is at hand a wide range of microfluidic commercial solutions, many of them are too expensive to start experimenting with.</p>
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            </section>
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            <section id="workflowPDMS" class="text-center">
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                <div class="container">
 
                     <div class="row">
 
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                         <div class="col-md-10 col-lg-8 boxed boxed--border bg--secondary boxed--lg box-shadow">
                             <h1 class="h1--large">
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                             <h2>The lab workflow for PDMS chips</h2>
                                Titulo del apartado
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                            <img alt="Image1" src="https://static.igem.org/mediawiki/2018/2/24/T--Madrid-OLM--Device--FinalPrototype--Micro--workflow.png" style="width:100%;"/>
                             </h1>
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                             <h6 class="lessmar">1-Molding of the upper half</h6>
                             <p class="lead">
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                             <ol class="ourlist">
                                 Alguna descripcion si queremos
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                                <li><p class="lead"><u>Negative</u>:a laser cuts the tape that is adhered to an acetate. The remaining tape is removed carefully. The channels and the chambers, as well as the input and the output have been cutted and the negative has been created. More info about the protocols involved <a href="https://2018.igem.org/Team:Madrid-OLM/ProManufacturing#MoldM">here</a>.</p></li>
                            </p>
+
                                 <img alt="Image1" src="https://static.igem.org/mediawiki/2018/0/00/T--Madrid-OLM--Device--FinalPrototype--Micro--lasercutting.png" style="width:60%;"/>
                            <a class="btn btn--primary type--uppercase inner-link" href="#rsvp">
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                                <li><p class="lead"><u>Molding box</u>: (Find the polymerization chamber in <a href="http://github.com/OpenLabMadrid/iGEM-Madrid-OLM/tree/master/CAD/Polymerization%20chamber">our github</a>). Once the negative has been created, it is time to align the acetate with the marks in the polymerization chamber. Depending on the chosen configuration, it might be worth to place the perforated base on the bottom of the acetate.</p></li>
                                <span class="btn__text">
+
                                <li><p class="lead"><u>PDMS casting</u>: PDMS casting was made inside an lab oven most of times. Curing time depended on the drying method selected. More info about the protocols involved  <a href="https://2018.igem.org/Team:Madrid-OLM/ProManufacturing#PDMSCas">here</a>.</p></li>
                                    Posible boton a alguna zona importante
+
                                <img alt="Image1" src="https://static.igem.org/mediawiki/2018/b/b1/T--Madrid-OLM--Device--FinalPrototype--Micro--pdmsetup.png" style="width:60%;"/>
                                </span>
+
                            </ol>
                             </a>
+
                            <h6 class="lessmar">2-Molding of the lower half</h6>
 +
                            <p class="lead">The process is repeated without the negative part of the mold.</p>
 +
                            <h6 class="lessmar">3-Fixing the two halves</h6>
 +
                            <p class="lead">the selected method for fixing both halves was plasma bonding. More info about the protocols involved <a href="https://2018.igem.org/Team:Madrid-OLM/ProManufacturing#PlamsB">here</a>.</p>
 +
                            <h6 class="lessmar">4-Creating the input and the outputs</h6>
 +
                            <p class="lead">We usually used to hole punch the PDMS inlet/outlet with a needle. But we cured the PDMS with a needle inside as another negative volume for molding.More info about the protocols involved <a href="https://2018.igem.org/Team:Madrid-OLM/ProManufacturing#Closingcir">here</a>.</p>
 +
                            <h6 class="lessmar">5-Injecting fluids into the chip</h6>
 +
                             <p class="lead">Automatic controlled microvolume pressure pumps have been developed specifically for our microfluidic chips. Specific plans of the pumps design can be found in <a href="http://github.com/OpenLabMadrid/iGEM-Madrid-OLM/tree/master/CAD/Pressure%20pump">our github</a>. </p>
 
                         </div>
 
                         </div>
 
                     </div>
 
                     </div>
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             </section>
 
             </section>
 
              
 
              
             <section id="story" class="text-center">
+
             <section id="manPMMA" class="text-center">
 
