Simultaneous analyte determination and reference balancing in reference T-sensor devices

• US 6,582,963 B1
• Filed: 10/31/2000
• Issued: 06/24/2003
• Est. Priority Date: 03/29/1996
• Status: Expired due to Term

• Current Assignee: Washington University In St Louis
• Original Assignee: University of Washington
• Inventors: Kenny, Margaret, Zebert, Diane, Wu, Caicai, Weigl, Bernhard H., Holl, Mark R.
• Primary Examiner(s): WALLENHORST, MAUREEN
• Application Number:
  US09/702,645
• Time in Patent Office: 966 Days
• Field of Search: 436/52, 436/53, 436/172, 436/177, 436/180, 422/81, 422/82, 422/82.08
• US Class Current: 436/52
• CPC Class Codes: B01F 25/31 in conduits or tubes through which the main component flows
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: B01F 33/3039 with mixing achieved by diffusion between layers
• CPC Class Codes: B01J 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01L 2200/0636 Focussing flows, e.g. to laminate flows
• CPC Class Codes: B01L 2200/148 Specific details about calibrations
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0457 passive flow or gravitation
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/502776 specially adapted for focusing or laminating flows
• CPC Class Codes: G01N 15/10 Investigating individual particles
• CPC Class Codes: G01N 15/14 Optical investigation techniques, e.g. flow cytometry
• CPC Class Codes: G01N 15/1404 Handling flow, e.g. hydrodynamic focusing
• CPC Class Codes: G01N 15/1484 microstructural devices
• CPC Class Codes: G01N 2035/00237 Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: G01N 30/0005 Field flow fractionation
• CPC Class Codes: G01N 30/38 Flow patterns
• CPC Class Codes: G01N 30/6095 Micromachined or nanomachined, e.g. micro- or nanosize
• CPC Class Codes: G01N 30/74 : Optical detectors measurement of intensity, velocity, spectral content, polarisation, or phase of infrared, visible or ultraviolet light G01J
• CPC Class Codes: G01N 35/085 : Flow Injection Analysis
• CPC Class Codes: Y10T 436/117497 : with a continuously flowing sample or carrier stream
• CPC Class Codes: Y10T 436/118339 : with formation of a segmented stream
• CPC Class Codes: Y10T 436/25375 : Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

A reference T-sensor system is provided for detecting the presence and/or measuring the concentration of analyte particles in a sample stream. The system includes: a) a laminar flow channel; b) three or more inlets in fluid connection with the laminar flow channel for respectively conducting into the laminar flow channel (1) an indicator stream which may include an indicator substance which indicates the presence of analyte particles by a detectable change in property when contacted with the analyte particles, (2) the sample stream, and (3) a reference stream, which can be a control stream and/or an internal standard stream; c) wherein the laminar flow channel has a depth and/or width sufficiently small to allow laminar flow of the streams and a length sufficient to allow particles of the analyte to diffuse into the indicator stream to form a detection area; and (d) an outlet for conducting the streams out of the laminar flow channel preferably to form a single mixed stream.

Claims

1. A device for detecting the presence or determining the concentration of analyte particles in a sample stream comprising:

(a) a laminar flow channel;

(b) a sample stream within said laminar flow channel;

(c) an indicator stream within said laminar flow channel in laminar flow contact with said sample stream; and

(d) a reference stream within said laminar flow channel in laminar flow contact with said indicator stream.

2. The device of claim 1 wherein said reference stream is selected from the group consisting of control streams, internal standard streams, and calibration streams.

3. The device of claim 1 also comprising detecting means positioned relative to said flow channel such that said detecting means can detect a change in a detectable property in a detection area in at least one of said streams.

4. The device of claim 3 wherein said detecting means comprise components selected from the group consisting of a charge coupled device camera, a diode array detector, a fluorescence detector, and an electrochemical detector.

5. The device of claim 3 wherein said detection area is an area in said indicator stream into which analyte particles have diffused.

6. The device of claim 3 wherein said detection area is an area in said indicator stream into which particles contained within said reference stream have diffused.

7. The device of claim 3 wherein said detection area is an area in said indicator stream into which no particles have diffused.

