Dielectric and conductive imaging applied to gel electrophoresis

• US 7,767,071 B1
• Filed: 02/16/2006
• Issued: 08/03/2010
• Est. Priority Date: 02/16/2005
• Status: Expired due to Fees

• Current Assignee: Lloyd Douglas Clark
• Original Assignee: Lloyd Douglas Clark
• Inventors: Clark, Lloyd Douglas
• Primary Examiner(s): Neckel; Alexa D
• Assistant Examiner(s): DIETERLE, JENNIFER M
• Application Number:
  US11/355,549
• Time in Patent Office: 1,629 Days
• Field of Search: 204450-477, 204600-650, 204400-435
• US Class Current: 204/600
• CPC Class Codes: G01N 27/4473 by electric means

Abstract

A system for displaying the positions of substances in electrophoretic separations comprises a gel (635) a substance to be separated into a region (450), plates (400, 401) with electrodes (405-420 and 430-445) placed on either side of the gel. A source (315) applies a voltage to an electrode on one side of the gel, and an electrode on the other side of the gel is connected to the input of an amplifier (320). Multiplexers (605, 610) connect the source and amplifier to electrodes on either side of the gel. A computer (600) issues commands to the multiplexers to connect the source and amplifier to individual electrodes, and while a pair of electrodes is so connected, the computer activates a sample-and-hold circuit (620) and an A/D converter (630) to record the value of the electrical impedance within the gel at the location of the selected electrodes. The impedance value is stored in the computer'"'"'s memory and also displayed on a monitor (635) connected to the computer. The result is visualization of a gel-electrophoretic separation without requiring the use of stains, radioactivity, or molecular tags.

Claims

1. A system for detecting and displaying two-dimensional images of electrophoretic separations of constituents of a substance in a gel said constituents being capable of changing the electrical impedance within said gel, comprising:

a computer,

a gel having first and second sides,

a first insulating plate fitted with a first plurality of parallel, linear electrodes adjacent said first side of said gel,

a second insulating plate fitted with a second plurality of parallel, linear electrodes adjacent said second side of said gel, said first plurality of linear electrodes adjacent said first side of said gel being oriented orthogonally with respect to said second plurality linear electrodes adjacent said first side of said gel so that at least one of said electrodes adjacent said second side of said gel overlaps at least one of said electrodes adjacent said second side of said gel,

said first and said second insulating plates being fixed in position relative to one-another during use and separable at other times,

a voltage source arranged to be connectable sequentially and capable of applying a voltage to first plurality of parallel, linear electrodes adjacent said first side of said gel,

an amplifier arranged to be connectable sequentially to second plurality of parallel, linear electrodes adjacent said second side of said gel and capable of sensing changes in electrical impedance at a location where said first and second plurality of electrodes overlap and reporting said changes to said computer,

whereby said changes in electrical impedance can be recorded and displayed by said computer.

2. The system of claim 1, wherein at least one of said first and second pluralities of electrodes is in contact with said gel.

3. The system of claim 1, further including a first insulating layer interposed between said gel and said first plurality of electrodes on said first insulating plate.

4. The system of claim 1, further including a second insulating layer interposed between said gel and said second plurality of electrodes on said second insulating plate.

5. The system of claim 1, wherein said electrophoretic separation is selected from the group consisting of a separation in progress and a completed separation.

6. The system of claim 1, having an output provided on a plurality of humanly sensible indicia selected from the class consisting of visual displays, sounds, and printouts.

7. A method for imaging electrophoretic separations of a substance in a gel where said substance contains a plurality of separable constituents, comprising:

providing a gel having first and second sides and prepared for a separation,

providing a substance for separation,

providing a first insulating plate fitted with a plurality of parallel, linear electrodes adjacent said first side of said gel,

providing a second insulating plate fitted with a plurality of parallel, linear electrodes adjacent said second side of said gel, said electrodes adjacent said second side of said gel being oriented orthogonally with respect to said electrodes adjacent said first side of said gel so that at least one of said electrodes adjacent said first side of said gel overlaps at least one of said electrodes adjacent said second side of said gel,

said first and said second insulating plates being fixed in position relative to one-another during use and separable at other times,

providing a voltage source,

applying voltage from said source sequentially to each of said plurality of parallel, linear electrodes adjacent said first side of said gel,

providing an amplifier arranged to sense changes in electrical impedance where said electrodes adjacent said first side of said gel and said electrodes adjacent said second side of said gel overlap,

connecting said amplifier sequentially to each of said electrodes adjacent said second side of said gel so that at least one of said electrodes adjacent said second side of said gel overlaps at least one of said electrodes adjacent said first side of said gel,

providing a computer arranged to receive, record, and display said changes in said electrical impedance,

substance into said constituents within said gel, and

sensing, recording, and displaying differences in electrical properties between said gel in the presence of said constituents and said gel in the absence of said constituents, and displaying the locations of said constituents in said substance within said gel,

whereby said electrophoretic separation of said substance is imaged.

8. The method of claim 7, further including separating said substance into said constituents and detecting and displaying the loci of said constituents in said substance after the process of separating is complete.

9. The method of claim 7, further including providing an insulator over at least one of said first plurality of electrodes on said first insulating plate, one of said second plurality of electrodes on said second insulating plate, or both.

10. The method of claim 7, further including displaying the locations of said constituents in said substance during the process of separation.

11. The method of claim 7, further including storing a first set of data comprising the electrical properties of said gel before said separation, then storing a second set of data comprising the electrical properties of said gel after said separation, and comparing said first and said second sets of data.

12. Apparatus for detecting, recording, and displaying one-dimensional and two-dimensional images of electrophoretic separations of migratory materials in a gel, comprising:

a gel having first and second sides and containing said migratory materials,

a first insulating plate fitted with a first plurality of parallel, linear electrodes adjacent said first side of said gel,

a second insulating plate fitted with a second plurality of parallel, linear electrodes adjacent said second side of said gel, said second plurality of linear electrodes adjacent said second side of said gel being oriented orthogonally with respect to said first plurality of linear electrodes adjacent said first side of said gel,

said first and said second insulating plates being fixed in position relative to one-another during use and separable at other times,

a voltage source sequentially connected to said first plurality of electrodes on said first plate by a first multiplexing means,

a sample-and-hold means,

an analog-to-digital converter means,

an amplifier sequentially connected to said first plurality of electrodes on said second plate by a second multiplexing means, said amplifier, in combination with said sample-and-hold means and said analog-to-digital converter means, being capable of detecting differences in electrical properties between said gel and said materials for each pair of electrodes selected by said first and said second multiplexing means,

a computer arranged to control said voltage source, said sample and hold means, said analog-to-digital converter means, and said first and said second multiplexing means and to monitor, report, and display differences in said electrical properties between said separated materials within said gel,

whereby said differences in said electrical properties representative of said separations are detected, recorded, and displayed by said computer.

13. The apparatus of claim 12, wherein at least one of said first plurality or said second plurality of electrodes is in contact with said gel.

14. The apparatus of claim 12, further including an insulating layer interposed between at least one of said first plurality of said second plurality or electrodes and said gel.

15. The apparatus of claim 12, further including providing an output on a plurality of humanly sensible indicia selected from the class consisting of visual displays, sounds, and printouts.