                 <div class="container">
 
                 <div class="container">
 
                     <div class="row">
 
                     <div class="row">
                         <div class="col-md-10 col-lg-8">
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                         <div class="col-md-10 col-lg-8 boxed boxed--border bg--secondary boxed--lg box-shadow">
                             <h2>Apartado 1</h2>
+
                             <h2>Manufacturing the PMMA chips</h2>
                             <p class="lead">
+
                             <p class="lead">Although we are proud of having implemented an affordable workflow for developing functional PDMS chips, we manufactured PMMA chips with micromachining techniques.</p>
                                Cosas del apartado 1, se necesita meter mas divs seguramente
+
                            <p class="lead">Our University has a mechanical workshop that usually machines vacuum chambers, or metallic parts of machines, bending aluminum sheets, etc. We visited the workshop and asked the workers how to micromachine a PMMA chip with almost 0.2 mm height and 0.8 channel width. We purchased a 0.4 mm tip diameter and adapted the manufacturing to other available tools. </p>
                             </p>
+
                             <p class="lead">The input and output needed to be modified, and we used 21G needles (0.8 mm) as inlet and outlet. The fitting was made with High Performance Liquid Chromatography (HPLC) 0.8 mm tubes. They fitted tight enough to avoid leaks.</p>
 +
 
 
                         </div>
 
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             </section>
 
             </section>
 
              
 
              
             <!--MEETUPS-->
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             <section id="attendance" class="text-center">
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             <section id="fluidmech" class="text-center">
 
                 <div class="container">
 
                 <div class="container">
                     <div class="row boxed boxed--border bg--secondary boxed--lg box-shadow">
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                     <div class="row">
                         <div class="col-md-10 col-lg-10">
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                         <div class="col-md-10 col-lg-8 boxed boxed--border bg--secondary boxed--lg box-shadow">
                             <h2>Attendance to Meet Ups</h2>
+
                             <h2>Fluid Mechanics behaviour inside the chip</h2>
                             <p class="lead">The meets ups bring several advantages to the teams. It is one of the simplest ways to reach a greater number of people to whom you can explain your project. And therefore they allow you to know first-hand the projects of the rest of the teams so that you can consult and clarify doubts. These meetings also allow for more related and powerful collaborations thanks to knowing the rest of the teams in person. And last but not least, they allow you to practice exposing your project with a view to improving in the Giant Jamboree.</p>
+
                             <p class="lead">Once the workflow was designed and implemented, we focused on designing microfluidic concepts that could prove our system right. In this regard, there were some Fluid Mechanics concepts that we wanted to experiment with. This is why we created the following experiments:</p>
                            <p class="lead">Our team attended two meet ups that took place in the month of August.</p>
+
                             <ol class="ourlist">
                           
