Difference between revisions of "Team:Mingdao/Applied Design"

 
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    <a class="w3-bar-item w3-button w3-hide-medium w3-hide-large w3-right w3-padding-large w3-hover-white w3-large w3-red" href="javascript:void(0);" onclick="myFunction()" title="Toggle Navigation Menu"><i class="fa fa-bars"></i></a>
 
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    <a href="https://2017.igem.org/Team:Mingdao" class="w3-bar-item w3-button w3-padding-large w3-white">Home</a>
 
    <a href="https://2017.igem.org/Team:Mingdao" class="w3-bar-item w3-button w3-padding-large w3-white">Home</a>
    <a href="#" class="w3-bar-item w3-button w3-hide-small w3-padding-large w3-blue w3-hover-white" style="text-decoration: none; color: white">Achievements</a>
+
    <a href="https://2017.igem.org/Team:Mingdao/Achievements" class="w3-bar-item w3-button w3-hide-small w3-padding-large w3-blue w3-hover-white" style="text-decoration: none; color: white">Achievements</a>
 
    <a href="https://2017.igem.org/Team:Mingdao/Demonstrate" class="w3-bar-item w3-button w3-hide-small w3-padding-large w3-hover-white" style="text-decoration: none; color: white">Project</a>
 
    <a href="https://2017.igem.org/Team:Mingdao/Demonstrate" class="w3-bar-item w3-button w3-hide-small w3-padding-large w3-hover-white" style="text-decoration: none; color: white">Project</a>
 
    <a href="https://2017.igem.org/Team:Mingdao/HP/Silver" class="w3-bar-item w3-button w3-hide-small w3-padding-large w3-hover-white" style="text-decoration: none; color: white">Human-Practice</a>
 
    <a href="https://2017.igem.org/Team:Mingdao/HP/Silver" class="w3-bar-item w3-button w3-hide-small w3-padding-large w3-hover-white" style="text-decoration: none; color: white">Human-Practice</a>
    <a href="#" class="w3-bar-item w3-button w3-hide-small w3-padding-large w3-hover-white" style="text-decoration: none; color: white">Notebook</a>
+
    <a href="https://2017.igem.org/Team:Mingdao/Notebook" class="w3-bar-item w3-button w3-hide-small w3-padding-large w3-hover-white" style="text-decoration: none; color: white">Notebook</a>
 
    <a href="https://2017.igem.org/Team:Mingdao/Attributions" class="w3-bar-item w3-button w3-hide-small w3-padding-large w3-hover-white" style="text-decoration: none; color: white;">Attributions</a>
 
    <a href="https://2017.igem.org/Team:Mingdao/Attributions" class="w3-bar-item w3-button w3-hide-small w3-padding-large w3-hover-white" style="text-decoration: none; color: white;">Attributions</a>
 
  </div>
 
  </div>
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                     <br><br>
 
                     <br><br>
 
                     <h1 class="display-3">Prototype</h1>
 
                     <h1 class="display-3">Prototype</h1>
                     <h4 class="lead">Not only in the lab,<br> we brought our design to reality</h4>
+
                     <h4 class="lead">Not only in the lab,<br> we brought our design into reality</h4>
 
                     <hr class="my-4">
 
                     <hr class="my-4">
 
                 </div>
 
                 </div>
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             <div class="row">
 
             <div class="row">
 
                 <div class="col-md-10 col-sm-12" style="display: block; margin: auto">
 
                 <div class="col-md-10 col-sm-12" style="display: block; margin: auto">
 +
<div class="card" style="margin-bottom: 3rem">
 +
  <div class="card-header text-center" style="color: white; letter-spacing: 0.3rem;font-size:1.8rem;background-color:#1dd667">
 +
Acid-resistant Microparticles
 +
  </div>
 +
  <div class="card-body">
 +
  <!-- PART start -->
  
