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Team:Harvard/Hardware/MFCProcedure
From 2008.igem.org
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'''Procedure''' | '''Procedure''' | ||
| - | + | 1) Mill Polycarbonate | |
| - | + | * Cut polycarbonate sheet into 4 equal 3" x 3" pieces | |
| - | + | * Drill four 3/8" holes through each piece, 1 per corner, indented 5mm from both sides | |
| + | * Drill a 1/2" hole in the center of each piece | ||
[[Image:endplates.jpg | 600px]] | [[Image:endplates.jpg | 600px]] | ||
| - | + | * Cut polycarbonate tube into two equal 2" halves | |
| - | + | * Drill four 1/4" holes through each half in configuration shown | |
[[Image:drilled_tube.jpg | 600px]] | [[Image:drilled_tube.jpg | 600px]] | ||
| - | + | * Tap each hole with 1/4" -28 spiral tap | |
| - | + | 2) Glue Chambers (repeat for each half) | |
| - | + | * Center tube on endplate by marking plate with 'X' from corner to corner | |
| - | + | * Squirt 2mm thick line of silicone on edge of tube (edge furthest from holes) | |
| - | + | * Press tube firmly against marked location on endplate | |
| - | + | * Quickly spread excess silicone along edge | |
| - | + | * Let stand 24h to harden | |
[[Image:glued_half.jpg | 600px]] | [[Image:glued_half.jpg | 600px]] | ||
| - | + | 3) Construct Gaskets | |
| - | + | * Cut silicone sheet into two equal 2.25" x 2.25" pieces | |
| - | + | * Cut out centered inner squares in each piece, 1.75" x 1.75" | |
| - | + | * Using inner squares, cut two 'O' rings, inner diameter 1/4", outer diameter 1/2" | |
[[Image:gaskets.jpg | 600px]] | [[Image:gaskets.jpg | 600px]] | ||
| - | + | 4) Construct Electrodes | |
| - | + | * Cut titanium wire into one 8" piece and one 16" piece | |
| - | + | * Using pliers, shape anode and cathode as shown | |
'''Anode Frame'''<br> | '''Anode Frame'''<br> | ||
[[Image:Anodeframe.jpg | 500px]]<br> | [[Image:Anodeframe.jpg | 500px]]<br> | ||
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[[Image:Cathodeframe.jpg | 500px]] | [[Image:Cathodeframe.jpg | 500px]] | ||
| - | + | * Spear carbon felt with tip of anode titanium wire and wedge into frame | |
| - | + | * Weave platinum carbon cloth through cathode titanium wire | |
'''Anode'''<br> | '''Anode'''<br> | ||
[[Image:Anode_electrode.jpg | 500px]]<br> | [[Image:Anode_electrode.jpg | 500px]]<br> | ||
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[[Image:Cathode_electrode.jpg | 500px]] | [[Image:Cathode_electrode.jpg | 500px]] | ||
| - | + | 5) Seal Injection Ports | |
| - | + | * Wrap threads of all eight Luer Lock screws with 1' of teflon tape in opposite direction of screwing | |
| - | + | * Screw Luer Locks into all tapped holes in both chambers | |
[[Image:fin_chamber.jpg | 600px]] | [[Image:fin_chamber.jpg | 600px]] | ||
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===Assembling Chambers=== | ===Assembling Chambers=== | ||
| - | '''Procedure''' | + | '''Procedure'''<br> |
1) Prepare Electrodes | 1) Prepare Electrodes | ||
* Attach Luer Lock injection ports to all chamber screws | * Attach Luer Lock injection ports to all chamber screws | ||
* Poke tip of electrodes through designated ports from the inside | * Poke tip of electrodes through designated ports from the inside | ||
[[Image:chamber_w_elec.jpg | 600px]] | [[Image:chamber_w_elec.jpg | 600px]] | ||
| - | + | <br> | |
| - | 2 Align Chambers | + | 2) Align Chambers |
* Lay one chamber on a flat surface | * Lay one chamber on a flat surface | ||
* Place silicone square ring on top edge of tube | * Place silicone square ring on top edge of tube | ||
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* Center second silicone square ring on polycarbonate square | * Center second silicone square ring on polycarbonate square | ||
* Set second chamber on top of silicone, ensuring ports facing same direction as first chamber | * Set second chamber on top of silicone, ensuring ports facing same direction as first chamber | ||
| - | + | <br> | |
3) Clamp Chambers | 3) Clamp Chambers | ||
* Move assembly into vice or clamp | * Move assembly into vice or clamp | ||
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#* 15.1185 g/L 50mM PIPES (hydrogen) | #* 15.1185 g/L 50mM PIPES (hydrogen) | ||
#* ''7.0 pH'' | #* ''7.0 pH'' | ||
| - | |||
# Phosphate buffer (60ml / fuel cell) | # Phosphate buffer (60ml / fuel cell) | ||
#* 2.918 g/L Monosodium phosphate, monohydrate | #* 2.918 g/L Monosodium phosphate, monohydrate | ||
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#* 5.844 g/L 100mM NaCl | #* 5.844 g/L 100mM NaCl | ||
#* ''7.0 pH'' | #* ''7.0 pH'' | ||
| + | # Resazurin solution | ||
| + | #* 0.489 g/L 0.8mM Resazurin | ||
| + | # Lactate Solution | ||
| + | #* 13.51 g/L Lactic acid | ||
| + | #* neutralize to pH 7 with NaOH | ||
===Gas Tubing Assembly=== | ===Gas Tubing Assembly=== | ||
| - | Materials | + | '''Materials''' |
* 25' Silicone Soft Rubber Tubing, 3/32" ID, 7/32" OD, 1/16" Wall | * 25' Silicone Soft Rubber Tubing, 3/32" ID, 7/32" OD, 1/16" Wall | ||
* Tank of Compressed Nitrogen | * Tank of Compressed Nitrogen | ||
| Line 212: | Line 217: | ||
* 2 Rubber Stoppers | * 2 Rubber Stoppers | ||
| - | Procedure | + | '''Procedure''' |
| - | + | 1) Make Flow Regulators | |
| - | + | * Insert nozzle of female Luer Lock into rubber stoppers (poke hole if necessary) | |
| - | + | * Cap aspirator flasks with rubber stoppers | |
| - | + | * Attach tubing from gas sources to each glass nozzle of aspirator flask | |
[[Image:flow_regs.jpg | 600px]] | [[Image:flow_regs.jpg | 600px]] | ||
| - | + | 2) Make Manifolds | |
| - | + | * Attach T-junction Luer Lock pieces into manifold (1 junction/ fuel cell ; 2 manifods total) | |
| - | + | * Turn last juction such that off is facing end of manifold | |
| - | + | * Attach tubing from stopper of flow regulators to beginning of each mainfold | |
[[Image:manifolds.jpg | 600px]] | [[Image:manifolds.jpg | 600px]] | ||
Latest revision as of 04:10, 30 October 2008
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