+
                                 <li><p class="lead"><b>Our mixer:</b> Inside the chip, the fluid behaves in a laminar way. There are many <a href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4634658/">papers</a> on this topic.We wanted to test this experimentally. And that is why we created a mixer. We could study how the fluid behaves in the conditions of a mixer. Our mixer is just an example on how microfluidic components can be small enough to be modularly assembled in series or in parallel as an electronic component.</p></li>
                             <div id="meetsups" class="tabs-container" data-content-align="center">
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                                <img alt="Image1" src="https://static.igem.org/mediawiki/2018/0/05/T--Madrid-OLM--Device--FinalPrototype--Micro--circuit1.png" style="width:35%;"/>
                                 <ul class="tabs">
+
                                <li><p class="lead"><b>Flow separation tests:</b> We have designed four experiments to study the behaviour of our flow under different circumstances. The flow circulates towards a triangle, a circle, a throat and the shape of a heart. This will show us how the flow behaves under certain circumstances. Its immediate consequences affect the design of chambers or any microchannel widening.</p></li>
                                    <li class="active">
+
                                <img alt="Image1" src="https://static.igem.org/mediawiki/2018/6/64/T--Madrid-OLM--Device--FinalPrototype--Micro--circuit2.png" style="width:35%;"/>
                                        <div class="tab__title">
+
                                <li><p class="lead"><b>Droplet generation tests:</b> Generating droplets is one of the milestones of microfluidics. Droplets are small volumes of sample moving as small drops in an arranged and harmonic way. It is much more than just beautiful. The main task of this chip is to study how a fluid and air pressure gradients can work together in the same room. The design pushes to the limit the available capabilities of our device.</p></li>
                                            <span class="h5">Marseille</span>
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                                <img alt="Image1" src="https://static.igem.org/mediawiki/2018/7/7f/T--Madrid-OLM--Device--FinalPrototype--Micro--circuit3.png" style="width:35%;"/>
                                        </div>
+
                                <li><p class="lead"><b>Tree and mixer test:</b> We have designed a large PMMA chip to work as a sample on how fluid behaves when flow is separated into different branches of a tree. The aim of this experiment is to study the laminar flow, and how it behaves when it arrived to the central chamber. On the exact opposite side, a negative relative pressure will be generated to study how it behaves in an alternative “negative relative pressure” tree. In this experiment there are two sides of a chip. Both of them are experimentally equivalent.  </p></li>
                                        <div class="tab__content">
+
                                <img alt="Image1" src="https://static.igem.org/mediawiki/2018/7/7f/T--Madrid-OLM--Device--FinalPrototype--Micro--circuit4.png" style="width:35%;"/>
                                            <h3>Mediterranean Meet Up (Marseille)</h3>
+
                                <li><p class="lead"><b>A chip adapted to Dropsens GNP110 electrode:</b> We manufactured via regular CNC milling, adapted to micromachining, the housing for a Dropsens GNP110 electrode. A <a href="http://www.sciencedirect.com/science/article/pii/S0956566317304013?via%3Dihub">paper</a> proved our arrangement to be functional. We manufactured a two part chip. The upper side was micromilled with a 0.4 mm tool, with a custom made circuit for injecting the protein solution, ferricyanide and a buffer solution.We integrated the Dropsens electrode, looking forward to replicating the results obtained in the laboratory:</p></li>
                                            <img alt="Image1" src="https://static.igem.org/mediawiki/2018/9/9a/T--Madrid-OLM--Collaboration--Marseille--All.png" style="width:80%;"/>
+
                               
                                            <p class="lead">This meet up took place between Thursday 9 and Friday 10 August in the city of Marseille and was organized by the team of this city, the Aix-Marseille. In it, all the teams from the Mediterranean area were summoned, so they ended up attending both France and Spain. Francisco, Gonzalo and Ivan attended this meet up on behalf of the team.</p>
+
                                <img alt="Image1" src="https://static.igem.org/mediawiki/2018/8/8f/T--Madrid-OLM--Device--FinalPrototype--Micro--circuit5.png" style="width:55%;"/>
                                            <p class="lead">The event began with a reception in which all teams had their first informal contact. Subsequently, a 5 minutes presentation of each of the projects was organized, summarizing the concept on which the proyect was based, the work carried out and the plans to complete it. After the exhibition, each team answered the questions that could have the jury and the rest of the teams.</p>
+
                                <br/>
                                            <img alt="Image2" src="https://static.igem.org/mediawiki/2018/4/49/T--Madrid-OLM--Collaboration--Marseille--Exp.png" style="width:70%;"/>
+
                                <a class="btn btn--primary-2 btn--sm type--uppercase" href="https://2018.igem.org/Team:Madrid-OLM/ElectrodeIntegration">
                                            <p class="lead">Later they invited us to a joint meal. In the afternoon it was organized a sightseeing tour of the city, but due to inclement weather it had to be suspended, although we could enjoy a pleasant evening of board games and conversations with the rest of the teams. At night a meeting was also organized in a pub to end a day where we had met many new people.</p>
+
                                    <span class="btn__text">
                                            <p class="lead">The second (and last day) had two interesting activities. The first was the meetings with different experts from the university, which privately asked each team about controversial issues. To see if these issues had been taken into account and the possible solutions and consequences were debated.</p>
+
                                        Binding the aptamers to the electrode
                                            <img alt="Image3" src="https://static.igem.org/mediawiki/2018/2/22/T--Madrid-OLM--Collaboration--Marseille--Experts.png" style="width:70%;"/>
+
                                    </span>
                                            <p class="lead">To close the event, a poster presentation session was held, which was very enriching to know how to present our project in Boston..</p>
+
                                </a>
                                            <p class="lead">One of the things that were achieved in this meet up was the agreement with the UPF-CRG-Barcelona group to organize an activity in their meet up. Activity that is explained in the following session (and that gave rise to a greater collaboration).</p>
+
                               
