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| {{Freiburg/Menubar}} | | {{Freiburg/Menubar}} |
− | {{Freiburg/wiki_content_start}} | + | {{Team:Freiburg/wiki_content_start_bubble}} |
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| </style> | | </style> |
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| + | <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> |
| <!-- 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 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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| <p> | | <p> |
− | Here you will see what we were able to measure with the very first prototype of our self-built device. To see the results of our final device, please refer to the <a href="https://2015.igem.org/wiki/index.php?title=Team:Freiburg/Results#device_results_anchor">essential results page</a>. A detailed explanation on how to build your own low-priced iRIf device can be found on the <a href="#howto_anchor">bottom of this page</a>.
| + | The observation and quantification of protein-protein or other molecular interactions is a very difficult task and requires expensive and complicated equipment. Therefore, we designed and <a href="#howto_anchor">constructed a cheap and easy to use device </a> that can easily be reconstructed by the iGEM community. Using our device we were able to measure <a href="https://2015.igem.org/wiki/index.php?title=Team:Freiburg/Results#device_results_anchor">antigen-antibody interactions</a> and reproduced an experiment performed with the professional device. All in all, we made it easy for everyone to perform professional binding experiments. |
| </p> | | </p> |
| </div> | | </div> |
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− | <h1>Our First Prototype is Able to Detect Antigen-Antibody Binding</h1> | + | <h2>Detection of Antigen-Antibody Binding Using our Selfmade Device</h2> |
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| <div class="level1"> | | <div class="level1"> |
| <div class="image_box left"> | | <div class="image_box left"> |
− | <div class="thumb2 trien" style="width:400px"><div class="thumbinner"><a href="https://static.igem.org/mediawiki/2015/f/fc/Freiburg_results-result_device_20150818.jpg" title="results:result_device_20150818.jpg" class="media lightbox_trigger"><img alt="Our device measurement with rabbit proteins" src="https://static.igem.org/mediawiki/2015/f/fc/Freiburg_results-result_device_20150818.jpg" width="400"/></a><div class="thumbcaption"><strong>Figure 1: Binding of anti-rabbit antibodies to rabbit proteins measured in our device.</strong> A: The first picture of the measurement. B: The last picture of the measurement. C: The quotient picture of the first and last picture. D: Schematic illustration of the pattern of the spots on the slide </div></div></div> | + | <div class="thumb2 trien" style="width:400px"><div class="thumbinner"><a href="https://static.igem.org/mediawiki/2015/f/fc/Freiburg_results-result_device_20150818.jpg" title="results:result_device_20150818.jpg" class="media lightbox_trigger"><img alt="Our device measurement with rabbit proteins" src="https://static.igem.org/mediawiki/2015/c/cc/Freiburg_results-result_device_20150818_preview.jpg" width="400"/></a><div class="thumbcaption"><strong>Figure 1: Binding of anti-rabbit antibodies to rabbit proteins measured in our device.</strong> A: The first picture of the measurement. B: The last picture of the measurement. C: The quotient picture of the first and last picture. D: Schematic illustration of the spot pattern on the slide </div></div></div> |
| </div> | | </div> |
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| <p> | | <p> |
− | For testing our device we immobilized proteins derived from rabbits on an iRIf slide in distinct spots (figure 1 D). The proteins we used were polyclonal antibodies against HCV (Hepatitis C Virus) derived from rabbits. We aimed to detect these proteins with anti-rabbit antibodies (antibodies which specifically bind the constant region of antibodies dervied from rabbits). We used these interaction partners in this series of experiments to reproduce a successful measurement, which we previously performed in the professional measuring device. | + | In order to test our device, we immobilized rabbit-derived proteins on an iRIf slide in distinct spots (figure 1 D). These proteins were polyclonal antibodies against HCV (Hepatitis C Virus) raised in rabbit. They were detected using anti-rabbit antibodies that bind specifically to the constant regions of all rabbit derived antibodies. We used them as interaction partners in a series of experiments to reproduce a measurement previously performed in the professional device. |
