High throughput screening of ion channels

摘要

Multi-well plates having contoured well designs allow multi-stage high throughput parallel assaying of ion channels or ion transporters. A well of a multi-well plate has a bottom region that is sized and shaped to simultaneously accommodate a sensing electrode and a pipette for delivering test compounds, wash fluid, and optionally ligands. The multi-well plates when coupled with an instrument having a pipette head and an electrode plate facilitates fluidic contact between cells and fluids delivered via a pipette for washing of wells with buffers or other wash solutions for serial exposure of test cells to various reagents or other stimuli. The control and test experiments are performed on the same cell (or cells) in a single well.

主权项

1. A method of conducting a patch clamp assay, the method comprising:
(a) providing a well wherein the well comprises a bottom having an electrode pocket sized and shaped to accommodate a sensing electrode and a cell cavity wherein, the cell cavity comprises a cell sealing aperture in a bottom surface of the cell cavity and a pipette guide sized and shaped for mating with a pipette tip; (b) introducing a first conductive solution to the well utilizing a pipette with a pipette tip; (c) exchanging the first conductive solution within the well immediately before delivering a cell to the well by dwelling the pipette tip in the pipette guide, wherein the pipette tip is dwelled in the pipette guide prior to delivering the cell to the well; (d) providing the cell positioned within the cell cavity and sealed to the aperture on the bottom surface of the cell cavity; (e) inserting the pipette via the pipette guide into the well with the pipette tip facing the cell, wherein the pipette tip is dwelled in the pipette guide; (f) measuring a first electrical signal from the cell at the time or after the cell is exposed to the first conductive solution and allowing for continuous control of a voltage applied to the cell, while maintaining the dwelled pipette tip within the pipette guide during the measuring and controlling operations for accommodating an outflow and preventing a backflow of the first conductive solution exposed to the cell; (g) repeating step (c); and (h) delivering an additional solution to the bottom of the well and/or replacing the first conductive solution within the well in steps (e) and (g) by gradually retracting the pipette from the pipette guide followed by advancing the pipette into the pipette guide and dwelling the pipette tip therein.