System for rapid chemical activation in high-throughput electrophysiological measurements

• US 20020014408A1
• Filed: 05/21/2001
• Published: 02/07/2002
• Est. Priority Date: 08/04/2000
• Status: Active Grant

• Current Assignee: Essen Instruments, Inc. (Sartorius AG)
• Original Assignee: Essen Instruments, Inc. (Sartorius AG)
• Inventors: Schroeder, Kirk S.
• Granted Patent: US 7,067,046 B2
• Time in Patent Office: Days
• US Class Current: 204/403.1
• CPC Class Codes: G01N 35/10 Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices G01N35/0099 takes precedence

Abstract

An ultraviolet light source is added to a high-throughput electrophysiogical measurement system to allow for rapid chemical stimulation via caged compound release to a plurality of measurement samples. The integrated electrophysiogical measurement system includes a computer-controlled data collection system, an integrated electronics head for making parallel electrical measurements, and an integrated fluidics head used in part to transfer test compounds into the measurement process. This light source, and associated light coupling to a plurality of test samples, is used in conjunction with the system to make effectuate high-throughput electrical measurements with respect to fast-acting, chemically-activated electrophysiological events. The UV-source modification allows for rapid stimulation and measurement of multiple fast ligand-gated ion channel events in parallel.

Claims

1. Electrophysiological measurement apparatus, comprising:

a multi-well plate having a plurality of fluid chambers, each containing biological material to be analyzed;

a thin substrate having an array of apertures in alignment with the chambers of the multi-well plate;

the substrate being bonded to the multi-well plate such that the chambers are open at the top and sealed at the bottom except for the apertures;

the apertures being smaller in diameter than the biological material, thereby enabling a high-resistance seal to be formed between the material in each chamber and a corresponding aperture;

a fluid plenum to receive the multi-well plate such that at least the substrate is immersed;

a first electrode disposed in the fluid plenum;

at least one second electrode moveable into the top openings of the fluid chambers of the multi-well plate;

an ultraviolet light source for illuminating one or more of the chambers so as to facilitate the rapid release of a caged compound present therein; and

electrophysiological measurement circuitry in electrical communication with the electrodes.

2. The electrophysiological measurement apparatus of claim 1, including a single aperture associated with each chamber of the multi-well plate.

3. The electrophysiological measurement apparatus of claim 1, wherein the substrate is a plastic substrate having a glass coating at least in the region where the high-resistance seal is formed between the material and the substrate.

4. The electrophysiological measurement apparatus of claim 1, wherein the substrate is mylar or polyimide.

5. The electrophysiological measurement apparatus of claim 1, wherein the apertures are in the range of 1 to 10 micrometers.

6. The electrophysiological measurement apparatus of claim 1, wherein the apertures are tapered.

7. The electrophysiological measurement apparatus of claim 1, wherein the multi-well plate is sealed to the fluid plenum, enabling a differential pressure to be applied relative to the fluid in each chamber, thereby causing the material in each chamber to migrate to a respective aperture.

8. The electrophysiological measurement apparatus of claim 1, wherein the fluid plenum includes a chemical reagent causing the material in each chamber to permeablize in the vicinity of the aperture.

9. The electrophysiological measurement apparatus of claim 1, further including:

a mechanism for moving the electrode into the chambers of the multi-well plate so as to automate the measurement of the material contained therein.

10. The electrophysiological measurement apparatus of claim 1, further including:

a plurality of electrodes in alignment with a plurality of the chambers of the multi-well plate; and

a mechanism for moving the electrodes into the chambers of the multi-well plate to perform simultaneous measurements on the material contained therein.

11. The electrophysiological measurement apparatus of claim 1, further including:

a system for transferring fluids from one or more sources to the chambers of the multi-well plate.

12. The electrophysiological measurement apparatus of claim 1, further including:

a mechanism for directing the light from the source to the multi-well plate in an automated and spatially selectable manner.

13. The electrophysiological measurement apparatus of claim 1, further including:

a mechanism for directing the light from the source to spatially selected wells in conjunction with a plurality of electrodes, thereby providing for the simultaneous electrical recording of the biological samples during and subsequent to the ultraviolet illumination of the samples.

14. The electrophysiological measurement apparatus of claim 1, further including:

an electronic control of optical pulsewidth and intensity such that the timing, duration and light energy of the ultraviolet exposure can be automatically controlled.

15. The electrophysiological measurement apparatus of claim 1, further including:

a guide for coupling the ultraviolet light from the source directly into the biological samples.

16. The electrophysiological measurement apparatus of claim 1, wherein the guide includes one o r more optical fibers.

17. The electrophysiological measurement apparatus of claim 1, wherein the guide includes one or more mirrors, beam splitters, diffusers, collimators and telescopic optics.