− | The binding layer on the iRIf slide consisted of an APTES/PDITC surface. The spots on the slide were produced by pipetting 3 µl [500 µg/ml] rabbit-anti-HCV protein and 3 µl [1 mg/ml] BSA in an alternating pattern onto the slide. After incubation o/n the slide was blocked for 30 min in BSA [10 mg/ml] solution. | + | The binding layer on the iRIf slide consisted of an APTES/PDITC surface. The spots on the slide were produced by pipetting 3 µl [500 µg/ml] rabbit anti-HCV protein and 3 µl [1 mg/ml] BSA onto the slide in an alternating pattern. After incubation o/n the slide was blocked for 30 min with BSA [10 mg/ml]. |
| </p> | | </p> |
| <div class="image_box right" style="clear:both;"> | | <div class="image_box right" style="clear:both;"> |
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| </div> | | </div> |
| <p> | | <p> |
− | The camera used for the measurement was a Canon 50D. The camera was set to automatically take pictures at an interval of five seconds. The exposure time was set in order for the pixels in the image to have approx. 80% of maximum light saturation before the solution was flushed onto the chip (figure 1 C & D). | + | 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 flowchamber without the use of any microfluidic pump. Instead, a syringe was loaded with 500 µl [5 µg/ml] anti-rabbit antibody solution (diluted in PBS) 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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− | As can be seen in figure 1 C, the quotient picture clearly shows binding of anti-rabbit antibodies to the rabbit protein spots. The BSA control spots show none or negligible unspecific binding. | + | As can be seen in figure 1 C, the quotient picture clearly shows binding of anti-rabbit antibodies to the rabbit protein spots. The BSA control spots show no or negligible unspecific binding. |
| </p> | | </p> |
| + | |
| + | <div class="accordion" style="margin-top:100px"> |
| + | <div class="accordion-section"> |
| + | <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;"> |
| + | <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> |
| + | <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> 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> Go to "Image" then "Adjust" and click on "Brightness/Contrast..." and play around with "brightness" and "contrast" till the spots get visible.</li> |
| + | <li> Mark the spots using the "oval" selection tool and press "T" on your keyboard. Repeat this for every spot. Do not forget to make a selection of the background. </li> |
| + | <li> Go to the ROI-manager that popped up when you first pushed T and click on "Deselect" and then on "Measure". The results of your measurement will pop up. You can save them in Excel.</li> |
| + | </ol> |
| + | <p> |
| + | Remember that signals normally differ by about 0.1 - 0.8 percent depending on how strong your binding is. |
| + | </p> |
| + | |
| + | </div><!-- end accordion-section-content --> |
| + | </div><!-- end accordion-section --> |
| + | </div> <!-- end accordion --> |
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| <span class="anchor" id="howto_anchor"></span> | | <span class="anchor" id="howto_anchor"></span> |
− | <h1 class="left" id="how_to_build_your_own_device">How To Build Your Own Device</h1> | + | <h1 class="left" id="how_to_build_your_own_device">How To Build Your Own iRIf Device</h1> |
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| <p> | | <p> |
− | In this section we will demonstrate how to build your own iRIf device from scratch, using affordable, low-tech material. The design is focused on creating a device that is both low-priced and portable, demonstrating the potential for the DiaCHIP to be used even in rural areas where high-tech laboratories are poorly accessible. | + | In this section we demonstrate how to build your own iRIf device from scratch, using affordable, low-tech material. The design is focused on creating a device that is both low-priced and portable. Therefore the DiaCHIP can even be used in areas where high-tech laboratories are inaccessible. |
| </p> | | </p> |
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| </a> | | </a> |
| <div class="thumbcaption"> | | <div class="thumbcaption"> |
