Multiple capillary biochemical analyzer

• US 5,741,412 A
• Filed: 10/23/1996
• Issued: 04/21/1998
• Est. Priority Date: 06/03/1993
• Status: Expired due to Fees

• Current Assignee: University of Alberta
• Original Assignee: University of Alberta
• Inventors: Zhang, Jian Zhong, Dovichi, Norman J.
• Primary Examiner(s): Gorgas, Kathryn L.
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US08/735,540
• Time in Patent Office: 545 Days
• Field of Search: 204/451, 204/452, 204/453, 204/454, 204/455, 204/601, 204/602, 204/603, 204/604, 204/605
• US Class Current: 204/602
• CPC Class Codes: G01N 15/14 Optical investigation techniques, e.g. flow cytometry
• CPC Class Codes: G01N 27/44721 by optical means
• CPC Class Codes: G01N 27/44782 of a plurality of samples
• CPC Class Codes: G01N 30/466 with separation columns in parallel
• CPC Class Codes: G01N 30/74 Optical detectors measurement of intensity, velocity, spectral content, polarisation, or phase of infrared, visible or ultraviolet light G01J

Abstract

A multiple capillary analyzer allows detection of light from multiple capillaries with a reduced number of interfaces through which light must pass in detecting light emitted from a sample being analyzed, using a modified sheath flow cuvette. A linear or rectangular array of capillaries is introduced into a rectangular flow chamber. Sheath fluid draws individual sample streams through the cuvette. The capillaries are closely and evenly spaced and held by a transparent retainer in a fixed position in relation to an optical detection system. Collimated sample excitation radiation is applied simultaneously across the ends of the capillaries in the retainer. Light emitted from the excited sample is detected by the optical detection system. The retainer is provided by a transparent chamber having inward slanting end walls. The capillaries are wedged into the chamber. One sideways dimension of the chamber is equal to the diameter of the capillaries and one end to end dimension varies from, at the top of the chamber, slightly greater than the sum of the diameters of the capillaries to, at the bottom of the chamber, slightly smaller than the sum of the diameters of the capillaries. The optical system utilizes optic fibres to deliver light to individual photodetectors, one for each capillary tube. A filter or wavelength division demultiplexer may be used for isolating fluorescence at particular bands.

Claims

1. An analyzer for analyzing an organic sample, the analyzer comprising:

a plurality of capillary tubes arrayed side by side, each capillary tube having first and second ends, the second ends of the capillary tubes terminating adjacent each other and the first ends being connectable to a source of organic sample;

a flow chamber having an interior cavity, the second ends of the capillary tubes terminating inside the interior cavity;

means to supply sheath fluid into the interior cavity of said flow chamber to provide a flow of sheath fluid past the second ends of the capillary tubes such that any organic sample in said capillary tubes is drawn by the flow of sheath fluid in individual sample streams from the second ends of the capillary tubes;

means to force said organic sample through the capillary tubes from the first ends of the capillary tubes to the second ends of the capillary tubes;

a detector positioned to detect organic sample in the individual sample streams emerging from the capillary tubes; and

a non-capillary sized waste outlet, independent of said means to force organic sample, positioned in said flow chamber for draining said sheath flow fluid and entrained sample streams directly from said flow chamber to waste, said waste outlet draining the entrained sample streams and associated sheath flow fluid from a plurality of said capillary tubes.

2. The analyzer according to claim 1, wherein the detector includes a source of collimated electromagnetic radiation having a wavelength for exciting the sample to emit radiation, said source being aligned to provide collimated light through the organic sample drawn from the capillary tubes by the sheath fluid.

3. The analyzer according to claim 2, wherein the detector further includes an optical system aligned to receive emitted radiation from the organic sample.

4. The analyzer according to claim 3, in which the optical detection system further comprises a wave length division demultiplexer for separating emitted radiation into light of at least two spectral bands, and at least two photodetectors for each capillary tube, one for each of said spectral bands.