                                            <img alt="Image4" src="https://static.igem.org/mediawiki/2018/2/2b/T--Madrid-OLM--Collaboration--Marseille--Poster.png" style="width:70%;"/>
+
                            </ol>
                                            <br/>
+
                        </div>
                                        </div>
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                    </div>
                                    </li>
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                                    <li>
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                </div>
                                        <div class="tab__title">
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                <!--end of container-->
                                            <span class="h5">Barcelona</span>
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            </section>
                                        </div>
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                                        <div class="tab__content">
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            <section id="plamabond" class="text-center">
                                            <h3>Spanish Meet Up (Barcelona)</h3>
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                <div class="container">
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                    <div class="row">
                                            <p class="lead">This meet up took place between Saturday 18 and Monday 20 August in the city of Barcelona and was organized by one of the two teams of this same city, the UPF-CRG-Barcelona. In it, all the teams from Spain were called, which all but the Valencia team attended. To this meet up they were representing the whole team: Laura, Iván and Francisco. The meet up took place in the Biomedical Research Park of Barcelona (PRBB), center where the host team operates and where they started with a welcome activity. This activity consisted of a game where each one of the assistants of each team presented themselves, so that the subsequent environment was much more relaxed.</p>
+
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                                            <p class="lead">The next activity was the one organized by our team. In this activity we proposed to carry out a collaborative guide in which each group would give the main ideas on how they have solved the main problems that have arisen in iGEM. The structure we proposed was followed. We collected these ideas and wrote a complete guide. We explain this collaboration in a later section of the collaborations. After this, some playful activities were carried out on the beach, including volleyball games. </p>
+
                            <h2>Plasma bonding</h2>
                                            <img alt="Image6" src="https://static.igem.org/mediawiki/2018/5/5e/T--Madrid-OLM--Collaboration--BCN--Voley.png" style="width:65%;"/>
+
                            <img alt="Image1" src="https://static.igem.org/mediawiki/2018/e/e3/T--Madrid-OLM--Device--FinalPrototype--Micro--plasma.gif" style="width:75%;"/>
                                            <p class="lead">During the morning of the second day, a more detailed presentation of each team's project was made. There was 30 minutes to expose each project in detail. This activity was very useful because the rest of the teams, besides getting to know the details of each project well, could intervene to give advice on how to make the presentations. Each team took a series of points in which they could be reinforced in the face of the presentation of the Giant Jamboree.</p>
+
                            <p class="lead">After setting the microwave up for treating the chips with plasma, we got some results that might serve as an illustration of the process. Other documentation can be found <a href="http://arxiv.org/ftp/arxiv/papers/1807/1807.06784.pdf">here</a>. As we explain in the protocols section, we used a 700W microwave, modded to fit our requirements, as we explain in the protocols section:</p>
                                            <p class="lead">In the afternoon the time was taken to take a tour in the tourist center of the city and ended up having dinner with a picnic in the Parc Guell. A viewpoint that have views of the entire city at night.</p>
+
                             <a class="btn btn--primary-2 btn--sm type--uppercase" href="https://2018.igem.org/Team:Madrid-OLM/ProManufacturing#PlamsBn">
                                            <img alt="Image7" src="https://static.igem.org/mediawiki/2018/7/78/T--Madrid-OLM--Collaboration--BCN--Tour.png" style="width:60%;"/>
+
                                            <br/>
+
                                            <p class="lead">During the morning of the last day, to close the meet up, we were able to attend to some enriching expert talks. They offered us their views on synthetic biology and they explained the works in which they are immersed today.</p>
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                                            <img alt="Image8" src="https://static.igem.org/mediawiki/2018/e/e3/T--Madrid-OLM--Collaboration--BCN--Bye.png" style="width:70%;"/>
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                             <a class="btn btn--primary-2 type--uppercase inner-link" href="#meetsups">
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                                 <span class="btn__text">
 
