Difference between revisions of "Team:Bielefeld-CeBiTec/Design"
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<div class="Subtitle"> | <div class="Subtitle"> | ||
<h2> The first steps</h2> | <h2> The first steps</h2> | ||
| − | <p> In the beginning we wanted to test, if it could really work to photograph fluorescence with an smartphone. Therefore we got an light engineering | + | <p> In the beginning we wanted to test, if it could really work to photograph fluorescence with an smartphone. Therefore we got an light engineering filter from the "Frauenkulturzentrum Bielefeld e.V." and put it in front of the camera and the flash. You can see different colors in the picture. So in our first impression we thought it's worth to try some other filters. </p> |
<figure style="margin:auto; width: 300px"> | <figure style="margin:auto; width: 300px"> | ||
| − | <a href="https://static.igem.org/mediawiki/2015/d/da/Bielefeld-CeBiTec_first_filtertests.png" data-lightbox="detection" data-title=" The first | + | <a href="https://static.igem.org/mediawiki/2015/d/da/Bielefeld-CeBiTec_first_filtertests.png" data-lightbox="detection" data-title=" The first filter test to get an rough impression if it could work to use two filters for fluorescence imaging. We used one filter in front of the flash and the same one in front of the camera. In the picture you can see purified GFP, sfGFP lysate and the lysate of a not induced sfGFP culture."><img src="https://static.igem.org/mediawiki/2015/d/da/Bielefeld-CeBiTec_first_filtertests.png" ></a> |
| − | <figcaption>The first | + | <figcaption>The first filteretest to get an rough impression if it could work to use two filters for fluorescence imaging. We used one filter in front of the flash and the same one in front of the camera. In the picture you can see purified GFP, sfGFP lysate and the lysate of a not induced sfGFP culture. |
</figcaption> | </figcaption> | ||
</figure> | </figure> | ||
| − | <p> Therefore we bought a lee color filter catalog and began to test different filter combinations. But before testing we decided to | + | <p> Therefore we bought a lee color filter catalog and began to test different filter combinations. But before testing we decided to measure the extinction and emission spectra from sfGFP as a basis to choose the most promising filter combination and to have a look if the smartphone flash really extincts sfGFP. The preselection was possible, because we had access to the light transmitting spectra of almost every filter. </p> |
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<figure style="margin:auto; width: 500px"> | <figure style="margin:auto; width: 500px"> | ||
| − | <a href="https://static.igem.org/mediawiki/2015/3/36/Bielefeld-CeBiTec_sfGFP_Spektra.png" data-lightbox="detection" data-title=" Emission and extinction | + | <a href="https://static.igem.org/mediawiki/2015/3/36/Bielefeld-CeBiTec_sfGFP_Spektra.png" data-lightbox="detection" data-title=" Emission and extinction spectra of sfGFP. The characteristic curve from the samsung galaxy S5 mini flash is also shown. The overlap of the different spectra shows that it is possible to excite sfGFP with a smartphone flash. "><img src="https://static.igem.org/mediawiki/2015/3/36/Bielefeld-CeBiTec_sfGFP_Spektra.png" ></a> |
| − | <figcaption>Emission and extinction | + | <figcaption>Emission and extinction spectra of sfGFP. The characteristic curve from the samsung galaxy S5 mini flash is also shown. The overlap of the different spectra shows that it is possible to excite sfGFP with a smartphone flash. |
</figcaption> | </figcaption> | ||
</figure> | </figure> | ||
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| − | <p> The next step was to create | + | <p> The next step was to create a dark environment. Therefore we did handicrafts to design a "black box". Now it was possible to take photos under same conditions and same background. </p> |
<figure style="margin:auto; width: 300px"> | <figure style="margin:auto; width: 300px"> | ||
| Line 60: | Line 60: | ||
</figure | </figure | ||
| − | <h2> The filter | + | <h2> The filter combinations </h2> |
| − | <p> The groundwork is done. Now we had to find the ideal | + | <p> The groundwork is done. Now we had to find the ideal filter combination. So we made a lot of photos with the preselection in every combination. After analyzing the photos with Fiji, we determined the optimal filter combination to be tokyo blue in front of the flash and twickenham green in front of the camera. </p> |
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<figure style="margin:auto; width: 400px"> | <figure style="margin:auto; width: 400px"> | ||
<a href="https://static.igem.org/mediawiki/2015/5/57/Bielefeld-CeBiTec_optimal_filtercombination.png " data-lightbox="detection" data-title="Test of different filtercombinations. On the top there is the picture without any filters. In the middle the picture was taken with fluorescence green in front of the camera and deep blue in front of the flash. The bottom photo was taken with the optimal filtercombinition. Tokyo blue in front of the flash and twickenham green in front of the camera"><img src="https://static.igem.org/mediawiki/2015/5/57/Bielefeld-CeBiTec_optimal_filtercombination.png" ></a> | <a href="https://static.igem.org/mediawiki/2015/5/57/Bielefeld-CeBiTec_optimal_filtercombination.png " data-lightbox="detection" data-title="Test of different filtercombinations. On the top there is the picture without any filters. In the middle the picture was taken with fluorescence green in front of the camera and deep blue in front of the flash. The bottom photo was taken with the optimal filtercombinition. Tokyo blue in front of the flash and twickenham green in front of the camera"><img src="https://static.igem.org/mediawiki/2015/5/57/Bielefeld-CeBiTec_optimal_filtercombination.png" ></a> | ||
| − | <figcaption> Test of different | + | <figcaption> Test of different filter combinations. On the top there is the picture without any filters. In the middle the picture was taken with fluorescence green in front of the camera and deep blue in front of the flash. The bottom photo was taken with the optimal filter combinition. Tokyo blue in front of the flash and twickenham green in front of the camera |
