Difference between revisions of "Team:CU Boulder/Design"
Line 2: | Line 2: | ||
<html lang="en"> | <html lang="en"> | ||
<head> | <head> | ||
+ | <meta content="text/html; charset=ISO-8859-1" | ||
+ | http-equiv="content-type"> | ||
+ | <title>Design</title> | ||
</head> | </head> | ||
<body> | <body> | ||
− | < | + | <br> |
− | + | <article> | |
− | <h1> | + | <h1 style="text-align: center;">The BioStake</h1> |
− | + | <div style="text-align: center;"><img | |
− | + | style="width: 398px; height: 398px;" alt="CU iGEM BioStake" | |
− | + | src="https://static.igem.org/mediawiki/2015/8/8d/Igem_stake.gif"></div> | |
− | + | <big><big><br> | |
− | + | <br> | |
− | < | + | The general design of the stake was inspired <a target="_blank" |
+ | href="http://onlinelibrary.wiley.com/doi/10.1021/bp9600142/abstract">this | ||
+ | paper</a> published in <span style="font-style: italic;">Biotechnology | ||
+ | Progress</span> which describes a miniature bioreactor to detect | ||
+ | toxicity by using bioluminescent E. coli. Our design, shown in the | ||
+ | image, will holds cells in a gel at the top of the long | ||
+ | ground-insertion portion and will include homeostatic controls on the | ||
+ | top of the stake. The outer filtration and pump aspects are not shown | ||
+ | in the design above, but will be included to protect the cells from | ||
+ | harmful environmental influences.</article></big></big><br> | ||
+ | <br> | ||
+ | <br> | ||
<br> | <br> | ||
<h2><big>Biological Design Considerations</big></h2> | <h2><big>Biological Design Considerations</big></h2> | ||
Line 32: | Line 46: | ||
relatively fixed position, and its biodegradability, which is | relatively fixed position, and its biodegradability, which is | ||
attractive in the context of making an environmentally-conscious | attractive in the context of making an environmentally-conscious | ||
− | product.<br> | + | product. Additionally, by developing a hydrogel with relatively small |
+ | pore size such as with PVA, it can act as a filter to keep out larger | ||
+ | competetive bacteria that may infiltrate the device as well as prevent | ||
+ | solid sedimentation from blocking cell-to-cell interaction and nutrient | ||
+ | diffusion.<br> | ||
<br> | <br> | ||
− | In early September, PVA scaffolds were made which contained cells with | + | </big></big><big><big>In early September, PVA |
− | the pBad inducible promoter in front of the RFP gene, and | + | scaffolds were made which contained |
− | results support the hypothesis that the engineered E. coli were able to | + | cells with the pBad inducible promoter in front of the RFP gene, and |
− | live inside and respond to arabinose when soaked in the solution. | + | experimental results support the hypothesis that the engineered E. coli |
− | </big></big><big><span style="font- | + | were able to live inside and respond to arabinose when soaked in the |
+ | solution. Further experimentation described below is still necessary to | ||
+ | confirm that this </big></big><br> | ||
+ | <big><big><br> | ||
+ | In addition to designing the spatial arrangment of the cells, it's also | ||
+ | important to consider elements that will allow for their survival over | ||
+ | the time period which the stake is in place to collect data. If <span | ||
+ | style="font-style: italic;">E. coli </span>cells are | ||
+ | not kept in highly sterile conditions within a narrow temperature, | ||
+ | humidity, and nutrient concentration range, they will not sustain a | ||
+ | functional cell count for the system to work properly. Therefore, a | ||
+ | control system that can sense and automatically adjust these conditions | ||
+ | must be incorporated. The proposed plan is to use a solar-powered stake | ||
+ | so batteries do not have to be replaced, creating a more sustainable | ||
+ | device that can be used repeadetly. However, the incorporation of such | ||
+ | technology could potentially drive up the cost of the BioStake which | ||
+ | would not be favorable considering its intended use is for quick, easy, | ||
+ | and low-cost results available to anybody. Nonetheless, it seems to be | ||
+ | a | ||
+ | feasible solution to work with the cells that we have already developed.<br> | ||
<br> | <br> | ||
+ | <br> | ||
+ | </big></big><big><span style="font-weight: bold;"></span></big><big><span | ||
+ | style="font-weight: bold;"><br> | ||