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                                     Back to Meet up Index
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                                     Plasma Bonding Protocol
 
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                            <br/><br/>
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                            <p class="lead">We finally configured the microwave to half of its power approximately, inserted a 100ml glass of water and 20 seconds of treatment. After these parameters were established, we got the following results.</p>
 +
                            <p class="lead">One of the indicators that show that plasma is treating the PDMS correctly is the modification of the surface tension of the water on a PDMS surface.</p>
 +
                            <img alt="Image1" src="https://static.igem.org/mediawiki/2018/f/f5/T--Madrid-OLM--Device--FinalPrototype--Micro--surfacetension.png" style="width:70%;"/>
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                            <h2>Injection</h2>
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                            <img alt="Image1" src="https://static.igem.org/mediawiki/2018/2/23/T--Madrid-OLM--Device--FinalPrototype--Micro--pumpswork.png" style="width:75%;"/>
 +
                            <p class="lead">One of the improvements of the second prototype with respect to the initial is centered in the pressure system. It has the capability of displacing liquid volumes in the order of microliters. Our pressure pump has an unique arrangement, and it has been designed to be affordable and precise enough to govern the physical parameters involved in microfluidics mechanics.  </p> 
 +
                            <p class="lead">Further information can be found in <a href="http://github.com/OpenLabMadrid/iGEM-Madrid-OLM/tree/master/CAD/Pressure%20pump">our Github</a>. </p>
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                             <h2>Organization of a Meet Up: Another European Meet Up</h2>
+
                             <h2>Further improvements</h2>
                            <h5>Reason</h5>
+
                             <p class="lead">Although the microfluidic chip is quite similar to what we consider a final version, there are many situations that we want to warn about to anyone who wants to replicate our setup.</p>
                             <p class="lead">After attending two meet ups we realized that these meetings are very enriching experiences in preparation for the Giant Jamboree. One of the main reasons for these meet ups is to get to know and know the project of other teams. Other reason is that its also serves to practice the presentations of the project and its results. And finally serve to open the network of contacts beyond the contest itself.</p>
+
                             <p class="lead">Microfluidics does not always behave as we expect. DIY manufacturing is close to artisanry. Getting to a point in which replicability is expected is hard. It requires a lot of time and effort to master the technique. </p>
                            <p class="lead">We realized that the events to which more teams come are usually the first ones to be organized, and in these although if they serve to obtain collaborations, do not fulfill so much the function of practicing to present the results of your project due to at the beginning of the summer there are no results.</p>
+
                             <p class="lead">PDMS has a very positive side for DIY manufacturers: it is affordable and resilient. It is easy to understand and a good way of learning microfluidics.</p>
                             <p class="lead">This year the European meet up was on July 20. Date in which novice teams, without sufficient contacts do not yet know how to deal with these events and do not have the necessary funds to attend. And teams with more experience do not yet have conclusive results. With this context we proposed this ambitious project: Make another European meet up. We invited all the teams of the continent in mid-September. A date that was supposed to be the best for the organization of new teams and with the possibility of simulating the final event.</p>
+
                             <p class="lead">On the other hand, PMMA micromachining and precision manufacturing involve higher costs and a dependence to a mechanical workshop. You will not implement designs as fast as you can with the workflow that we have developed for PDMS, with the laser, the plasma bonding and the polymerization chamber.</p>
                           
