High density electrophoresis device and method

• US 7,491,305 B2
• Filed: 12/08/2003
• Issued: 02/17/2009
• Est. Priority Date: 07/16/1999
• Status: Expired due to Fees

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems, Inc. (Thermo Fisher Scientific Incorporated)
• Inventors: Kowallis, Reid B., Woudenberg, Timothy M.
• Primary Examiner(s): Olsen; Kaj K
• Application Number:
  US10/731,420
• Publication Number:
  US 20040178074A1
• Time in Patent Office: 1,898 Days
• Field of Search: 204/600, 204/601, 204/606, 204/450, 204/451, 204/456, 494/33, 494/81, 494 13- 14, 422/72, 422/82.01
• US Class Current: 204/601
• CPC Class Codes: B01L 2200/0684 Venting, avoiding backpressure, avoid gas bubbles
• CPC Class Codes: B01L 2300/044 pierceable, e.g. films, membranes
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0803 Disc shape
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/12 Specific details about materials
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B01L 3/502723 characterised by venting arrangements
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 2565/629 being a microfluidic device
• CPC Class Codes: G01N 27/44721 by optical means
• CPC Class Codes: G01N 27/44782 of a plurality of samples
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: G01N 35/00069 : whereby the sample substrate is of the bio-disk type, i.e. having the format of an optical disk

Abstract

The present invention provides, in one aspect, an apparatus or electrophoretic separation of analytes. In one aspect, the apparatus comprises a disc-shaped substrate defining (1) a central reservoir region, (2) a plurality of electrophoretic channels in fluid communication with, and emanating substantially radically from, the central reservoir region, the channels being coplanar with each other, and each channel having (i) a proximal end which is linked to the reservoir region, and (ii) a distal end, and preferably (3) for each channel, at least one chamber, and preferably three chambers, linked by a passageway in fluid communication with the distal end of that channel. Preferably, each passageway leads from each chamber in a direction that is initially away from the central reservoir region, whereby centrifugation of the substrate about a central axis that is perpendicular to the channels is effective to disperse liquid from the central reservoir region into the channels and chambers, such that any air bubbles in the chambers, channels, and passageways are forced towards the axis of rotation, when such liquid is present in the central reservoir region.

Claims

1. An analytical system comprising:

a substrate including;

a central reservoir region,

a plurality of electrophoretic channels in fluid communication with, and emanating substantially radically from, the central reservoir region, the channels being coplanar with each other, and each channel having (i) a proximal end which is linked to the central reservoir region, and (ii) a distal end,

a first chamber that is linked by a first passageway in fluid communication with the distal end of a first of the plurality of eleetrophoretic channels, and

a second chamber that is linked by a second passageway in fluid communication with the distal end of a second of the plurality of electrophoretic channels;

a mechanism for rotating the substrate about a central axis of rotation that is perpendicular to the plane of the channels; and

a plurality of electrodes for applying a voltage potential between the first chamber and the central reservoir region and between the second chamber and the central reservoir region.

2. The analytical system of claim 1, whereby centrifugation of the substrate about the central axis is effective to disperse liquid from the central reservoir region into the channels and the first and second chambers such that any air bubbles in the first and second chambers, the channels, and the passageways are forced towards the axis of rotation, when such liquid is present in the central reservoir region.

3. The analytical system of claim 1, further comprising a contact card adapted to supply separate electrical voltages to the electrodes.

4. The analytical system of claim 1, further comprising:

conductive concentric rings in electrical contact with the electrodes; and

conductive brushes which remain in contact with the concentric rings, when the substrate is rotated.

5. The analytical system of claim 1, further comprising a voltage source adapted to independently control each of the plurality of electrodes.

6. The analytical system of claim 1, further comprising a detector for detecting selected components which may be present in one or more of the channels.

7. The analytical system of claim 6, wherein the mechanism for rotating the substrate about the central axis is adapted to sequentially pass the channels by the detector, for detecting one or more components that may be present in the channels.

8. The analytical system of claim 1, wherein said chambers are defined in part by an annular septum that covers the chambers and permits needle-access to the chambers for delivery of liquid to the chambers.

9. The analytical system of claim 1, wherein said chambers are defined in part by an annular cover that is porous to air, such that displaced air escapes through the annular cover when liquid is loaded into the channels.

11. The analytical system of claim 1, wherein the channels have cross-sectional diameters between 1 μ

m and 100 μ

m.

12. The analytical system of claim 1, wherein the substrate comprises a material comprising at least one of copper, aluminum, glass, silica-based glass, quartz, and polycarbonate.

13. The analytical system of claim 1, wherein the substrate further comprises electrical resistive traces in thermal contact with the first and second chambers of each channel.

14. The analytical system of claim 1, wherein the mechanism comprises a motor shaft to rotate the substrate, and wherein the motor shaft is electrically grounded and adapted to be in communication with a liquid disposed in the central reservoir region.

10. The analytical system of clalm 1, wherein at least one of the channels contains an electrophoresis medium.