− | <strong>Figure 3: An illustration showing the exact setup of our device from a top perspective.</strong> | + | <strong>Figure 3: Our device from top.</strong> |
| </div> | | </div> |
| </div> | | </div> |
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| <p> | | <p> |
− | As can be seen in figure 3, the basic setup is fairly simple. Light from an LED enters a lense to obtain a parallel light beam. To achieve this, the distance from the LED to the lense has to be exactly one focal length. | + | As seen in figure 3, the basic setup is fairly simple. Light from an LED enters a lens to obtain a parallel light beam. To achieve this, the distance from the LED to the lens should be close to one focal length. |
− | The light hits the iRIf slide afterwards where it is reflected (in the same angle as it hits the slide) and enters a second lense, whose purpose is to project a sharp image of the slide onto the CCD chip of the camera. | + | Next, the light hits the iRIf slide where it is reflected (under the same angle as it hits the slide) and enters a second lens that projects an image of the slide onto the CCD chip of the camera. |
| </p> | | </p> |
| </div> | | </div> |
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| <table class="inline"> | | <table class="inline"> |
| <tr class="row0"> | | <tr class="row0"> |
− | <th class="col0"> Part </th><th class="col1"> Description </th><th class="col2"> Manufacturer / Partname </th><th class="col3"> Price[USD]</th> | + | <th class="col0"> Part </th><th class="col1"> Description </th><th class="col2"> Manufacturer / Part name </th><th class="col3"> Price[USD]</th> |
| </tr> | | </tr> |
| <tr class="row1"> | | <tr class="row1"> |
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| </tr> | | </tr> |
| <tr class="row7"> | | <tr class="row7"> |
− | <td class="col0">A microfluidics chamber</td><td class="col1">Our chamber consists of an iRIf slide attached to a PDMS flowcell</td><td class="col2">Made ourselves</td><td class="col3"></td> | + | <td class="col0">A microfluidics chamber</td><td class="col1">Our chamber consists of an iRIf slide attached to a PDMS flow cell</td><td class="col2">Made ourselves</td><td class="col3"></td> |
| </tr> | | </tr> |
| <tr class="row8"> | | <tr class="row8"> |
− | <td class="col0">Magnets</td><td class="col1">Used to attach the flowcell to the device</td><td class="col2">5 mm Neodymium magnets</td><td class="col3">3</td> | + | <td class="col0">Magnets</td><td class="col1">Used to attach the flow cell to the device</td><td class="col2">5 mm Neodymium magnets</td><td class="col3">3</td> |
| </tr> | | </tr> |
| <tr class="row9"> | | <tr class="row9"> |
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| </tr> | | </tr> |
| <tr class="row10"> | | <tr class="row10"> |
− | <td class="col0">A microfluidic tube</td><td class="col1"> Used to connect the flowchamber with the syringe</td><td class="col2">-</td><td class="col3">< 1</td> | + | <td class="col0">A microfluidic tube</td><td class="col1"> Used to connect the flow chamber with the syringe</td><td class="col2">-</td><td class="col3">< 1</td> |
| </tr> | | </tr> |
| <tr class="row11"> | | <tr class="row11"> |
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| <div class="thumb2 trien" style="width:610px"><div class="thumbinner"><a class="media lightbox_trigger" href="https://static.igem.org/mediawiki/2015/2/24/Freiburg_results-led_and_magnets_and_flowcell.png" title="results:led_and_magnets_and_flowcell.png"><img alt="Some parts needed to build your own iRIf device" class="mediabox2" src="https://static.igem.org/mediawiki/2015/2/24/Freiburg_results-led_and_magnets_and_flowcell.png" width="600"/></a> | | <div class="thumb2 trien" style="width:610px"><div class="thumbinner"><a class="media lightbox_trigger" href="https://static.igem.org/mediawiki/2015/2/24/Freiburg_results-led_and_magnets_and_flowcell.png" title="results:led_and_magnets_and_flowcell.png"><img alt="Some parts needed to build your own iRIf device" class="mediabox2" src="https://static.igem.org/mediawiki/2015/2/24/Freiburg_results-led_and_magnets_and_flowcell.png" width="600"/></a> |
| <div class="thumbcaption"> | | <div class="thumbcaption"> |
− | <strong>Figure 3:</strong> (A) The LED glued to the cooling element with thermal adhesive; (B) The 10 magnets, each 5x5x5 mm as well as a PDMS flowchamber (depth of the flowchamber: 30 µm); (C) One of the lenses used in the setup</div></div></div> | + | <strong>Figure 3:</strong> (A) The LED glued to the cooling element with thermal adhesive; (B) The 10 magnets, each 5x5x5 mm as well as a PDMS flow chamber (depth of the flow chamber: 30 µm); (C) One of the lenses used in the setup</div></div></div> |