+
                             <p class="lead">We would love to share a project for anyone to replicate a microfluidics chip in the most affordable and optimal way. DIY tools are capricious and sometimes they do not behave as we expect them to do. </p>
                            <h5>Event</h5>
+
                             <p class="lead">By repairing and refining DIY tools, we have learn a lot of machine design, manufacturing and biodevices design. We consider that DIY is the best way of learning anything. This is the reason why we would love to share our spirit and encourage any interested person to overcome these difficulties and experience the satisfaction of designing, manufacturing and searching beyond the immediate reality.</p>
                            <p class="lead">The event was organized between September 14 and 16. Although this event was given as much publicity as possible and contact was made with all possible teams in Europe, attendance was low due to several factors:</p>
+
                            <ul>
+
                                <li>
+
                                    <p class="lead"><b>- Spain is geographically at one end of Europe and travel is expensive.</b></p>     
+
                                </li>
+
                                <li>
+
                                    <p class="lead"><b>- The teams were short of funds and were saving for the trip to Boston.</b></p>
+
                                </li>
+
                                <li>
+
                                    <p class="lead"><b>- The event was announced at the beginning of September, many teams did not have room to maneuver in their organization to attend.</b></p>     
+
                                </li>
+
                            </ul>
+
                            <br/>
+
                             <p class="lead">Despite these inconveniences we had assistance from several participants of some teams (all of them came from within the Spanish territory thanks to the facilities for transport). Although there was an extensive program of activities, these had to be adjusted and reorganized so that the maximum number of attendees could enjoy them.</p>
+
                       
+
                            <h5>Activities</h5>  
+
                             <p class="lead">The event began on Friday the 14th in the afternoon with the main course. In it, after the reception of the attendees we gave way to a series of very interesting expert presentations. We were able to count on the attendance of the following personalities</p>
+
                            <p class="lead"><u><b>Victor de Lorenzo</b></u></p>
+
                            <p class="lead">He is one of the Spanish most relevant researchers in the Synthetic Biology field. He work in the National Biotechnology Center (CNB) a part of the Superior Council of Scientific Investigations (CSIC). His laboratory is specialized in solving environmental issues using genetically engineered Pseudomonas putida. He gave us an interesting talk about <i>"Synthetic Biology in the rescue of the planet Earth"</i>.</p>
+
                            <img alt="Image5" src="https://static.igem.org/mediawiki/2018/e/ee/T--Madrid-OLM--Collaboration--Madrid--Conference1.png" style="width:70%;"/>
+
                           
+
                            <p class="lead"><u><b>Saul Ares</b></u></p>
+
                            <p class="lead">He is a researcher of the CNB too. His work is central in the mathematical modeling of Synthetic Biology systems. He has also collaborated with previous iGEM teams. His presentation talked about <i>"How do cyanobacteria count to ten"</i>.</p>
+
                            <img alt="Image5" src="https://static.igem.org/mediawiki/2018/7/7e/T--Madrid-OLM--Collaboration--Madrid--Conference2.png" style="width:70%;"/>
+
                           
+
                            <p class="lead"><u><b>Krzysztof Wabnik</b></u></p>
+
                            <p class="lead">He is a Young Investigator Researcher of the Center for Biotechnology and Plant Genomics (CBGP) at the Polytechnic University of Madrid (UPM). The group he is leading uses multilevel computer model simulations, synthetic biology experiments and microfluidics. The name of his presentation was: <i>"Combining computational and synthetic biology approaches to understand the dynamics of plant hormone signaling circuits"</i>.</p>
+
                            <img alt="Image5" src="https://static.igem.org/mediawiki/2018/2/2d/T--Madrid-OLM--Collaboration--Madrid--Conference3.png" style="width:70%;"/>
+
                           