</figcaption> | </figcaption> | ||
</figure> | </figure> | ||
| − | + | <br> | |
| − | <p> As you can see in the picture above it | + | <p> As you can see in the picture above it is really important to choose the right filters. But is it only possible to photograph sfGFP and GFP? To find out, we tried to photograph monomeric red fluorescent protein (mRFP) lysate. </p> |
<figure style="margin:auto; width: 400px"> | <figure style="margin:auto; width: 400px"> | ||
| Line 82: | Line 82: | ||
| − | <p> So far so good. It works. But does it also works with small | + | <p> So far so good. It works. But does it also works with small amounts of volume and on paper? Therefore we took filter paper and put the lysate on the paper and took the photos. As you can see in the picture at the bottom, it is possible to photograph small amounts (5 µl) on paper up to a 1:1 dilution. </p> |
<figure style="margin:auto; width: 600px"> | <figure style="margin:auto; width: 600px"> | ||
| − | <a href="https://static.igem.org/mediawiki/2015/d/d7/Bielefeld-CeBiTec_test_on_paper.png" data-lightbox="detection" data-title="We added a small | + | <a href="https://static.igem.org/mediawiki/2015/d/d7/Bielefeld-CeBiTec_test_on_paper.png" data-lightbox="detection" data-title="We added a small amount of lysate to the scrap of paper and photographed it with the different filter combinations. The top photo is whithout any filter. The picture in the middle was taken with the filter combination for sfGFP and for the bottom picture the mRFP filters were used."><img src="https://static.igem.org/mediawiki/2015/d/d7/Bielefeld-CeBiTec_test_on_paper.png" ></a> |
| − | <figcaption> We added a small | + | <figcaption> We added a small amount of lysate to the scrap of paper and photographed it with the different filter combinations. The top photo is whithout any filter. The picture in the middle was taken with the filter combination for sfGFP and for the bottom picture the mRFP filters were used. |
</figcaption> | </figcaption> | ||
</figure> | </figure> | ||
| − | <p> Now we had to find out if we can photograph the fluorescence from the CFPS on | + | <p> Now we had to find out if we can photograph the fluorescence from the CFPS on paper. So after a CFPS run on paper, we took the scrap of paper and took a photo from it. </p> |
<figure style="margin:auto; width: 600px"> | <figure style="margin:auto; width: 600px"> | ||
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<h2> The 3D print </h2> | <h2> The 3D print </h2> | ||
| − | <p> Ok, the fluorescence detections works fine, but it's not really | + | <p> Ok, the fluorescence detections works fine, but it's not really practical to take the photo. The filter have to be in the right position direct in front of the camera respectively the flash. Also it has to be quite dark for high quality pictures. Therefore we designed a box and realized it with a 3D printer. The top can be changed specific for the smartphone you use. The test stripe can be placed on the push loading drawer and inserted into the box. So it's quite easy to use. You just have to put the smartphone on the top, insert the test strip an take the photo. </p> |
<figure style="margin:auto; width: 600px"> | <figure style="margin:auto; width: 600px"> | ||
Revision as of 19:25, 14 September 2015
Prototype Design
The first steps
In the beginning we wanted to test, if it could really work to photograph fluorescence with an smartphone. Therefore we got an light engineering filter from the "Frauenkulturzentrum Bielefeld e.V." and put it in front of the camera and the flash. You can see different colors in the picture. So in our first impression we thought it's worth to try some other filters.
Therefore we bought a lee color filter catalog and began to test different filter combinations. But before testing we decided to measure the extinction and emission spectra from sfGFP as a basis to choose the most promising filter combination and to have a look if the smartphone flash really extincts sfGFP. The preselection was possible, because we had access to the light transmitting spectra of almost every filter.
The next step was to create a dark environment. Therefore we did handicrafts to design a "black box". Now it was possible to take photos under same conditions and same background.
The groundwork is done. Now we had to find the ideal filter combination. So we made a lot of photos with the preselection in every combination. After analyzing the photos with Fiji, we determined the optimal filter combination to be tokyo blue in front of the flash and twickenham green in front of the camera.
As you can see in the picture above it is really important to choose the right filters. But is it only possible to photograph sfGFP and GFP? To find out, we tried to photograph monomeric red fluorescent protein (mRFP) lysate.
Does it work on Paper?
So far so good. It works. But does it also works with small amounts of volume and on paper? Therefore we took filter paper and put the lysate on the paper and took the photos. As you can see in the picture at the bottom, it is possible to photograph small amounts (5 µl) on paper up to a 1:1 dilution.
Now we had to find out if we can photograph the fluorescence from the CFPS on paper. So after a CFPS run on paper, we took the scrap of paper and took a photo from it.
The 3D print
Ok, the fluorescence detections works fine, but it's not really practical to take the photo. The filter have to be in the right position direct in front of the camera respectively the flash. Also it has to be quite dark for high quality pictures. Therefore we designed a box and realized it with a 3D printer. The top can be changed specific for the smartphone you use. The test stripe can be placed on the push loading drawer and inserted into the box. So it's quite easy to use. You just have to put the smartphone on the top, insert the test strip an take the photo.
