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| {{Freiburg/CSS}} | | {{Freiburg/CSS}} |
| {{Freiburg/Menubar}} | | {{Freiburg/Menubar}} |
− | {{Freiburg/wiki_content_start}} | + | {{Team:Freiburg/wiki_content_start_bubble}} |
| <html> | | <html> |
| <style> | | <style> |
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| </style> | | </style> |
| + | |
| + | <script type="text/javascript"> |
| + | //===================BEGIN:Amazing Bubble Sidebar========================== |
| + | |
| + | $(document).ready(function(){ |
| + | // CHANGE THE FOLLOWING ATTRIBUTES // |
| + | var href_text1='https://2015.igem.org/Team:Freiburg/Results', |
| + | // Text2 needs no href as it is the actual page // |
| + | img_url='https://static.igem.org/mediawiki/2015/1/1e/Freiburg_icon_results_white_03.png', |
| + | href_text3='https://2015.igem.org/Team:Freiburg/Results/Immobilization', |
| + | // Text1 needs no text as it is a pic // |
| + | text2='Our Device'; |
| + | // Text3 needs no text as its always 'next' // |
| + | // HOLD ON CHANGING THINGS --JABBERWOCK // |
| + | |
| + | $('#bubble1').attr('href',href_text1); |
| + | $('#bubble1_img').attr('src', img_url); |
| + | $('#bubble3').attr('href',href_text3); |
| + | |
| + | $('#bubble2').text(text2); |
| + | }); |
| + | |
| + | //===================END:Amazing Bubble Sidebar========================== |
| + | </script> |
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| <!DOCTYPE html> | | <!DOCTYPE html> |
− | <div class="kommentar_stefan"> Korrigiert Stefan, Günter, Carina </div>
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| <!-- content goes in here --> | | <!-- content goes in here --> |
− | <div class="content_box results_page"> | + | <div class="content_box"> |
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− | <h1>Results of Our Self-Built iRIf Device</h1> | + | <h1 style="text-align:right">Results of Our Self-Built iRIf Device</h1> |
| <div class="level1"> | | <div class="level1"> |
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| </div> | | </div> |
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− | <h1>Detection of Antigen-Antibody binding using our selfmade device</h1> | + | <h2>Detection of Antigen-Antibody Binding Using our Selfmade Device</h2> |
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| <div class="level1"> | | <div class="level1"> |
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| </div> | | </div> |
| <p> | | <p> |
− | The camera used for the measurement was a Canon 50D. It was set to automatically acquire pictures at an interval of five seconds. After flushing the chamber with buffer, the exposure time was set to obtain apprx. 80% saturation (Fig. 1 A & B). | + | The camera used for the measurement was a Canon 50D. It was set to automatically acquire pictures at an interval of five seconds. After flushing the chamber with buffer, the exposure time was set to obtain apprx. 80% saturation (figure 1 A & B). |
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| The antibody solution was pipetted into the flow chamber without the use of a microfluidic pump. Instead, a syringe was loaded with 500 µl [5 µg/ml] anti-rabbit antibody solution (diluted in 1xPBS) and slowly released into the binding chamber of the device by gently dispensing it from the syringe. | | The antibody solution was pipetted into the flow chamber without the use of a microfluidic pump. Instead, a syringe was loaded with 500 µl [5 µg/ml] anti-rabbit antibody solution (diluted in 1xPBS) and slowly released into the binding chamber of the device by gently dispensing it from the syringe. |
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| <a class="accordion-section-title" href="#accordion-2">Spot analysis by hand: A step by step guidance</a> | | <a class="accordion-section-title" href="#accordion-2">Spot analysis by hand: A step by step guidance</a> |
| <div id="accordion-2" class="accordion-section-content" style="display:none; padding:15px;"> | | <div id="accordion-2" class="accordion-section-content" style="display:none; padding:15px;"> |
− | <p> This step by step guidance shows you how to determine endpoint intensities of the binding experiment as shown figure 1C. Have a look at our <a href="https://2015.igem.org/Team:Freiburg/Project/iRIf">iRIF</a> page to get all the background information about the detection method.</p> | + | <p> This step by step guidance shows you how to determine endpoint intensities of the binding experiment as shown figure 1C. Have a look at our <a href="https://2015.igem.org/Team:Freiburg/Project/iRIf">iRIf</a> page to get all the background information about the detection method.</p> |
