Team:Valencia/co-culter

From 2012.igem.org

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Our furthest goal is to build a continuous culture system with spatial and temporal decoupling from the photosynthetic and the bioluminescent module, totally autonomous. <br><br>
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Our furthest goal is to build a continuous culture system with spatial and temporal decoupling from the photosynthetic and the bioluminescent module, totally autonomous.  
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<br><br>
<h3><u>Method:</u></h3><br>
<h3><u>Method:</u></h3><br>
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Set 2 separate culture modules, a flat wide one for <i>S. elongatus cscB</i> as a solar module and a smaller compact one for <i>A. fischeri</i> as a biobulb. Prepare an open system so that there can be gas exchange from the cultures with the atmosphere (to let the system at as a CO<sub>2</sub> sink), but with a Pasteurian design opening to avoid contamination from deposition.<br><br>
+
Set 2 separate culture modules, a flat wide one for <i>S. elongatus cscB</i> as a solar module and a smaller compact one for <i>A. fischeri</i> as a biobulb. Prepare an open system so that there can be gas exchange from the cultures with the atmosphere (to let the system at as a CO<sub>2</sub> sink), but with a Pasteurian design opening to avoid contamination from deposition.
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Set tubing connecting both cultures, protecting each end with a syringe filter 0.45 microns of pore size. The fluid medium dynamics is borne by a pair of peristaltic reversible-flow pumps, which switch flow direction every 30-60 seconds to avoid collapsing any side of the membranes with jammed cells. The membranes allow the exchange of gases, ions, water, sucrose and AHL, but not cells, so that populations do not mix. This is fundamental to guarantee the light-efficiency of each compartment, without having cyanobacteria shading light emission from <i>A. fischeri</i>, and permitting a spatial decoupling of the photosynthetic module and the biolamp.<br><br>
+
<br><br>
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Cell density would be regulated by output tubes from each culture which would recycle broth after passing it through a filter/skimmer. This process would be regulated by a turbidimeter (in theory) or by sampling-and-testing, at least in the first experimental prototype.<br><br>
+
Set tubing connecting both cultures, protecting each end with a syringe filter 0.45 microns of pore size. The fluid medium dynamics is borne by a pair of peristaltic reversible-flow pumps, which switch flow direction every 30-60 seconds to avoid collapsing any side of the membranes with jammed cells. The membranes allow the exchange of gases, ions, water, sucrose and AHL, but not cells, so that populations do not mix. This is fundamental to guarantee the light-efficiency of each compartment, without having cyanobacteria shading light emission from <i>A. fischeri</i>, and permitting a spatial decoupling of the photosynthetic module and the biolamp.
-
Water loss due to evaporation would be solved by a water-level-wise lever opening a valve which would let in some distilled water from a small deposit, when the volume of the liquid falls below certain threshold.<br><br>
+
<br><br>
-
A microcontroller would be used as hardware to coordinate pump flow switch, input signals from turbidimeters and control of the cell density control system.<br><br>
+
Cell density would be regulated by output tubes from each culture which would recycle broth after passing it through a filter/skimmer. This process would be regulated by a turbidimeter (in theory) or by sampling-and-testing, at least in the first experimental prototype.
-
All electric devices would be powered by a small solar panel, to preserve to the last detail the energetic autonomy of the system.<br><br>
+
<br><br>
-
Routine analyses of the system would be carried out on sucrose, oxygen and AHL levels in the liquid medium. Luminescence would be measured at night to acquire an experimental Light/Time curve.<br><br>
+
Water loss due to evaporation would be solved by a water-level-wise lever opening a valve which would let in some distilled water from a small deposit, when the volume of the liquid falls below certain threshold.
-
[Future design development: Achieve a knock-out/directed mutagenesis of the luxI autoinducer genes in <i>A. fischeri</i> to unable isolated autoinduction. This would render the system totally dependent on the AHL production of the transformed <i>S. elongatus</i>. In such situation, turning valves of different membrane pore size at the cell-stopping sections would result extremely useful, to manually switch on/off the diffusion of AHL from the <i>Synechococcus</i> culture to the biolamp. This enables an element of human control to turn on/off the lights over the normal day/night fashion].<br><br>
+
<br><br>
 +
A microcontroller would be used as hardware to coordinate pump flow switch, input signals from turbidimeters and control of the cell density control system.
 +
<br><br>
 +
All electric devices would be powered by a small solar panel, to preserve to the last detail the energetic autonomy of the system.
 +
<br><br>
 +
Routine analyses of the system would be carried out on sucrose, oxygen and AHL levels in the liquid medium. Luminescence would be measured at night to acquire an experimental Light/Time curve.
 +
<br><br>
 +
[Future design development: Achieve a knock-out/directed mutagenesis of the luxI autoinducer genes in <i>A. fischeri</i> to unable isolated autoinduction. This would render the system totally dependent on the AHL production of the transformed <i>S. elongatus</i>. In such situation, turning valves of different membrane pore size at the cell-stopping sections would result extremely useful, to manually switch on/off the diffusion of AHL from the <i>Synechococcus</i> culture to the biolamp. This enables an element of human control to turn on/off the lights over the normal day/night fashion].
 +
<br><br>
<center>
<center>
<img src="http://2012.igem.org/wiki/images/5/58/Advanced_coculture_vlc_BBFJFJBFJBFJ.JPG">
<img src="http://2012.igem.org/wiki/images/5/58/Advanced_coculture_vlc_BBFJFJBFJBFJ.JPG">