</span></big> | </span></big> | ||
<h2><big>Designing for Environmental Functionality</big></h2> | <h2><big>Designing for Environmental Functionality</big></h2> | ||
+ | <big><big>The types of locations in which the BioStake will | ||
+ | be placed, and how it tracks contamination at the sites is crucial to | ||
+ | the design of the stake itself. To create the best device, it should be | ||
+ | able | ||
+ | to intake an air or water sample that is consistent with the | ||
+ | bioavaiable concentrations of fracking contaminants, while excluding | ||
+ | harmful components from affecting the bacteria. The BioStake would | ||
+ | contain a small pump that would attract water or air through a filter | ||
+ | wraped around the base of the stake, and then small tubes would draw up | ||
+ | the fluid to the cell-containing gel via capillary action. | ||
+ | </big></big><br> | ||
<br> | <br> | ||
<br> | <br> | ||
<br> | <br> | ||
<h2><big>Creating a User-Friendly Device</big></h2> | <h2><big>Creating a User-Friendly Device</big></h2> | ||
− | <big>< | + | <big><big>As described on the motivation page, the market |
− | <br> | + | has high demand for a biosensor that is easy to use and available to |
+ | the general public to detect fracking contaminants. Apart from the | ||
+ | design of the pump and homeostasis mechanism, many components of the | ||
+ | stake are designed specifically to make the BioStake inexpensive and | ||
+ | easy-to-use. <br> | ||
<br> | <br> | ||
+ | The physical structure of the stake is intended to be simple enough | ||
+ | that the main components could be manufactured in a plastic injection | ||
+ | mold. It's also designed for the hydrogel to be held in the compartment | ||
+ | between the top and the long ground-insertion portion so that the user | ||
+ | can know whether or not napthalene is present based on visual | ||
+ | inspection of the device to see whether the gel has changed colors. The | ||
+ | stake itself would be less than 6 inches in length and would weigh less | ||
+ | than a pound. Research still has to be done to balance being small | ||
+ | enough to be portable, and long enough for it to be able to collect | ||
+ | continuous fluid samples from air, water, or soil in the environments | ||
+ | around fracking sites.<br> | ||
+ | </big></big><big><br> | ||
</big> | </big> | ||
<h2><big>Future Directions for Development</big></h2> | <h2><big>Future Directions for Development</big></h2> | ||
Line 54: | Line 121: | ||
<big><big>Further experimentation is required to determine | <big><big>Further experimentation is required to determine | ||
the | the | ||
− | feasibility of AHL diffusion in cell-to-cell signaling, extensive cell | + | feasibility of using PVA to allow AHL diffusion in cell-to-cell |
− | growth studies to see if the system, and | + | signaling, extensive cell |
+ | growth studies to see if the system can continue. If we had more time, | ||
+ | measurements would be taken to determine cell growth rates over time, | ||
+ | and then the hydrogel chemistry could be adjusted to control nutrient | ||
+ | diffusion appropriately.<br> | ||
<br> | <br> | ||
+ | Additionally, the | ||
+ | proposed method to maintain homeostasis within the device that would | ||
+ | allow the cells to survive over long periods of time may be tedious to | ||
+ | maintain and the aspect of the design that is most vulnerable to | ||
+ | failure. It may be beneficial to move the design from <span | ||
+ | style="font-style: italic;">E. coli</span> into a | ||
+ | bacterium that can survive in much harsher conditions, perhaps a | ||
+ | bacteria that already thrives in natural water sources.</big></big><br> | ||
<br> | <br> | ||
+ | <big><big>Ideally, this product could be prototyped and put | ||
+ | into mass production. However, without enough information to specify | ||
+ | exact design criteria, its difficult to develop an entrepreneurial | ||
+ | plan, despite environmental agencies and oil companies expressing | ||
+ | interest in the product. </big></big><br> | ||
<br> | <br> | ||
<br> | <br> | ||
<br> | <br> | ||
+ | <br> | ||
+ | <br> | ||
+ | <br> | ||
+ | <br> | ||
+ | | ||
</article> | </article> | ||
</body> | </body> | ||
</html> | </html> | ||
+ | |||
{{CU_Boulder_footer}} | {{CU_Boulder_footer}} |
Revision as of 13:10, 17 September 2015
<!DOCTYPE html>