| </div> | | </div> |
| </div><!-- end accordion-section-content --> | | </div><!-- end accordion-section-content --> |
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| <p> | | <p> |
− | A major problem that we confronted when building the device from scratch was to assure that all components are at the exact distance and angle to each other. This 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 problem by designing a case for the device that ensures the right placement of the components inside the device. This was achieved by calculating all the distances beforehand using physical laws of optics. We realized this by drawing an exact vector graphic blueprint for our device. We then constructed a digital 3D model of the casing based on the vector blueprint to avoid a costly 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"> |
| <div class="thumb2 trien" style="width:410px"><div class="thumbinner"><a class="media lightbox_trigger" href="https://static.igem.org/mediawiki/2015/6/69/Freiburg_results-device_3d_perp.png" title="results:device_3d_perp.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/6/69/Freiburg_results-device_3d_perp.png" width="400"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/6/69/Freiburg_results-device_3d_perp.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div><strong>Figure 4:</strong> 3D model of our device, built from the vector files used to order the parts.</div></div></div> | | <div class="thumb2 trien" style="width:410px"><div class="thumbinner"><a class="media lightbox_trigger" href="https://static.igem.org/mediawiki/2015/6/69/Freiburg_results-device_3d_perp.png" title="results:device_3d_perp.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/6/69/Freiburg_results-device_3d_perp.png" width="400"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/6/69/Freiburg_results-device_3d_perp.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div><strong>Figure 4:</strong> 3D model of our device, built from the vector files used to order the parts.</div></div></div> |
− | | + | </div> |
− | | + | <div class="image_box right"> |
− | <div class="thumb2 trien" style="width:410px;padding-top:10px"><div class="thumbinner"><a class="media lightbox_trigger" href="https://static.igem.org/mediawiki/2015/8/88/Freiburg_results-device_3d_no_walls.png" title="results:device_3d_no_walls.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/8/88/Freiburg_results-device_3d_no_walls.png" width="400"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/8/88/Freiburg_results-device_3d_no_walls.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div><strong>Figure 5: The 3D model of our device without walls and top part.</strong> Parts have been colorized for clarification - Green: The wall where light exits the device and enters the camera; Pink: a platform holding Cooling-Element+LED in place; Blue: platforms for holding the lenses in place; Gray: rear end wall where the flowchamber is attached to. The slits in the top and bottom part where the magnets have to be fixed</div></div></div> | + | <div class="thumb2 trien" style="width:410px;padding-top:10px"><div class="thumbinner"><a class="media lightbox_trigger" href="https://static.igem.org/mediawiki/2015/8/88/Freiburg_results-device_3d_no_walls.png" title="results:device_3d_no_walls.png"><img alt="" class="mediabox2" src="https://static.igem.org/mediawiki/2015/8/88/Freiburg_results-device_3d_no_walls.png" width="400"/></a><div class="thumbcaption"><div class="magnify"><a class="internal" href="https://static.igem.org/mediawiki/2015/8/88/Freiburg_results-device_3d_no_walls.png" title="vergrößern"><img alt="" height="11" src="/igem2015/lib/plugins/imagebox/magnify-clip.png" width="15"/></a></div><strong>Figure 5: The 3D model of our device without walls and top part.</strong> Parts have been colorized for clarification - Green: The wall where light exits the device and enters the camera; Pink: a platform holding the cooling element and LED in place; Blue: platforms for holding the lenses in place; Gray: rear end wall where the flow chamber is attached to. The slits in the top and bottom part where the magnets need to be attached.</div></div></div> |
| </div> | | </div> |