5. The analyzer according to claim 2, in which said capillary tubes are arrayed in a linear array.

6. The analyzer according to claim 2, wherein the detector comprises an optical system for receiving scattered radiation from contaminants carried by said sample.

7. The analyzer according to claim 2, wherein said flow chamber includes an upper portion and a lower portion, and said means to supply sheath fluid comprises a tube connected to the upper portion of the flow chamber and wherein the lower portion of said flow chamber encircles the second ends of the capillary tubes.

8. The analyzer according to claim 7, wherein at least a portion of the lower portion of the chamber is made of transparent material, said portion being adjacent the second ends of the capillary tubes.

9. The analyzer according to claim 2, wherein a fluid acceleration region is formed downstream of the second ends of said capillaries, and wherein said source of collimated electromagnetic radiation is positioned to direct said radiation at the fluid acceleration region.

10. The apparatus according to claim 2, wherein said detector further comprises an optical collection means positioned to collect radiation emitted by said sample in a direction generally orthogonal to the direction of flow of said sample streams.

11. The analyzer according to claim 1, wherein the means to force the organic sample through the capillary tubes includes an electrophoretic voltage source connected across the length of the capillary tubes.

12. The analyzer according to claim 1, wherein said capillary tubes are arrayed in a linear array.

13. The analyzer according to claim 1, wherein the means to supply sheath flow into said interior cavity includes a reservoir of sheath fluid, syphon coupled to the flow chamber.

14. The apparatus according to claim 13, wherein said detector further comprises a source of collimated electromagnetic radiation for generating electromagnetic radiation in a first direction which is generally orthogonal with respect to the direction of flow of said sample streams and an optical collection means positioned to receive radiation emitted from said sample streams in a second direction which is generally orthogonal with respect to the direction of flow of said sample streams.

15. The analyzer according to claim 1, wherein said flow chamber comprises a side wall at least partially encircling said interior cavity and wherein at least a segment of the side wall being transparent to electromagnetic radiation radiating away from organic sample drawn from the capillary tubes by the sheath fluid.

16. The analyzer according to claim 1, wherein said chamber comprises a side wall at least partially encircling said interior cavity and wherein at least a segment of the side wall being transparent to electromagnetic radiation radiating away from organic sample drawn from the capillary tubes by the sheath fluid and at least a further segment of the side wall being transparent to the collimated electromagnetic radiation.

17. The apparatus according to claim 16, wherein said detector further comprises a source of collimated electromagnetic radiation for generating electromagnetic radiation in a first direction which is generally orthogonal with respect to the direction of flow of said sample streams and an optical collection means positioned to receive radiation emitted from said sample streams in a second direction which is generally orthogonal with respect to the direction of flow of said sample streams.

18. The analyzer according to claim 1, wherein each of said capillary tubes has an inside diameter of about 50 μ

m and an outside diameter on the order of about 150 μ

m.

19. The analyzer according no claim 1, wherein said means to force an organic sample through the capillary tube comprises an electrophoretic voltage source of about 30 kv connected across said capillary tubes and said flow chamber.

20. The analyzer according to claim 19, further comprising means for distributing forces between capillaries generated by said voltage source.

21. The analyzer according to claim 1, wherein the organic material is carried in said capillaries by a fluid, said fluid having essentially the same index of refraction as the sheath fluid.

22. The analyzer according to claim 1, wherein the volumetric flow of sheath fluid through said chamber is less than about 10 mL/hr.

23. The analyzer according to claim 1, wherein the rate flow of sheath fluid through said chamber is less than about 4 mm/sec.

24. The apparatus according to claim 1, wherein said detector further comprises a source of collimated electromagnetic radiation for generating electromagnetic radiation in a first direction which is generally orthogonal with respect to the direction of flow of said sample streams and an optical collection means positioned to receive radiation emitted from said sample streams in a second direction which is generally orthogonal with respect to the direction of flow of said sample streams.