Difference between revisions of "Team:Bielefeld-CeBiTec/Results/it-really-works"
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| − | <figure style="width: 350px; margin-right: 20px; float: left"> | + | <figure style="width: 350px; margin-right: 20px; margin-bottom: 20px; float: left;"> |
<a href="https://static.igem.org/mediawiki/2015/0/08/Bielefeld-CeBiTec_prototype_box_and_smartphone_up_big.png" data-lightbox="finalaim" data-title="Don't drink this water!"><img src="https://static.igem.org/mediawiki/2015/7/71/Bielefeld-CeBiTec_prototype_box_and_smartphone_up_small.png" alt="box"></a> | <a href="https://static.igem.org/mediawiki/2015/0/08/Bielefeld-CeBiTec_prototype_box_and_smartphone_up_big.png" data-lightbox="finalaim" data-title="Don't drink this water!"><img src="https://static.igem.org/mediawiki/2015/7/71/Bielefeld-CeBiTec_prototype_box_and_smartphone_up_small.png" alt="box"></a> | ||
| − | + | </figure> | |
| − | + | <figure style="width: 350px; margin-right: 20px; float: left;"> | |
<a href="https://static.igem.org/mediawiki/2015/f/f6/Bielefeld-CeBiTec_all_in_one_box_seite_big.png" data-lightbox="finalaim" data-title="Prototype box for detection"><img src="https://static.igem.org/mediawiki/2015/c/c3/Bielefeld-CeBiTec_all_in_one_box_seite_small.png" alt="Device box"></a> | <a href="https://static.igem.org/mediawiki/2015/f/f6/Bielefeld-CeBiTec_all_in_one_box_seite_big.png" data-lightbox="finalaim" data-title="Prototype box for detection"><img src="https://static.igem.org/mediawiki/2015/c/c3/Bielefeld-CeBiTec_all_in_one_box_seite_small.png" alt="Device box"></a> | ||
<figcaption> Our measurement prototype for easy and fast evaluation of fluorescence emission </figcaption> | <figcaption> Our measurement prototype for easy and fast evaluation of fluorescence emission </figcaption> | ||
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| − | <p>We aimed at developing a cell-free test strip for the detection of various substances at once. We started with many different | + | <p>We aimed at developing a cell-free test strip for the parallel detection of various substances at once. We started with many different subprojects. A lot of cloning needed to be performed, the ideal design of the teststrip had to be developed, a cell-free detection approach had to be established and the processing of the output signal had to be taken care of. So in our last weeks our main focus was to bring all this different aspects together to one working approach. And we did it successfully! </p> |
| − | <p> We achieved | + | <p> We achieved a lot during this project, just take a look at this (far from complete) list to get an idea:</p> |
<ol> | <ol> | ||
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| − | <video muted controls src = "https://static.igem.org/mediawiki/2015/8/84/Bielefeld-CeBiTec_AllinOneVideo.mp4" type="video/mp4" width="1060" | + | |
| + | |||
| + | <h2>Heavy Metal Detection in Detail</h2> | ||
| + | <p> We made it possible to detect various heavy metals with our sensor. The sensor is based on standard ligand-transcription factor interaction. We tested our sensor by using mercury and copper contaminated water. The water sample was applied to the cell-free paper teststrip, which contained the freeze dried cell-free protein synthesis (CFPS) reaction. Our sensor was able to discriminate between clean and contaminated water. This result was obtained by taking a picture of the strip with a common smartphone. The only thing one needs is our black case, filters in front of your flash and camera as well as our self-programmed app. The app quantifies the fluorescence in real-time and calculates whether a contamination is present in the sample or not. A typical workflow is depicted in this video:</p> | ||
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| + | <video muted controls src = "https://static.igem.org/mediawiki/2015/8/84/Bielefeld-CeBiTec_AllinOneVideo.mp4" type="video/mp4" width="1060" | ||
height="525" | height="525" | ||
poster="https://static.igem.org/mediawiki/2015/0/08/Bielefeld-CeBiTec_prototype_box_and_smartphone_up_big.png"> | poster="https://static.igem.org/mediawiki/2015/0/08/Bielefeld-CeBiTec_prototype_box_and_smartphone_up_big.png"> | ||
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</video> | </video> | ||
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<div class="col-md-12"> | <div class="col-md-12"> | ||
| + | <h2>Exemplary wet lab detection of Mercury</h2> | ||
| + | |||
| + | <p> At molecular level, all our biosensors work on basis of the simple transcription factor-ligand interaction concept: The expression of a gene is regulated by a transcription factor protein. In case of a transcriptional repressor, the reporter protein (sfGFP) is not expressed unless a particular ligand is present and changes the conformation of the transcription factor. In case of a transcriptional activator, expression of the reporter protein is enhanced in presence of the ligand. </p> | ||
| + | <p>This regulation is a central process in nature. Because our system uses this concept, it is <b>highly extensible</b> and can be used to tackle various real world problems.</p> | ||
| + | |||
<figure style="width: 450px; margin-right: 20px; float: left"> | <figure style="width: 450px; margin-right: 20px; float: left"> | ||
<a href="https://static.igem.org/mediawiki/2015/e/e8/Bielefeld-CeBiTec_all_in_one_hg_without_filter_big.png" data-lightbox="finalaim" data-title="Cell-free biosensor for mercury detection in water samples, pictured without filters."><img src="https://static.igem.org/mediawiki/2015/6/66/Bielefeld-CeBiTec_all_in_one_hg_without_filter_small.png" alt="Cell free Hg sensor"></a> | <a href="https://static.igem.org/mediawiki/2015/e/e8/Bielefeld-CeBiTec_all_in_one_hg_without_filter_big.png" data-lightbox="finalaim" data-title="Cell-free biosensor for mercury detection in water samples, pictured without filters."><img src="https://static.igem.org/mediawiki/2015/6/66/Bielefeld-CeBiTec_all_in_one_hg_without_filter_small.png" alt="Cell free Hg sensor"></a> | ||
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| − | + | <p></p> | |
<p> We consider our cell-free biosensor an outstanding instrument for the detection of harmful substances in liquids. </p> | <p> We consider our cell-free biosensor an outstanding instrument for the detection of harmful substances in liquids. </p> | ||
