From CLARITY Wiki
Chamber construction procedure
Mark the plastic container
Screw down the cap of a 60 mL Nalgene container tightly. While tightly shut, mark the areas to be soldered for the inlet (bottom) and outlet (top) fittings, making sure that they are on the same plane and on opposite sides of the container. Also, mark the spots to be soldered for the electrode fittings. The spots should be near the bottom of the chamber on the opposite side of the inlet fitting. One electrode fitting should be placed on either side of the inlet with plenty of room to fit a sample holder between the two electodes. Mark the top and bottom of the container with some type of symbol or initials in order to be able to identify matching caps and bottoms later on (if making more than one chamber).
Solder holes for fittings
Solder holes into the marked areas of the Nalgene container. Once the hole is big enough, screw in the inlet/outlet fitting while the plastic is still hot and melted. The fitting does not need to be fully screwed in, but it should be inserted just enough such that it cannot be easily pulled out by tugging. Solder the holes for the electrode fittings, and insert the smaller end of the electrode fitting into the container while the plastic is still melted.
Add epoxy to seal
Apply epoxy around the junctures of the inlet/outlet fittings on both the inside and outside of the container. Avoid getting epoxy into the inside of the fitting so that the inlet/outlet does not seal shut. Epoxy only the outside squares of the electrode fittings to the container, making sure to leave the fitting interior clear for electrode insertion. Use a slow-drying epoxy to allow seepage into the junctures and let dry overnight with each epoxy application.
Construct platinum wire electrodes
Fabricate electrodes out of 0.5 mm thick platinum wire using electric pliers, taking care to make both electrodes as identical as possible. Avoid bending and straightening the platinum wire over and over as it will eventually break. Insert the platinum electrodes delicately into the electrode fittings and use the pliers to adjust their positioning so that they are parallel to each other. Ensure that the electrodes are spaced to fit the sample holder snuggly (approximately the width of a 50 mL conical tube top). Apply epoxy to fill the dead space inside the electrode fittings and let dry overnight.
Cover with foil (if desired)
If you are clearing samples with fluorophores, aluminum foil tape can be applied onto the outside surface of the chamber to lightproof it.
- Push-to-connect fittings for better leakage control
- Banana plug fittings for electrodes to prevent platinum wire breakage from alligator clips
- 3-D printed
- 3-D ceramic
- 125 mL Nalgene for larger tissues