Materials

• Laser-cut acrylic arena pieces
• Base, frame, and top



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The top piece may differ depending on the type of arena being assembled
• Methylethylketone (MEK), located on the first shelf above lab bench 2
• Gloves
• p100 micropipette with tips
• Kimwipes
• 70% ethanol
• Lab tape
• Sharpie

Protocol

1. Carefully remove the brown layer from each side of each arena piece.
2. Set the p100 to 50 µL.
3. While wearing gloves, line up the wall piece with the engraved side on the base piece.
4. To distinguish the engraved side from the smooth side, you can try to feel the texture with your hand. If that is too hard to distinguish, hold the piece up to a light parallel to the countertop and observe which side has the engraved white line on the top instead of the bottom.

6. Fill the pipette tip with MEK.
7. While firmly holding the frame to the base, slide the pipette tip along one of the edges while releasing MEK into the grooves between the layers
• You should be able to see a small color change where the MEK is embedded into the grooves.
• Its helpful to complete one side at a time before moving to the next side of the arena.

7. Press firmly for 30-60 sec.
• It’s helpful to apply even pressure to the area with the applied MEK. This can be done by pressing the scrap acrylic on the area in need of pressure
8. Add another layer of MEK on the same side and apply pressure for 30-60 sec.
9. Repeat steps 6-8 for the remaining sides of the frame.
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Some sides might need more than two layers of MEK, so its important to make sure the wall piece is in place by tapping it slightly to see where more MEK is needed

10. Apply 70% ethanol to a Kimwipe and wipe the inside of the half-constructed arena and the side of the top piece with the groove.
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It’s important to thoroughly clean the pieces before bonding the piece to the frame. Any remaining dust or contaminants will not be able to be removed after fabrication.

11. Place the top piece on the lab bench with grove side up.
12. Line up the half-constructed arena with the top piece, making sure the frame is aligned toward the bench top and the sides line up to the grooves on the top piece.

13. Press down firmly over the whole arena and start applying MEK to one of the sides.
• Its’ helpful to angle the pipette downward instead of straight into the groove so the MEK travels to the grooves.
• Apply MEK to all sides of the arena doing one at a time and applying pressure for 30-60 sec.

14. Once completed, tape gets applied to the arena, covering the miracidia insert hole and the agar hole.
• The tape gets labeled with the date and your initials

15. To ensure that the MEK sealed the edges, the arena gets placed under something heavy (like a bottle of solution for example) for 5 min