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

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                         <a href="#manPMMA" class="inner-link" data-title="Software"></a>
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                         <a href="#fluidmech" class="inner-link" data-title="Fluid Mechanics behaviour"></a>
 
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                             <h3>The lab workflow for PDMS chips</h3>
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                             <h2>The lab workflow for PDMS chips</h2>
 
                             <h4>Foto diagrama workflow</h4>
 
                             <h4>Foto diagrama workflow</h4>
 
                             <h6 class="lessmar">1-Molding of the upper half</h6>
 
                             <h6 class="lessmar">1-Molding of the upper half</h6>
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             <section id="fluidmech" class="text-center">
 
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                             <h2>Software</h2>
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                             <h2>Fluid Mechanics behaviour inside the chip</h2>
                             <p class="lead">As we have introduced in the previous section, our system is like a patchwork, with several different platforms including actuators, sensors and control elements.</p>
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                             <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 would love to experiment. This is why we created the following experiments:</p>
                            <p class="lead">Although is essential to correctly choose the programming language for the different platforms, it is mandatory to keep an eye choosing the communication protocols between all of the device’s platforms.</p>
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                            <img class= "figureimage" alt="Figure1" src="https://static.igem.org/mediawiki/2018/9/96/T--Madrid-OLM--Device--FinalPrototype--SoftwareProto.png" style="width:90%;"/>
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                            <p class="lead" style="margin-left:5%; margin-right:5%;">Figure 3: The platform’s programming languages employed and the communication protocols between all of them.</p>
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                            <p class="lead">In our circuit there are five platforms liable to be programmed:</p>
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                             <ol class="ourlist">
 
                             <ol class="ourlist">
                                 <li><p class="lead">The <b>ESP8266</b>, module in charge of all the wifi communications. We have kept the original firmware because we didn’t have time to reprogramme it during this call.</p></li>
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                                 <li><p class="lead"><b>Our mixer:</b> Inside the chip, the fluid behaves in a laminar way. There are many <a href="https://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>
                                 <li><p class="lead">In the <b>Rodeostat</b>, potentiostat responsible of the electrochemical measurements of the sensor, we modify the original firmware so it could be controlled through the Arduino Mega, instead of a computer.</p></li>
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                                 <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><p class="lead">The <b>Arduino Mega</b> controls all the device, handling the motors, receiving the Rodeostat measurements and sending them to the cloud through the WiFi module or the serial communication with the PC.</p></li>
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                                 <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>
                                 <li><p class="lead">Outside the device, the data go to <b>Firebase server</b>. The server, on one hand, gets all the data and send them to an <b>iOS design app</b>, where the final user can watch the development of the data in real time.</p></li>
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                                 <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>
                                 <li><p class="lead">Finally, a <b>PC program</b>, written in python with Qt creator, is able to communicate through the serial protocol with the device. The application let you configure 8 different motors, run protocols sequentially or inject liquid in the microfluidic chips.</p></li>
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                                 <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 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 buffer solution. We integrated the Dropsens electrode, looking forward to replicating the results obtained in the laboratory:</p></li>
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                                <a class="btn btn--primary-2 btn--sm type--uppercase" href="https://2018.igem.org/Team:Madrid-OLM/ElectrodeIntegration">
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                                        Binding the aptamers to the electrode
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                             </ol>
 
                             </ol>
                            <img class= "figureimage" alt="Figure1" src="https://static.igem.org/mediawiki/2018/6/62/T--Madrid-OLM--Device--FinalPrototype--CapturProgram.png" style="width:80%;"/>
 
                            <p class="lead">You can find the code for the PC app, the Arduino control and the Rodeostat’s modified firmware in <a href="http://github.com/OpenLabMadrid/iGEM-Madrid-OLM">our GitHub.</a> Both the iOS app and the firebase server was set up thanks to the help of <b>Marcos Hernández Cifuentes</b>.</p>
 
 
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Revision as of 10:37, 15 October 2018

Madrid-OLM

Microfluidics part of the device

Microfluidics

When the need of moving microvolumes arise 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. PDMS was our initial choice, due to its reasonable price and ease of use. As DIY and digital manufacturing constitute the basis of our hardware, we built a workflow around the PDMS chip

The lab workflow for PDMS chips

Foto diagrama workflow

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. 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.

  3. 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.

2-Molding of the lower half

The process is repeated without the negative.

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 would love to experiment. 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. 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.

  3. 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. .

  4. 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.

  5. 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 buffer solution. We integrated the Dropsens electrode, looking forward to replicating the results obtained in the laboratory:

  6. Binding the aptamers to the electrode