| <ol> | | <ol> |
− | <li> During your iRIF experiment you take two pictures. One before the binding event and one after the binding event. Open both pictures in <a href="http://imagej.nih.gov/ij/">ImageJ</a>. </li> | + | <li> During your iRIf experiment you take two pictures. One before the binding event and one after the binding event. Open both pictures in <a href="http://imagej.nih.gov/ij/">ImageJ</a>. </li> |
| <li> Go to "Process" and click on "Image Calculator..." </li> | | <li> Go to "Process" and click on "Image Calculator..." </li> |
| <li> Set "Image1" to the image after the binding event and "Image2" to the image before the binding event. Choose "divide" as operation. Activate the "32-bit (float) result" and click "ok". A quotient picture will be calculated and opened. </li> | | <li> Set "Image1" to the image after the binding event and "Image2" to the image before the binding event. Choose "divide" as operation. Activate the "32-bit (float) result" and click "ok". A quotient picture will be calculated and opened. </li> |
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| <p> | | <p> |
− | A major problem that we encountered when building the device from scratch was to ensure that all components are at the proper distance and angle to each other. Correct alignment is crucial as a slight misplacement of a component may lead to lower signal strength, blurred images or, in the worst case, no signal at all. This can be difficult since our device does not rely on straight angles. We overcame this challenge by designing a case for the device that ensures the right placement of the components inside the device. We calculated all distances between the LED, lenses, camera and slide using the laws of geometrical optics and drew a vector graphic blueprint for our device. Next, we constructed a digital 3D model of the casing based on the vector blueprint to avoid an expensive and time-consuming trial and error process (Figure 4). | + | A major problem that we encountered when building the device from scratch was to ensure that all components are at the proper distance and angle to each other. Correct alignment is crucial as a slight misplacement of a component may lead to lower signal strength, blurred images or, in the worst case, no signal at all. This can be difficult since our device does not rely on straight angles. We overcame this challenge by designing a case for the device that ensures the right placement of the components inside the device. We calculated all distances between the LED, lenses, camera and slide using the laws of geometrical optics and drew a vector graphic blueprint for our device. Next, we constructed a digital 3D model of the casing based on the vector blueprint to avoid an expensive and time-consuming trial and error process (figure 4). |
| </p> | | </p> |
| <div class="image_box right"> | | <div class="image_box right"> |
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| <a href="https://static.igem.org/mediawiki/2015/4/4c/Freiburg_results-device_parts_vector.png" class="lightbox_trigger" title="results:device_parts_vector.png"><img alt="Freiburg iRIf device parts" src="https://static.igem.org/mediawiki/2015/4/4c/Freiburg_results-device_parts_vector.png" width="400"/></a> | | <a href="https://static.igem.org/mediawiki/2015/4/4c/Freiburg_results-device_parts_vector.png" class="lightbox_trigger" title="results:device_parts_vector.png"><img alt="Freiburg iRIf device parts" src="https://static.igem.org/mediawiki/2015/4/4c/Freiburg_results-device_parts_vector.png" width="400"/></a> |
| <div class="thumbcaption"> | | <div class="thumbcaption"> |
− | <strong>Figure 6</strong>: An image of the vector file used to order the parts. The vector file is used to excise parts of a 5 mm acrylic glass board with a laser. Black lines are cut out by the laser, gray lines/areas are engravings. Refer to the <a href="#device_download_links_anchor">download section</a> to downloaded the vector graphic files.</div> | + | <strong>Figure 6: An image of the vector file used to order the parts.</strong> The vector file is used to excise parts of a 5 mm acrylic glass board with a laser. Black lines are cut out by the laser, gray lines/areas are engravings. Refer to the <a href="#device_download_links_anchor">download section</a> to downloaded the vector graphic files.</div> |
| </div> | | </div> |
| </div> | | </div> |