 +
  <h4 class="w3-padding-24">
 +
  To make probiotics as a good food supplement, one may consider the engineered microbes need to pass through gastric acid in the digestive system. Calcium alginate nanoparticles are good carriers for delivery which are acid-resistant (pH=2) and releasing in basic environment (pH=9). The following is our protocol in the microencapsulation process.
 +
  </h4>
 +
<h3><b>Microencapsulation</b></h3>
 +
<h4 class="w3-padding-24">
 +
  <ol>
 +
  <li>Make 0.25% alginate: 0.25 g of Alginic acid sodium salt (SIGMA-ALDRICH) in 100 ml of H<sub>2</sub>O. Heat at 100°C and cool down to ~37°C.</li>
 +
  <li>Add E. coli (1x10<sup>9</sup> cells/ml) expressing GFP in the 1% alginate.</li>
 +
  <li>Make 2% CaCl<sub>2</sub>: 10 g of Calcium chloride (SIGMA-ALDRICH) in 500 ml of H<sub>2</sub>O</li>
 +
  <li>Use a dropper to aspirate the 1% alginate containing bacteria and drop slowly into 2% CaCl<sub>2</sub>.</li>
 +
  <li>Incubate for 30 seconds and pick up the micro-spheroid particles carrying bacteria.</li>
 +
  </ol>
 +
  </h4>
 +
  <div class="row">
 +
 +
  <div class="col-md-6 col-sm-6">
 +
  <img src="https://static.igem.org/mediawiki/2017/8/83/Mingdao_igem_ethan_pic1.png" width="100%">
 +
  </div>
 +
      <div class="col-md-6 col-sm-6">
 +
  <img src="https://static.igem.org/mediawiki/2017/f/fe/Mingdao_igem_ethan_pic2.png" width="100%">
 +
  </div>
 +
 +
  <div class="col-md-2 col-sm-2">&nbsp;</div>
 +
  <div class="col-md-8 col-sm-8">
 +
  <video width="100%" style="padding-top: 2rem" controls>
 +
<source src="https://static.igem.org/mediawiki/2017/b/b7/Mingdao_igem_ethan_micro2.mp4" type="video/mp4">
 +
</video>
 +
  </div>
 +
  <div class="col-md-2 col-sm-2">&nbsp;</div>
 +
  </div>
 +
  <h3 style="padding-top: 2rem"><b>Test in Acidic and Basic Environment</b></h3>
 +
<h4 class="w3-padding-24">
 +
  <ol>
 +
  <li>Make solutions of pH=7, pH = 2, pH=9 to mimic esophagus stomach intestine environment, respectively.</li>
 +
  <li>Put ~30 micro-spheroid particles carrying bacteria into 50ml of each solution</li>
 +
  <li>Keep at room temperature for 4 hours</li>
 +
  <li>Take 100ul to one well of 96-well plate and measure GFP at Ex/Em=485/528nm with a BioTek microplate reader.</li>
 +
  <li>Spread 100ul onto agar plates. Grow at 37°C overnight and count the numbers of colonies.</li>
 +
  </ol>
 +
  </h4>
 +
  <div class="row">
 +
  <div class="col-md-2 col-sm-2">&nbsp;</div>
 +
<div class="col-md-8 col-sm-8">
 +
<video width="100%" style="padding-top: 2rem" controls>
 +
  <source src="https://static.igem.org/mediawiki/2017/5/57/Ethan_mingdao_igem_micro1.mp4" type="video/mp4">
 +
  </video>
 +
</div>
 +
<div class="col-md-2 col-sm-2">&nbsp;</div>
 +
  </div>
 +
 +
<h3><b>Results</b></h3>
 +
<div class="row">
 +
<div class="col-md-6 col-sm-6">
 +
<img src="https://static.igem.org/mediawiki/2017/3/36/Mingdao_igem_ethan_pic3.png" width="100%">
 +
<h4><i class="fa fa-arrow-circle-up" aria-hidden="true"></i>&nbsp;&nbsp;The GFP values are higher in pH=7 and pH=9 compared to pH=2.</h4>
 +
</div>
 +
<div class="col-md-6 col-sm-6">
 +
<img src="https://static.igem.org/mediawiki/2017/f/fa/Mingdao_igem_ethan_pic4.png" width="100%">
 +
<h4><i class="fa fa-arrow-circle-up" aria-hidden="true"></i>&nbsp;&nbsp;1071 colonies grew on agar plates from the solution of pH=9. But there were not any colonies surviving from the solution of pH=2</h4>
 +
</div>
 +
</div>
 +
  <h4 class="w3-padding-24">
 +
</h4>
 +
    <h3 class="w3-padding-24" style="padding: 0 3rem 3rem 3rem">
 +
  The data above indicated that the micro-spheroid particles may release the microbes which survive in an environment of pH=7~9.
 +
  </h3>
 +
  <!-- PART end -->
 +
  <!-- REFERENCE start -->
 +
 +
                  <section id="expert">
 +
                      <div style="margin-left: 20px">
 +
                          <!-- Box -->
 +
                          <div class="box" style="background-color: rgba(255, 255, 255, 0); margin-right: 50px">
 +
                              <ul class="list-group">
 +
                                <li class="list-group-item" style="text-align: center; font-size: 1.3rem">Reference</li>
 +
                                <li class="list-group-item">Preparation methods and applications behind alginate-based particles. Expert Opin Drug Deliv. 2017;14(6):769-782.</li>
 +
                              </ul>
 +
                          </div>
 +
                      </div>
 +
                  </section>
 +
                  <!-- REFERENCE end-->
 +
</div>
 +
</div>
 