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| <p> | | <p> |
− | We designed the casing so that all the necessary parts (lenses, LED) are held in the correct position safely during the measurement, but remain removable to grant easy transportation of the device (i.e. to the Giant Jamboree). Figure 5 illustrates the parts that hold the lenses and LED in place. | + | The casing is designed to keep all the necessary parts (lenses, LED) safely in place during a measurement, but allows them to be removed to facilitate transportation of the device (i.e. to the Giant Jamboree). Figure 5 illustrates the parts that hold the lenses and LED in place. |
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| </p> | | </p> |
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| <p> | | <p> |
− | After assuring that the 3D model of our device was set-up correctly, we created a vector graphic file which layed out all the parts needed for the casing in a 2D plain. Using this vector graphic file, we ordered the parts at <a href="http://www.formulor.com" target="_blank">Formulor</a>, a service which uses laser cutting to cut out parts from acrylic glass. The cutting laser is guided by the vector paths defined in the vector graphic template. The vector graphic template is shown in figure 6, and may be <a href="#device_download_links_anchor">downloaded</a> and used by everyone to build their own device. To allow easy mounting of the casing we also provide an easy to understand <a href="https://static.igem.org/mediawiki/2015/2/2f/Freiburg_iRIf_Device_Manual.pdf" target="_blank">manual</a>. One advantage of using acrylic glass is that the parts can be glued together easily using a few drops of acetone, fusing the parts together. The vector graphic file used to laser our the casing parts also contains a template for the parts which are necessary to attach the glass and PDMS slide to the device. These parts have to be cut from 1 mm thick acrylic glass. Once they are cut from the acrylic glass with a laser, the microfluidic tubes can be glued to the parts (figure 7). Once this is completed, these parts can be attached to the device with the magnets (figure 8)</p> | + | Subsequently to the setup of the 3D model of our device, we created a vector graphic of all required parts in a 2D plane. Using this vector graphic, we ordered the parts at <a href="http://www.formulor.com" target="_blank">Formulor</a>, a service which uses laser cutting to cut out parts from acrylic glass and other materials. The cutting laser is guided by the paths defined in the vector graphic template. The template is shown in figure 6, and may be <a href="#device_download_links_anchor">downloaded</a> and used by everyone for building their own device. To allow easy mounting of the casing, we also provide an easy to understand <a href="https://static.igem.org/mediawiki/2015/2/2f/Freiburg_iRIf_Device_Manual.pdf" target="_blank">manual</a>. One advantage of acrylic glass is that the parts can be glued together easily using a few drops of acetone. The vector graphic also contains a template for the parts which are necessary to attach the glass and PDMS slide to the device. They should be cut from 1 mm thick acrylic glass. The microfluidic tubes can be glued to these parts (figure 7) and attached to the device with the magnets (figure 8)</p> |
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| <div class="image_box left"> | | <div class="image_box left"> |
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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> |
| </div> | | </div> |
| <p> | | <p> |
− | The most difficult part to build by oneself is the PDMS flowchamber. For producing such flowchambers, a wafer has to be made in a cleanroom. The template used for our PDMS flowcell is shown in figure 9. If your facility has an engineering department, there is a high chance that a cleanroom is present. Ask the personel in charge if they can help you out with producing your flowcell. If you already use another type of microfluidic flowchambers, our device should still be compatible with it, though some adaptions might be necessary. If you have no possibility of creating a flowchamber, you might want to consider building a very simplified DIY flowchambers from two glass slides, separated by thin duct tape. Note that our device was not primarily build to support such a chamber though and would need appropriate modifications. | + | The most difficult part to build is the PDMS flow chamber since