<p> We see great need for a storable, transportable and easy-to-use water quality test strip all around the world.</p> | <p> We see great need for a storable, transportable and easy-to-use water quality test strip all around the world.</p> | ||
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| − | <p> During the reaction we had measured the fluorescence in a plate reader. In the nearby figures you can see: The standard <a data-toggle="tooltip" title="Cell-Free Protein Synthesis">CFPS</a> setup performed not sufficent when mercury was present, HOWEVER, when using our mercury detection setup, cell-free synthesis of our reporter protein sfGFP was greatly enhanced! </p> | + | <p> During the reaction we had measured the fluorescence in a plate reader, in order to verify the results obtained by the app. In the nearby figures you can see: The standard <a data-toggle="tooltip" title="Cell-Free Protein Synthesis">CFPS</a> setup performed not sufficent when mercury was present, HOWEVER, when using our mercury detection setup, cell-free synthesis of our reporter protein sfGFP was greatly enhanced! </p> |
<figure style="width: 500px; margin-right: 20px; float: left"> | <figure style="width: 500px; margin-right: 20px; float: left"> | ||
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<div class="col-md-12"> | <div class="col-md-12"> | ||
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| − | < | + | <h2>Detection of Date Rape Drugs</h2> |
| − | + | <p> With our developed biosensor we <b>introduce an easy and unprecedented date rape drug ingredient biosensor!</b> To detect the date rape drug ingredient γ-hydroxy butyrate (GHB), we had to adjust <b>only two components</b> of our setup: The repressor protein and the DNA sequence that the protein recognizes. </p> | |
| − | <p> | + | |
Revision as of 03:53, 19 September 2015
All-in-One Performance
It LITERALLY works on paper!
Project aim: Achieved!
We aimed at developing a cell-free test strip for the parallel detection of various substances at once. We started with many different subprojects. A lot of cloning needed to be performed, the ideal design of the teststrip had to be developed, a cell-free detection approach had to be established and the processing of the output signal had to be taken care of. So in our last weeks our main focus was to bring all this different aspects together to one working approach. And we did it successfully!
We achieved a lot during this project, just take a look at this (far from complete) list to get an idea:
- Highly efficient and robust cell-free synthesis of sfGFP on simple paper that can be stored. By the way, you just have to spend 16 ¢ for each reaction
- Cloning and characterization of multiple heavy metal sensors
- Cloning and characterization of a date rape drug sensor
- Establishment of a smartphone based fluorescence detection method
- Implementing of an app to easily evaluate the measured fluorescence signals.
So this is, how they actually work together!
- At first, you have to combine all reagents that are necessary to conduct cell-free protein synthesis.
- When all compounds are combined, the reaction is applied on paper and freeze-dried. This procedure prevents the reaction from premature starting. Additionally, all remaining cells are killed.
- The paper with the reaction is storable and transportable. For examination of water quality, you can simply add a water drop to the paper.
- In no time you can take a picture of the paper with your smartphone. A filter combination for your smartphone enables you to capture the fluorescence that is emitted by the reporter protein.
- Our app evaluates the data and tells you if your water sample is contaminated.
Heavy Metal Detection in Detail
We made it possible to detect various heavy metals with our sensor. The sensor is based on standard ligand-transcription factor interaction. We tested our sensor by using mercury and copper contaminated water. The water sample was applied to the cell-free paper teststrip, which contained the freeze dried cell-free protein synthesis (CFPS) reaction. Our sensor was able to discriminate between clean and contaminated water. This result was obtained by taking a picture of the strip with a common smartphone. The only thing one needs is our black case, filters in front of your flash and camera as well as our self-programmed app. The app quantifies the fluorescence in real-time and calculates whether a contamination is present in the sample or not. A typical workflow is depicted in this video:
Exemplary wet lab detection of Mercury
At molecular level, all our biosensors work on basis of the simple transcription factor-ligand interaction concept: The expression of a gene is regulated by a transcription factor protein. In case of a transcriptional repressor, the reporter protein (sfGFP) is not expressed unless a particular ligand is present and changes the conformation of the transcription factor. In case of a transcriptional activator, expression of the reporter protein is enhanced in presence of the ligand.
This regulation is a central process in nature. Because our system uses this concept, it is highly extensible and can be used to tackle various real world problems.
We consider our cell-free biosensor an outstanding instrument for the detection of harmful substances in liquids.
We see great need for a storable, transportable and easy-to-use water quality test strip all around the world.
Our development can help to improve the quality of living by revealing the quality of water.
During the reaction we had measured the fluorescence in a plate reader, in order to verify the results obtained by the app. In the nearby figures you can see: The standard CFPS setup performed not sufficent when mercury was present, HOWEVER, when using our mercury detection setup, cell-free synthesis of our reporter protein sfGFP was greatly enhanced!
Detection of Date Rape Drugs
With our developed biosensor we introduce an easy and unprecedented date rape drug ingredient biosensor! To detect the date rape drug ingredient γ-hydroxy butyrate (GHB), we had to adjust only two components of our setup: The repressor protein and the DNA sequence that the protein recognizes.