                 <div class="card" style="margin-bottom: 3rem">
 
                 <div class="card" style="margin-bottom: 3rem">
 
                       <div class="card-header text-center bg-info" style="color: white; letter-spacing: 0.3rem;font-size:1.8rem;">
 
                       <div class="card-header text-center bg-info" style="color: white; letter-spacing: 0.3rem;font-size:1.8rem;">
                         Framework
+
                         Simulation With Prototype
 
                       </div>
 
                       </div>
 
                       <div class="card-body">
 
                       <div class="card-body">
 
                           <!-- PART start -->
 
                           <!-- PART start -->
 +
  <div class="row">
 +
  <div class="col-md-2 col-sm-2">&nbsp;</div>
 +
  <div class="col-md-8 col-sm-8">
 +
  <video width="100%" controls>
 +
  <source src="https://static.igem.org/mediawiki/2017/5/58/Mingdaojay1031-1.mp4" type="video/mp4">
 +
</video>
 +
  </div>
 +
  <div class="col-md-2 col-sm-2">&nbsp;</div>
 +
  </div>
 
                           <h4 class="w3-padding-24">
 
                           <h4 class="w3-padding-24">
                               We  designed  a  cable  car-liked   device  to  simulate   the  situation  of   food  (pills)  entering human  body.  By  the  environment  it  has  to  stays  like  mouth,  stomach,  i  ntestinal   tract,  and how  long  it  will  stay,  we  ensure   that  our  pills   will  be  released  only  in  intestinal  tract  rather than  break   down  in  inappropriate   position   (Too  early  or  too  late).  What’s  more,  this  device  is not   only   designed  for  our  project,   it   can  be  used  to  test  any  other  kind   of  pills   easily  with simply  changing  the  substance   in  the  liquid  and  the  operating  time  of  the  cable   car.
+
                               We  designed  a  cable  car-like   device  to  simulate  food  (capsule)  entering   and  passing  through  the  human  digestive  system.  By  the  environment  it  has  to  stays  like  mouth,  stomach,  intestinal   tract,  and   how  long  it  will  stay,  we  ensured   that  our  capsule   will  be  released  only  in   the   intestinal  tract  rather than  breaking   down  in  unintended   areas   (Too  early  or  too  late).  What’s  more,  this  device  is   not  designed   only   for  our  project.   It   can  be  used  to  test  any  other  kinds   of  capsule   easily  by  simply  changing  the  substances   in  the  liquid  and  the  operating  time  of  the  cable car.
 
                           </h4>
 
                           </h4>
 
                           <!-- PART end -->
 
                           <!-- PART end -->
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                               <!-- PART start -->
 
                               <!-- PART start -->
 
                               <h4 class="w3-padding-24">
 
                               <h4 class="w3-padding-24">
                                   We   make   Acrylic   Sheet   a   box   and   a   cover   with   holes.   The   container   is   separated   into three   sections──   neutral(mouth),   Acid(stomach),   and   Alkaline(intestinal   tract).   The   holes   on the   cover   are   to   avoid   the   liquid   inside   splitting   out,   but   the   size   of   them   are   designed   to   let   a pill  get   through   exactly.
+
                                   We make Acrylic Sheet a box and a cover with holes. The container is separated into three sections── neutral(mouth), Acid(stomach), and Alkaline(intestinal tract). The holes on the cover are to avoid the liquid inside splitting out, but the size of them are designed to let a capsule get through exactly.
 
                               </h4>
 
                               </h4>
 
                               <img src="https://static.igem.org/mediawiki/2017/4/4f/Mingdaochiwen_1020-2.png" width="80%" style="display: block; margin: auto">
 
                               <img src="https://static.igem.org/mediawiki/2017/4/4f/Mingdaochiwen_1020-2.png" width="80%" style="display: block; margin: auto">
                               <h4 style="text-align: center; letter-spacing: 0.1rem"><b>Figure  2   NXT   Lego  track  device</b></h4>
+
                               <h4 style="text-align: center; letter-spacing: 0.1rem"><b>Figure  2   NXT LEGO track  device</b></h4>
  
 
                               <h4 class="w3-padding-24">
 
                               <h4 class="w3-padding-24">
                                   We   chose   NXT   Lego   to   build   the   cable   car   track   above   because   of   its   simplicity   and creative.   Put   the   cable   car-liked   device   on   the   cover   and   start   the   program.Soaking   the   pills in   the   liquid   orderly,   and   decide   how   long   we   soak   the   pills  according   to   the   time   food   stays in   the   Digestive   tract.
+
                                   We chose NXT Lego to build the cable car track above because of its simplicity and creative. Put the cable car-liked device on the cover and start the program.Soaking the capsule in the liquid orderly, and decide how long we soak the capsule according to the time food stays in the Digestive tract.
 