it requires a microstructured silicon wafer, which can only be produced in a cleanroom environment. The template used for our PDMS flow cell is shown in figure 9. If your facility has an engineering department, there is a high chance that a cleanroom is present. Ask the personnel in charge if they can help you out with producing your flow cell. If you already use another type of microfluidic flow chamber, our device should still be compatible with it, though some adaptions might be necessary. If you have no possibility of creating a flow chamber, you might want to consider building a very simplified DIY flow chambers from two glass slides, separated by thin tape. Note that our device was not primarily build to support such a chamber and would need appropriate modifications. |
| </p> | | </p> |
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| <a href="https://static.igem.org/mediawiki/2015/2/27/Freiburg_Device_Flowcell_chamber_syringe.jpg" class="lightbox_trigger" title="Flow cell attachment"><img alt="Freiburg iRIf device flow cell attachment" src="https://static.igem.org/mediawiki/2015/0/08/Freiburg_Device_Flowcell_chamber_syringe_preview.jpg" width="400"/></a> | | <a href="https://static.igem.org/mediawiki/2015/2/27/Freiburg_Device_Flowcell_chamber_syringe.jpg" class="lightbox_trigger" title="Flow cell attachment"><img alt="Freiburg iRIf device flow cell attachment" src="https://static.igem.org/mediawiki/2015/0/08/Freiburg_Device_Flowcell_chamber_syringe_preview.jpg" width="400"/></a> |
| <div class="thumbcaption"> | | <div class="thumbcaption"> |
− | <strong>Figure 7</strong> - left: a flow chamber consisting of an iRIf slide attached to a PDMS flowcell - center: a syringe attached to a pipette tip, used to inject antibody solutions into the flowchamber - right: the part of our device which connects the flowchamber, the syringe and the iRIf device. | + | <strong>Figure 7</strong> - left: a flow chamber consisting of an iRIf slide attached to a PDMS (red) flow cell - center: a syringe attached to a pipette tip, used to inject antibody solutions into the flow chamber - right: the part of our device which connects the flow chamber, the syringe and the iRIf device. |
| </div> | | </div> |
| </div> | | </div> |
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| <a href="https://static.igem.org/mediawiki/2015/c/c0/Freiburg_flowchamber_mounted_unmounted.png" class="lightbox_trigger" title="Mounted and unmounted flow-chamber in our iRIf device"><img alt="Freiburg iRIf device flow chamber mounted and unmounted" src="https://static.igem.org/mediawiki/2015/1/1f/Freiburg_flowchamber_mounted_unmounted_preview.png" width="390"/></a> | | <a href="https://static.igem.org/mediawiki/2015/c/c0/Freiburg_flowchamber_mounted_unmounted.png" class="lightbox_trigger" title="Mounted and unmounted flow-chamber in our iRIf device"><img alt="Freiburg iRIf device flow chamber mounted and unmounted" src="https://static.igem.org/mediawiki/2015/1/1f/Freiburg_flowchamber_mounted_unmounted_preview.png" width="390"/></a> |
| <div class="thumbcaption"> | | <div class="thumbcaption"> |
− | <strong>Figure 8:</strong> The device with and without the mounted flowchamber. | + | <strong>Figure 8:</strong> The device with and without the mounted flow chamber. |
| </div> | | </div> |
| </div> | | </div> |
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| <a href="https://static.igem.org/mediawiki/2015/0/00/Freiburg_Masken_M25-M33-1.jpg" class="lightbox_trigger" title="PDMS flow chamber"><img alt="Freiburg iRIf device flow cell" src="https://static.igem.org/mediawiki/2015/0/00/Freiburg_Masken_M25-M33-1.jpg" width="400"/></a> | | <a href="https://static.igem.org/mediawiki/2015/0/00/Freiburg_Masken_M25-M33-1.jpg" class="lightbox_trigger" title="PDMS flow chamber"><img alt="Freiburg iRIf device flow cell" src="https://static.igem.org/mediawiki/2015/0/00/Freiburg_Masken_M25-M33-1.jpg" width="400"/></a> |
| <div class="thumbcaption"> | | <div class="thumbcaption"> |
− | <strong>Figure 9:</strong> Layout used to produce the wafer of our PDMS flowcell. | + | <strong>Figure 9:</strong> Layout used to produce the wafer of our PDMS flow cell. |
| </div> | | </div> |
| </div> | | </div> |
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| </div> | | </div> |
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− | <h2>Legal notice</h2> | + | <h2 class="left">Legal notice</h2> |
| <p> | | <p> |
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