+
                             <p class="lead">Despite these inconveniences we had assistance from several participants of some teams (all of them came from within the Spanish territory thanks to the facilities for transport). Although there was an extensive program of activities, these had to be adjusted and reorganized so that the maximum number of attendees could enjoy them.</p>
+
                            <img alt="Image5" src="https://static.igem.org/mediawiki/2018/4/4a/T--Madrid-OLM--Collaboration--Madrid--Conference4.png" style="width:70%;"/>
+
                            <p class="lead">After the talks a social event was held in which the attending teams could relax having dinner in the center of Madrid with one of the most typical activities here: go out for tapas.</p>
+
                            <p class="lead">The second day was inaugurated with a visit to the university to show the facilities where our team works and it was explained which procedures are used in each of the sections that were visited.</p>
+
                             <p class="lead">Subsequently, the presentations of the projects were made. A simulation of the presentation was carried out as it should be done in Boston, to learn how to take the time and try to correct the excesses or lack of time in each section that was exposed.</p>
+
                            <p class="lead">In the afternoon there was a tour around Madrid, where the most iconic places of the city were shown. And although there were activities scheduled for the next day, this was the activity that closed the meet up, since the attending teams had organized to return to their cities after this tour.</p>
+
                            <img alt="Image5" src="https://static.igem.org/mediawiki/2018/3/30/T--Madrid-OLM--Collaboration--Madrid--Tour.png" style="width:70%;"/>
+
                            <h5>Conclusion</h5>
+
                            <p class="lead">It was an event that few people could attend, due to the conditions explained above, but from which the attendees took much advantage. Holding such an event in September could be much more crowded if it was announced from the beginning of the summer and allowed the rest of the teams to organize around it. As it provides a series of very useful tools for the presentation of Boston.</p>
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Latest revision as of 01:47, 18 October 2018

Madrid-OLM

Microfluidics part of the device

Microfluidics

When the need of moving microvolumes arises as a mandatory requirement of design, microfluidics pops up as the one and only solution. Although there is at hand a wide range of microfluidic commercial solutions, many of them are too expensive to start experimenting with.

That is why our method comes to give an alternative solution. The PDMS manufacturing reveals itself as a tough rival with respect to other alternatives. Although there is at hand a wide range of microfluidic commercial solutions, many of them are too expensive to start experimenting with.

The lab workflow for PDMS chips

Image1
1-Molding of the upper half
  1. Negative:a laser cuts the tape that is adhered to an acetate. The remaining tape is removed carefully. The channels and the chambers, as well as the input and the output have been cutted and the negative has been created. More info about the protocols involved here.

  2. Image1
  3. Molding box: (Find the polymerization chamber in our github). Once the negative has been created, it is time to align the acetate with the marks in the polymerization chamber. Depending on the chosen configuration, it might be worth to place the perforated base on the bottom of the acetate.

  4. PDMS casting: PDMS casting was made inside an lab oven most of times. Curing time depended on the drying method selected. More info about the protocols involved here.

  5. Image1
2-Molding of the lower half

The process is repeated without the negative part of the mold.

3-Fixing the two halves

the selected method for fixing both halves was plasma bonding. More info about the protocols involved here.

4-Creating the input and the outputs

We usually used to hole punch the PDMS inlet/outlet with a needle. But we cured the PDMS with a needle inside as another negative volume for molding.More info about the protocols involved here.

5-Injecting fluids into the chip

Automatic controlled microvolume pressure pumps have been developed specifically for our microfluidic chips. Specific plans of the pumps design can be found in our github.

Manufacturing the PMMA chips

Although we are proud of having implemented an affordable workflow for developing functional PDMS chips, we manufactured PMMA chips with micromachining techniques.

Our University has a mechanical workshop that usually machines vacuum chambers, or metallic parts of machines, bending aluminum sheets, etc. We visited the workshop and asked the workers how to micromachine a PMMA chip with almost 0.2 mm height and 0.8 channel width. We purchased a 0.4 mm tip diameter and adapted the manufacturing to other available tools.

The input and output needed to be modified, and we used 21G needles (0.8 mm) as inlet and outlet. The fitting was made with High Performance Liquid Chromatography (HPLC) 0.8 mm tubes. They fitted tight enough to avoid leaks.

