Team:EPF-Lausanne/Tools/Microfluidics/HowTo1

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== PDMS chip fabrication ==
== PDMS chip fabrication ==
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If your lab is equipped, you should have no problem replicating our [[Team:EPF-Lausanne/Protocols/PDMS_two_layer_device_fabrication|protocol]] for MITOMI chip fabrication, illustrated here by Lilia:
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== PDMS chip fabrication ==
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MITOMI chips are two layer devices. A thick upper layer is imprinted with the flow channels (those that will contain reagents) and a thin bottom layer is imprinted with the control channels (used to actuate on-chip valves). The role of each layer is explained further in [[Team:EPF-Lausanne/Tools/Microfluidics/HowTo2|Part II]].
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<embed type="application/x-shockwave-flash" src="https://picasaweb.google.com/s/c/bin/slideshow.swf" width="600" height="400" flashvars="host=picasaweb.google.com&captions=1&hl=en_US&feat=flashalbum&RGB=0x000000&feed=https%3A%2F%2Fpicasaweb.google.com%2Fdata%2Ffeed%2Fapi%2Fuser%2F114670117111403230486%2Falbumid%2F5654596587940840561%3Falt%3Drss%26kind%3Dphoto%26hl%3Den_US" pluginspage="http://www.macromedia.com/go/getflashplayer"></embed>
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<p> 1)    Place molds into a TMCS vapor chamber for at least 20min</p>
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Casting PDMS involves mixing a precursor and a curing agent (sold as a kit from XXX), degassing, pouring into the mould (or spin-coating onto the wafer for the thin layers), and partial curing at 80°C. After curing, holes are punched through the flow layer, to allow plugging in of external tubing (again, see part II). The two layers are then superposed and aligned by hand, so that the valves from the control layer overlap the correct channels on the flow layer.
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<p> 2a) Control layer mixture</p>
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<p> 2b)  Mix for 1 minute, degas for 2 minutes (standard protocol)</p>
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<p> 2c)  Pour onto control layer mold and place mold in vacuum chamber for at least 20min</p>
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<p> 3a) Flow layer mixture </p>
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<p> 3b)  Mix for 1 minute and degas for 2 minutes (standard protocol)</p>
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<p> 3c)  Spin coat onto flow layer</p>
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<p> 4)    Remove control layer mold from vacuum chamber, making sure no bubbles are left on the surface (remove with a toothpick if you see some )</p>
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<p> 5)    Place the control and flow layer in a 80C convection oven and incubate for 30 minutes (timing is critical here!)</p>
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<p> 6a) Remover wafers both layers from the oven, cut out control layer </p>
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<p> 6b)  Punch holes </p>
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<p> 6c)  Align control to the flow layer</p>
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<p> 7)    Put aligned device back into 80C oven and incubate for at least 90 minutes (and here you can increase the baking time)</p>
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<p> 8a)  Remove devices from oven</p>
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<p> 8b)  Cut them out </p>
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<p> 8c)  Punch flow layer holes and cut the edges through the flow layer</p>
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After alignment, curing is finalised. This fully solidifies the chip and ensures a strong bond between the two layers. After curing, holes are punched through to the flow layer. For most chips, the last step is to bond them them to a glass microscope slide, following a plasma surface treatment if strong bonding is required. In the case of the MITOMI chips, they are instead aligned onto a spotted array from a DNA library. At this point, the chip is ready to use.
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Before use the chip should be aligned to the epoxy-treated glass slide. There can be DNA, bacteria or other substances spotted on it ( in this project for the MITOMI experiences we spot different variants of the tetO sequence, we use QArray robot to do it).
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The specifics of each step are explained in our [[Team:EPF-Lausanne/Protocols/PDMS_two_layer_device_fabrication|protocol]] for MITOMI chip fabrication. The process is illustrated on our [https://picasaweb.google.com/114670117111403230486/ChipFab_steps?authuser=0&feat=directlink Picasa gallery]. More pictures of chip fabrication are also available [https://picasaweb.google.com/114670117111403230486/ChipFabMITOMI?authuser=0&feat=directlink here].
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<embed type="application/x-shockwave-flash" src="https://picasaweb.google.com/s/c/bin/slideshow.swf" width="600" height="400" flashvars="host=picasaweb.google.com&captions=1&hl=en_US&feat=flashalbum&RGB=0x000000&feed=https%3A%2F%2Fpicasaweb.google.com%2Fdata%2Ffeed%2Fapi%2Fuser%2F114670117111403230486%2Falbumid%2F5654596587940840561%3Falt%3Drss%26kind%3Dphoto%26hl%3Den_US" pluginspage="http://www.macromedia.com/go/getflashplayer"></embed>
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Revision as of 00:30, 22 September 2011