Capillary array electrophoresis scanner

• US 6,270,644 B1
• Filed: 01/27/1999
• Issued: 08/07/2001
• Est. Priority Date: 01/27/1999
• Status: Expired due to Term

• Current Assignee: Affymetrix, Inc. (Thermo Fisher Scientific Incorporated)
• Original Assignee: Affymetrix, Inc. (Thermo Fisher Scientific Incorporated)
• Inventors: Mathies, Richard A., Scherer, James R.
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): NOGUEROLA, ALEXANDER STEPHAN
• Application Number:
  US09/238,131
• Time in Patent Office: 923 Days
• Field of Search: 356/344, 204/451, 204/452, 204/601, 204/603, 422/70
• US Class Current: 204/603
• CPC Class Codes: G01N 27/44721 by optical means
• CPC Class Codes: G01N 27/44782 of a plurality of samples

Abstract

A Monster Capillary Array Electrophoresis scanner measures four-color electropherograms from over a thousand capillary electrophoretic separations in parallel. The system consists of a two-dimensional confocal rotary scanner and a four-color detection unit.

Claims

1. A capillary array electrophoresis apparatus comprising:

two cylinders, each having a side;

a plurality of capillary tubes disposed in a circular array on the sides of the two cylinders;

a source of radiant energy of a first wavelength;

a rotor means for rotating in a single direction;

an objective lens, mounted on the rotor means, for receiving and focusing the radiant energy of a first wavelength onto the capillary tubes;

a dichroic beam splitter mounted on the rotor means;

confocal spatial and spectral filter means for transmitting radiant energy of a second wavelength emitted from the capillaries and rejecting radiant energy of other wavelengths; and

,

a detector means for measuring the intensity of the second radiant energy.

2. The capillary array electrophoresis apparatus of claim 1, further comprising:

computer means for process control, data processing, and for controlling the rotor means.

3. The capillary array electrophoresis apparatus of claim 1, wherein the rotor means is connected to a micro-stepping indexed motor.

4. The capillary array electrophoresis apparatus of claim 1, wherein said detector means is a four-color detection means incorporating photomultiplier means.

5. The capillary array electrophoresis apparatus of claim 1, wherein the radiant energy of a first wavelength is passed through a quarter-wave plate calibrated at about 488 nM.

6. The capillary array electrophoresis apparatus of claim 1, further comprising a photodiode positioned between the objective lens and a capillary, wherein the radiant energy of a first wavelength initiates data acquisition by illuminating the photodiode.

7. The capillary array electrophoresis apparatus of claim 1, wherein said tubes are mounted in grooves on a side of said cylinders.

8. The capillary array electrophoresis apparatus of claim 1, wherein said tubes are substantially adjacent to one another.

9. The capillary array electrophoresis apparatus of claim 1, incorporating at least about 1024 of said tubes.

10. The capillary array electrophoresis apparatus of claim 1, wherein the source of radiant energy of a first wavelength is a coherent light source.

11. The capillary array electrophoresis apparatus of claim 10, wherein the coherent light source is a laser.

12. The capillary array electrophoresis apparatus of claim 1, wherein the detector means is a two-dimensional image array detector.

13. The capillary array electrophoresis apparatus of claim 12, wherein the array detector is selected from a group consisting of a charge-coupled device and a charge-injection device.

14. The capillary array electrophoresis apparatus of claim 1, wherein the cylinders are connected to one another by at least three posts.

15. An apparatus for determining the sequence of a nucleic acid sample, the apparatus comprising:

two cylinders, each having a circumference;

a plurality of electrophoresis lanes disposed along the circumference of the cylinders in a circular array;

a source of radiant energy of a first wavelength;

an optical detection system for detecting radiant energy of a second wavelength emanating from the electrophoresis lanes, the optical detection system comprising;

(i) collection optics for collecting and focusing the radiant energy of a second wavelength;

(ii) a detector for measuring the intensity of the radiant energy of a second wavelength; and

,

(iii) a rotary scanner for directing the radiant energy of a first wavelength to each electrophoresis lane.

16. The apparatus of claim 15, wherein the rotor means is connected to a micro-stepping indexed motor.

17. The apparatus of claim 15, wherein the detector means is a four-color detection means incorporating photomultiplier means.

18. The apparatus of claim 15 wherein the source of radiant energy of a first wavelength is a coherent light source.

19. A capillary array electrophoresis apparatus, the apparatus comprising:

a source of radiant energy of a first wavelength;

at least one cylinder having a circumference;

a rotor;

an objective lens, mounted on the rotor and housed within the at least one cylinder, for focusing the radiant energy of a first wavelength, wherein the objective lens is positioned in the path of the radiant energy;

a plurality of capillaries positioned along the circumference of the at least one cylinder and in the path of the radiant energy of a first wavelength focused by the objective lens; and

at least one detector for detecting radiant energy of a second wavelength emitted from the capillaries.

20. The capillary array electrophoresis apparatus of claim 19, further comprising a photodiode positioned between the objective lens and a capillary such that illumination of the photodiode initiates detection by the detector.

21. A capillary array electrophoresis apparatus, the apparatus comprising:

a source of radiant energy of a first wavelength;

three vertical posts;

two cylinders, each having a circumference, wherein the two cylinders share a common vertical axis and are mounted on the posts;

a rotor;

an objective lens, mounted on the rotor and housed within the at least one cylinder, for focusing the radiant energy of a first wavelength, wherein the objective lens is positioned in the path of the radiant energy;

a plurality of capillaries positioned along the circumference of the at least one cylinder and in the path of the radiant energy of a first wavelength focused by the objective lens;

four dichroic beam splitters, each calibrated to pass a different wavelength of radiant energy emitted from the capillaries and to reject all other wavelengths; and

four photodetectors, each of which measures the intensity of radiant energy passed to it by one of the beam splitters.