Automated parallel capillary electrophoresis system with hydrodynamic sample injection

• US 20020040850A1
• Filed: 12/11/2001
• Published: 04/11/2002
• Est. Priority Date: 08/31/1999
• Status: Active Grant

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Spectrumedix Corporation
• Inventors: Kane, Thomas E., Li, Qingbo, Liu, Changsheng
• Granted Patent: US 6,953,521 B2
• Time in Patent Office: Days
• US Class Current: 204/453
• CPC Class Codes: C07K 1/26 Electrophoresis
• CPC Class Codes: G01N 27/44704 Details; Accessories
• CPC Class Codes: G01N 27/44743 Introducing samples
• CPC Class Codes: G01N 27/44782 of a plurality of samples
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

An automated capillary zone electrophoretic system is disclosed. The system employs a capillary cartridge having a plurality of capillary tubes. The cartridge has a first array of capillary ends projecting from one side of a plate. The first array of capillary ends are spaced apart in substantially the same manner as the wells of a microtitre tray of standard size. This allows one to simultaneously perform capillary electrophoresis on samples present in each of the wells of the tray. The system includes a stacked, dual carrousel arrangement to eliminate cross-contamination resulting from reuse of the same buffer tray on consecutive executions from electrophoresis. The system also has a container connected to the detection end of the capillaries. The container is provided with valving which facilitate cleaning the capillaries, loading buffer into the capillaries, introducing samples to be electrophoresced into the capillaries, and performing capillary zone electrophoresis on the thus introduced samples.

Claims

1. A capillary zone electrophoresis subsystem configured to cooperate with a light source and a light detector to detect migrating samples, said subsystem comprising:

a fluid container;

a plurality of capillary tubes, each capillary tube having a first end and a second end, the first ends being arranged to receive samples thereinto and the second ends terminating at a first level in the fluid container;

a power supply configured to apply a voltage across the first and second ends of the capillary tubes;

a pump connected to said fluid container via a pump conduit, said pump configured to introduce a liquid into the container when said pump conduit is open and said pump is operating;

a vacuum device connected to said fluid container via a vacuum conduit entering said fluid container at a second level higher than said first level, said vacuum device configured to cause a negative pressure in said container, when said pump conduit is closed and said container is sealed;

a gas release valve connected to said container and configured to vent a gas in the container when said gas release valve is opened; and

a drain valve connected to said container and configured to drain a liquid in said container, when said drain valve is open.

2. The capillary zone electrophoresis subsystem of claim 1, further comprising:

an overflow conduit connected to said container, said overflow conduit configured to release liquid held within the container, when a height of said liquid within the container exceeds a predetermined level.

3. The capillary zone electrophoresis subsystem of claim 2, further comprising:

an overflow valve positioned in said overflow conduit, said overflow valve having at least a first, open position, and a second, closed position.

4. The capillary zone electrophoresis subsystem of claim 1, wherein said vacuum device is a syringe.

5. The capillary zone electrophoresis subsystem of claim 1, wherein said capillary first ends are arranged in a two-dimensional array having a spacing corresponding to that of wells of a microtitre tray, said system further comprising:

a positioning apparatus comprising an upper and a lower carrousel carrying microtitre trays, said positioning apparatus arranged to position one of said microtitre trays such that said two dimensional array of capillary first ends is inserted into corresponding wells of said microtitre tray.

6. The capillary zone electrophoresis subsystem of claim 1, wherein the vacuum device and the pump are separate devices.

7. A capillary zone electrophoresis subsystem configured to cooperate with a light source and a light detector to detect migrating samples, said subsystem comprising:

a fluid container;

a plurality of capillary tubes, each capillary tube having a first end and a second end, the first ends being arranged to receive samples thereinto and the second ends terminating at a first level in the fluid container;

a power supply configured to apply a voltage across the first and second ends of the capillary tubes;

a pump connected to said fluid container via a pump conduit, said pump configured to introduce a liquid into the container when said pump conduit is open and said pump is operating; and

a vacuum device connected to said fluid container via a vacuum conduit entering said fluid container at a second level higher than said first level, said vacuum device configured to cause a negative pressure in said container, when said pump conduit is closed and said container is sealed, wherein the vacuum device and the pump are separate devices.

8. The capillary zone electrophoresis subsystem of claim 7, further comprising:

a gas release valve connected to said container and configured to vent a gas in the container when said gas release valve is opened;

9. The capillary zone electrophoresis subsystem of claim 8, further comprising:

a drain valve connected to said container and configured to drain a liquid in said container, when said drain valve is open.

10. The capillary zone electrophoresis subsystem of claim 9, further comprising:

an overflow conduit connected to said container, said overflow conduit configured to release liquid held within the container, when a height of said liquid within the container exceeds a predetermined level.

11. The capillary zone electrophoresis subsystem of claim 10, further comprising:

an overflow valve positioned in said overflow conduit, said overflow valve having at least a first, open position, and a second, closed position.

12. The capillary zone electrophoresis subsystem of claim 7, wherein said capillary first ends are arranged in a two-dimensional array having a spacing corresponding to that of wells of a microtitre tray, said system further comprising:

a positioning apparatus comprising an upper and a lower carrousel carrying microtitre trays, said positioning apparatus arranged to position one of said microtitre trays such that said two dimensional array of capillary first ends is inserted into corresponding wells of said microtitre tray.

13. The capillary zone electrophoresis subsystem of claim 7, wherein said vacuum device is a syringe.

14. The capillary zone electrophoresis subsystem of claim 7, wherein said vacuum device is a drain conduit provided with a valve member positioned at a height below a height of said capillary first ends, whereby a gravity flow of liquid through said drain conduit by gravity causes a negative pressure in said container, thereby siphoning samples into each of said plurality of capillary tube first ends.

16. The method according to claim 15, wherein the step of applying a negative pressure comprises:

withdrawing air in said liquid container at a point above a level of said liquid while said container is sealed, thereby causing samples to enter said capillary first ends.

17. The method according to claim 16, wherein the step of withdrawing air comprises:

retracting a plunger of a syringe connected to said liquid container.

18. The method according to claim 15, wherein said step of applying a negative pressure comprises:

draining a portion of a liquid in said liquid container, such that samples are siphoned.

19. The method according to claim 15, further comprising:

flowing a liquid past said capillary second ends while applying said voltage differential.

20. The method according to claim 19, wherein the step of applying a negative pressure comprises withdrawing air in said liquid container at a point above a level of said liquid while said container is sealed, thereby causing samples to enter said capillary first ends.

21. The method according to claim 20, wherein the step of withdrawing air comprises:

retracting a plunger of a syringe connected to said liquid container.

22. The method according to claim 19, wherein the step of applying a negative pressure comprises draining a portion of a liquid in said liquid container, such that samples are siphoned.

15. A method for capillary zone electrophoresis on a plurality of samples, said method comprising:

providing a fluid container;

providing a plurality of capillary tubes, each capillary tube having a first end and a second end, the first ends being arranged to receive samples thereinto and the second ends terminating at a first level in the fluid container;

introducing a first liquid into said fluid container to a height at least as high as said first level;

applying a negative pressure to said capillary second ends to hydrodynamically introduce a sample to be electrophoresced into each of said capillary first ends, wherein the step of applying a negative pressure is performed by a device distinct from that used for the step of introducing a first liquid; and

applying a voltage differential between said capillary first ends and said capillary second ends to cause said samples to migrate towards said capillary second ends through electro-osmotic flow.