Polymeric electrode for electrophysiological testing
View Patent ↗A polymeric material such as PDMS is molded into an electrode structure containing a micron-size aperture for receiving and forming a giga-ohm seal with a biological membrane. One end of a tube is filled with uncured polymeric material and pressed against a support surface to prevent drainage. A conventional micropipette having a size suitable for sliding through the tube is introduced, tip first, into the tube and is allowed to fall through the polymeric material and rest against the support surface. The assembly is heated to cure the polymer and the micropipette is removed from the tube, thereby leaving a polymeric plug at the end of the tube with an aperture suitable in shape and size for patch-clamp giga-ohm seal electrode applications. A multi-well tray with a polymeric electrode plug in each well is constructed using the same approach.
1. A patch-clamp electrode structure comprising:
a support structure defining a cavity;
a polymeric plug in the cavity; and
an aperture through the polymeric plug;
wherein said aperture conforms to a pointed tip of a patch-clamp micropipette; and
wherein said cavity includes a retaining protrusion for anchoring the polymeric plug.
2. A multi-well tray manufactured according to the method comprising the following steps:
providing a tray with a plurality of wells and an electrode perforation in each well;
placing a predetermined amount of uncured polymeric material in the electrode perforation in each well to form a layer over a support surface;
piercing said layer in each well with a pointed plunger having a tip conforming to an aperture suitable for forming a high-resistance patch-clamp seal with a biological membrane;
curing the layer of polymeric material to form a solid plug within the electrode perforation; and
removing the pointed plunger and the support surface from the plug.
3. A multi-well patch-clamp electrode structure comprising:
a tray with a plurality of wells and an electrode perforation in each well;
a polymeric plug in the electrode perforation of each well; and
an aperture through each polymeric plug;
wherein said aperture conforms to a pointed tip of a patch-clamp micropipette.
4. The electrode structure of claim 3 , wherein said polymeric plug comprises a cured silicone polymer.
5. The electrode structure of claim 3 , wherein said polymeric plug comprises cured poly-dimethylsiloxane.
6. The electrode structure of claim 3 , further comprising a layer of polymeric material with a plurality of openings overlaying said electrode perforations; and wherein said polymeric plug extends into said overlaying opening in the layer of polymeric material.
7. The electrode structure of claim 3 , wherein said electrode perforation includes a retaining protrusion for anchoring the polymeric plug.
8. The electrode structure of claim 3 , wherein said aperture has a diameter of about 0.5-20 microns at a smaller end thereof.
9. The electrode structure of claim 3 , wherein said aperture is at least 0.1 mm deep.