Fluid Mechanics behaviour inside the chip

Once the workflow was designed and implemented, we focused on designing microfluidic concepts that could prove our system right. In this regard, there were some Fluid Mechanics concepts that we wanted to experiment with. This is why we created the following experiments:

  1. Our mixer: Inside the chip, the fluid behaves in a laminar way. There are many papers on this topic.We wanted to test this experimentally. And that is why we created a mixer. We could study how the fluid behaves in the conditions of a mixer. Our mixer is just an example on how microfluidic components can be small enough to be modularly assembled in series or in parallel as an electronic component.

  2. Image1
  3. Flow separation tests: We have designed four experiments to study the behaviour of our flow under different circumstances. The flow circulates towards a triangle, a circle, a throat and the shape of a heart. This will show us how the flow behaves under certain circumstances. Its immediate consequences affect the design of chambers or any microchannel widening.

  4. Image1
  5. Droplet generation tests: Generating droplets is one of the milestones of microfluidics. Droplets are small volumes of sample moving as small drops in an arranged and harmonic way. It is much more than just beautiful. The main task of this chip is to study how a fluid and air pressure gradients can work together in the same room. The design pushes to the limit the available capabilities of our device.

  6. Image1
  7. Tree and mixer test: We have designed a large PMMA chip to work as a sample on how fluid behaves when flow is separated into different branches of a tree. The aim of this experiment is to study the laminar flow, and how it behaves when it arrived to the central chamber. On the exact opposite side, a negative relative pressure will be generated to study how it behaves in an alternative “negative relative pressure” tree. In this experiment there are two sides of a chip. Both of them are experimentally equivalent.

  8. Image1
  9. A chip adapted to Dropsens GNP110 electrode: We manufactured via regular CNC milling, adapted to micromachining, the housing for a Dropsens GNP110 electrode. A paper proved our arrangement to be functional. We manufactured a two part chip. The upper side was micromilled with a 0.4 mm tool, with a custom made circuit for injecting the protein solution, ferricyanide and a buffer solution.We integrated the Dropsens electrode, looking forward to replicating the results obtained in the laboratory:

  10. Image1
    Binding the aptamers to the electrode

Plasma bonding

Image1

After setting the microwave up for treating the chips with plasma, we got some results that might serve as an illustration of the process. Other documentation can be found here. As we explain in the protocols section, we used a 700W microwave, modded to fit our requirements, as we explain in the protocols section:

Plasma Bonding Protocol

We finally configured the microwave to half of its power approximately, inserted a 100ml glass of water and 20 seconds of treatment. After these parameters were established, we got the following results.

One of the indicators that show that plasma is treating the PDMS correctly is the modification of the surface tension of the water on a PDMS surface.

Image1

Injection

Image1

One of the improvements of the second prototype with respect to the initial is centered in the pressure system. It has the capability of displacing liquid volumes in the order of microliters. Our pressure pump has an unique arrangement, and it has been designed to be affordable and precise enough to govern the physical parameters involved in microfluidics mechanics.

Further information can be found in our Github.

Further improvements

Although the microfluidic chip is quite similar to what we consider a final version, there are many situations that we want to warn about to anyone who wants to replicate our setup.

Microfluidics does not always behave as we expect. DIY manufacturing is close to artisanry. Getting to a point in which replicability is expected is hard. It requires a lot of time and effort to master the technique.

PDMS has a very positive side for DIY manufacturers: it is affordable and resilient. It is easy to understand and a good way of learning microfluidics.

On the other hand, PMMA micromachining and precision manufacturing involve higher costs and a dependence to a mechanical workshop. You will not implement designs as fast as you can with the workflow that we have developed for PDMS, with the laser, the plasma bonding and the polymerization chamber.

We would love to share a project for anyone to replicate a microfluidics chip in the most affordable and optimal way. DIY tools are capricious and sometimes they do not behave as we expect them to do.

By repairing and refining DIY tools, we have learn a lot of machine design, manufacturing and biodevices design. We consider that DIY is the best way of learning anything. This is the reason why we would love to share our spirit and encourage any interested person to overcome these difficulties and experience the satisfaction of designing, manufacturing and searching beyond the immediate reality.