Multiplexed fluorescence detector system for capillary electrophoresis

• US 5,498,324 A
• Filed: 04/06/1994
• Issued: 03/12/1996
• Est. Priority Date: 02/05/1993
• Status: Expired due to Term

• Current Assignee: Iowa State University Research Foundation Incorporated (Iowa State University)
• Original Assignee: Iowa State University Research Foundation Incorporated (Iowa State University)
• Inventors: Taylor, John A., Yeung, Edward S.
• Primary Examiner(s): Niebling, John
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US08/223,864
• Time in Patent Office: 706 Days
• Field of Search: 204/180.1, 204/299 R, 356/344
• US Class Current: 204/452
• CPC Class Codes: G01N 27/44721 by optical means

Abstract

A fluorescence detection system for capillary electrophoresis is provided wherein the detection system can simultaneously excite fluorescence and substantially simultaneously monitor separations in multiple capillaries. This multiplexing approach involves laser irradiation of a sample in a plurality of capillaries through optical fibers that are coupled individually with the capillaries. The array is imaged orthogonally through a microscope onto a charge-coupled device camera for signal analysis.

Claims

1. A multiplexed method of detecting the presence of macromolecules in samples present in a plurality of capillaries in capillary electrophoresis comprising:

(a) providing a buffer solution containing a fluorophore;

(b) introducing samples into a plurality of capillaries in a capillary electrophoresis system, each capillary containing the buffer; and

(c) substantially simultaneously irradiating the samples in the capillaries with coherent light to cause fluorescence.

2. The method of claim 1 wherein the coherent light is transmitted through optical fibers individually coupled to the capillaries.

3. The method of claim 1 wherein the step of substantially simultaneously irradiating comprises irradiating through the wall of each capillary.

4. A multiplexed method of detecting the presence of macromolecules in samples present in a plurality of capillaries in capillary electrophoresis, comprising the steps of:

(a) providing a buffer solution containing a fluorophore;

(b) introducing samples into a plurality of capillaries in a capillary electrophoresis system, each capillary containing the buffer;

(c) irradiating each sample in each capillary to cause fluorescence of the macromolecule; and

(d) substantially simultaneously detecting changes in fluorescent emission.

5. The method of claim 4 wherein the step of substantially simultaneously detecting comprises detecting fluorescent emission through the wall of each of the capillaries by the fluorescent species.

6. A multiplexed fluorescence detection system for use with capillary electrophoresis to detect target species in samples comprising:

(a) an array of at least two capillaries each having an intake end, an outflow end, an inside diameter of about 40-100 microns, and an annular wall wherein at least a portion of the wall is translucent to light having a wavelength of about 200-1,500 nm, the intake end being in fluid communication with a sample so that the sample is drawn into the capillary;

(b) means for irradiating each sample in each capillary to cause fluorescence of the target species; and

(c) means for substantially simultaneously detecting changes in fluorescent emission.

7. The detection system of claim 6 wherein the capillary array comprises at least about 100 capillaries.

8. The detection system of claim 6 wherein the means for irradiating comprises means for substantially simultaneously irradiating each sample in each capillary with coherent light to cause fluorescence.

9. The detection system of claim 8 wherein the means for substantially simultaneously detecting changes in fluorescent emission comprises a single detector.

10. A multiplexed fluorescence detection system for use with capillary electrophoresis to detect target species in samples comprising:

(a) an array of at least two capillaries each having an intake end, an outflow end, and an annular wall wherein at least a portion of the wall is translucent to light having a wavelength of about 200-1500 nm, the intake end being in fluid communication with a sample so that the sample is drawn into the capillary;

(b) means for irradiating each sample in each capillary to cause fluorescence of the target species; and

(c) a single detector for substantially simultaneously detecting changes in fluorescent emission.

11. The detector system of claim 10 wherein the capillary array comprises at least about 100 capillaries.

12. The detection system of claim 10 wherein each capillary has an inside diameter of about 40-100 microns.

13. The detection system of claim 10 wherein the means for irradiating comprises means for substantially simultaneously irradiating each sample in each capillary with coherent light to cause fluorescence.

14. A multiplexed fluorescence detection system for use with capillary electrophoresis to detect target species in samples comprising:

(a) an array of at least two capillaries each having an intake end, an outflow end, and an annular wall wherein at least a portion of the wall is translucent to light having a wavelength of about 200-1,500 nm, the intake end being in fluid communication with a sample so that the sample is drawn into the capillary;

(b) an array of at least two optical fibers, each optical fiber having a first end and a second end, the second end of each optical fiber being coupled to a capillary in the array of capillaries;

(c) means for generating coherent light of a wavelength suitable to cause fluorescence in the target species present in the capillaries comprising collimating means positioned to distribute the coherent light onto the first ends of the optical fibers in a substantially uniform manner so as to substantially simultaneously irradiate the samples in the capillaries; and

(d) means for detecting changes in fluorescent emission.

15. A multiplexed fluorescence detection system for use with capillary electrophoresis to detect target species in samples comprising:

(a) an array of at least two capillaries each having an intake end, an outflow end, and an annular wall wherein at least a portion of the wall is translucent to light having a wavelength of about 200-1,500 nm, the intake end being in fluid communication with a sample so that the sample is drawn into the capillary;

(b) means for irradiating each sample in each capillary to cause fluorescence of the target species; and

(c) a charged-coupled device based imaging system for substantially simultaneously detecting changes in fluorescent emission.