                               </h4>
 
                               </h4>
  
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                             <h4 style="text-align: center; letter-spacing: 0.1rem"><b>Figure  4  Container</b></h4>
 
                             <h4 style="text-align: center; letter-spacing: 0.1rem"><b>Figure  4  Container</b></h4>
  
                                 </div>
+
                        </div>
 +
<div class="card" style="margin-bottom: 3rem">
 +
                              <div class="card-header text-center" style="color: white; letter-spacing: 0.3rem;font-size:1.8rem; background-color:#1DC7D7">
 +
                                 User Guide
 +
                            </div>
 +
                            <h4 style="text-align: center; letter-spacing: 0.1rem; padding: 3rem 3rem 3rem 3rem ">Our cable car-liked device is simple and safe for anyone to use. User only have to do is hang the capsule which waiting for detection on the transport carriage, and push the start button on the core(the orange one). Then observe the result patiently. It can show us when will capsule break down and release medicine. It can also test various pill by just adjust the solution in the container or the time the capsule soak in solution.</h4>
 +
<img src="https://static.igem.org/mediawiki/2017/6/6b/Mingdaochiwen_1031-1.png" width="80%" style="display: block; margin: auto">
 +
                        </div>
 
                             </div>
 
                             </div>
  

Latest revision as of 01:21, 2 November 2017

iGEM Human Practice



Prototype

Not only in the lab,
we brought our design into reality


Acid-resistant Microparticles

To make probiotics as a good food supplement, one may consider the engineered microbes need to pass through gastric acid in the digestive system. Calcium alginate nanoparticles are good carriers for delivery which are acid-resistant (pH=2) and releasing in basic environment (pH=9). The following is our protocol in the microencapsulation process.

Microencapsulation

  1. Make 0.25% alginate: 0.25 g of Alginic acid sodium salt (SIGMA-ALDRICH) in 100 ml of H2O. Heat at 100°C and cool down to ~37°C.
  2. Add E. coli (1x109 cells/ml) expressing GFP in the 1% alginate.
  3. Make 2% CaCl2: 10 g of Calcium chloride (SIGMA-ALDRICH) in 500 ml of H2O
  4. Use a dropper to aspirate the 1% alginate containing bacteria and drop slowly into 2% CaCl2.
  5. Incubate for 30 seconds and pick up the micro-spheroid particles carrying bacteria.

 
 

Test in Acidic and Basic Environment

  1. Make solutions of pH=7, pH = 2, pH=9 to mimic esophagus stomach intestine environment, respectively.
  2. Put ~30 micro-spheroid particles carrying bacteria into 50ml of each solution
  3. Keep at room temperature for 4 hours
  4. Take 100ul to one well of 96-well plate and measure GFP at Ex/Em=485/528nm with a BioTek microplate reader.
  5. Spread 100ul onto agar plates. Grow at 37°C overnight and count the numbers of colonies.

 
 

Results

  The GFP values are higher in pH=7 and pH=9 compared to pH=2.

  1071 colonies grew on agar plates from the solution of pH=9. But there were not any colonies surviving from the solution of pH=2

The data above indicated that the micro-spheroid particles may release the microbes which survive in an environment of pH=7~9.

  • Reference
  • Preparation methods and applications behind alginate-based particles. Expert Opin Drug Deliv. 2017;14(6):769-782.
Simulation With Prototype
 
 

We designed a cable car-like device to simulate food (capsule) entering and passing through the human digestive system. By the environment it has to stays like mouth, stomach, intestinal tract, and how long it will stay, we ensured that our capsule will be released only in the intestinal tract rather than breaking down in unintended areas (Too early or too late). What’s more, this device is not designed only for our project. It can be used to test any other kinds of capsule easily by simply changing the substances in the liquid and the operating time of the cable car.

Design

Figure 1 Liquid container

We make Acrylic Sheet a box and a cover with holes. The container is separated into three sections── neutral(mouth), Acid(stomach), and Alkaline(intestinal tract). The holes on the cover are to avoid the liquid inside splitting out, but the size of them are designed to let a capsule get through exactly.

Figure 2 NXT LEGO track device

We chose NXT Lego to build the cable car track above because of its simplicity and creative. Put the cable car-liked device on the cover and start the program.Soaking the capsule in the liquid orderly, and decide how long we soak the capsule according to the time food stays in the Digestive tract.

Component

Figure 3 Lid of container

Figure 4 Container

User Guide

Our cable car-liked device is simple and safe for anyone to use. User only have to do is hang the capsule which waiting for detection on the transport carriage, and push the start button on the core(the orange one). Then observe the result patiently. It can show us when will capsule break down and release medicine. It can also test various pill by just adjust the solution in the container or the time the capsule soak in solution.

iGEM Human Practice

  Human Practice

Gold

Silver

Collaborate

Engagement

Achievements