Revision as of 11:20, 26 September 2012



Advanced continuous coculture system


Our furthest goal is to build a continuous culture system with spatial and temporal decoupling from the photosynthetic and the bioluminescent module, totally autonomous.

Method:


Set 2 separate culture modules, a flat wide one for S. elongatus cscB as a solar module and a smaller compact one for A. fischeri as a biobulb. Prepare an open system so that there can be gas exchange from the cultures with the atmosphere (to let the system at as a CO2 sink), but with a Pasteurian design opening to avoid contamination from deposition.

Set tubing connecting both cultures, protecting each end with a syringe filter 0.45 microns of pore size. The fluid medium dynamics is borne by a pair of peristaltic reversible-flow pumps, which switch flow direction every 30-60 seconds to avoid collapsing any side of the membranes with jammed cells. The membranes allow the exchange of gases, ions, water, sucrose and AHL, but not cells, so that populations do not mix. This is fundamental to guarantee the light-efficiency of each compartment, without having cyanobacteria shading light emission from A. fischeri, and permitting a spatial decoupling of the photosynthetic module and the biolamp.

Cell density would be regulated by output tubes from each culture which would recycle broth after passing it through a filter/skimmer. This process would be regulated by a turbidimeter (in theory) or by sampling-and-testing, at least in the first experimental prototype.

Water loss due to evaporation would be solved by a water-level-wise lever opening a valve which would let in some distilled water from a small deposit, when the volume of the liquid falls below certain threshold.

A microcontroller would be used as hardware to coordinate pump flow switch, input signals from turbidimeters and control of the cell density control system.

All electric devices would be powered by a small solar panel, to preserve to the last detail the energetic autonomy of the system.

Routine analyses of the system would be carried out on sucrose, oxygen and AHL levels in the liquid medium. Luminescence would be measured at night to acquire an experimental Light/Time curve.

[Future design development: Achieve a knock-out/directed mutagenesis of the luxI autoinducer genes in A. fischeri to unable isolated autoinduction. This would render the system totally dependent on the AHL production of the transformed S. elongatus. In such situation, turning valves of different membrane pore size at the cell-stopping sections would result extremely useful, to manually switch on/off the diffusion of AHL from the Synechococcus culture to the biolamp. This enables an element of human control to turn on/off the lights over the normal day/night fashion].



Results:


Construction of inter-flask pumping system with 0.45micron pore membranes.