8. The device of claim 3 comprising at least two detection areas.

9. The device of claim 1 also comprising means for controlling flow of at least one of said streams.

10. The device of claim 9 wherein a single flow control means controls flow of both said sample and said reference streams.

11. The device of claim 10 comprising second flow control means for controlling flow of said indicator stream at a different rate from said sample and reference streams.

12. The device of claim 1 also comprising a second indicator stream within said laminar flow channel.

13. The device of claim 12 wherein said indicator stream and said second indicator stream flow in adjacent laminar flow flanked by said sample stream and said reference stream.

14. A system for detecting the presence or determining the concentration of analyte particles in a sample stream comprising:

(a) a device comprising;

(i) a laminar flow channel;

(ii) at least three inlet means in fluidic connection with said laminar flow channel for respectively conducting into said laminar flow channel (1) an indicator stream, (2) a sample stream, and (3) a reference stream; and

(iii) detection means positioned relative to said flow channel to measure a value for a detectable property in at least one of said streams; and

(b) means for correlating said measured value with standard values for said detectable property in said system.

15. The system of claim 14 wherein said means for correlating comprise calibration curves.

16. The system of claim 15 wherein said means for correlating comprise software embodying said calibration curves.

17. The system of claim 16 wherein said software provides for entry of measured values from said detecting means and correlates said entries with said calibration curves to provide a value for said detectable property in said flow channel.

18. The system of claim 14 wherein said laminar flow channel has a depth between about 20 micrometers and about 1 mm.

19. The system of claim 14 wherein said laminar flow channel has a width between about 100 micrometers and about 400 micrometers.

20. The system of claim 14 wherein said laminar flow channel has a length between about 5 mm and about 50 mm.

21. The system of claim 14 wherein said sample stream comprises a fluid selected from the group consisting of blood, drinking water, organic solvents, urine, and biotechnological process samples.

22. A device for detecting the presence or determining the concentration of analyte particles in a sample stream comprising:

(a) a substrate;

(b) a plurality of laminar flow channels within said substrate;

(c) a plurality of indicator stream inlet channels in fluidic connection with each of said laminar flow channels for conducting an indicator stream into each of the laminar flow channels;

(d) a sample stream inlet channel in fluidic connection with at least one of said laminar flow channels, positioned for conducting a sample stream therein into laminar flow contact with an indicator stream, and

(e) a single detection means positioned relative to said laminar flow channels so as to detect changes in fluids within all of said laminar flow channels.

23. The device of claim 22 also comprising a reference stream inlet in fluidic connection with at least one of said laminar flow channels for conducting a reference stream therein into laminar flow contact with said indicator stream.

24. The device of claim 22 wherein said inlet channels are on different planes within said substrate.

25. The device of claim 22 wherein said inlet channels are on the same plane within said substrate.

26. The device of claim 22 also comprising an indicator manifold channel connected to all of said indicator stream inlet channels.

27. A method for making a system for detecting the presence or determining the concentration of analyte particles in a sample stream comprising:

(a) providing a substrate;

(b) forming a laminar flow channel in said substrate;

(c) forming at least three inlet means in fluidic connection with said laminar flow channel in said substrate;

(d) flowing a reagent stream into an inlet means between two of the other inlet means;

(e) flowing a sample stream into at least one of the other inlet means adjacent the reagent inlet means; and

(f) detecting the presence or determining the concentration of analyte particles in a diffusion zone between the reagent stream and the adjacent sample stream.

28. The method of claim 27 also comprising preparing calibration curves for an analyte in a sample stream to be analyzed in said system.

29. A method for calibrating a device for determining the concentration of analyte particles in a sample stream comprising:

(a) conducting a plurality of calibration streams, each containing a different known concentration of analyte particles, into one or more laminar flow channels of said device;

(b) conducting a plurality of indicator streams, each having the same composition and comprising an indicator substance which indicates the presence of said analyte particles by a change in a detectable property when contacted with particles of said analyte, into said laminar flow channel(s), whereby each of said calibration streams is in laminar flow with at least one of said indicator streams;

(c) conducting a first internal standard stream having a known concentration of analyte particles higher than that of said calibration streams, and a second internal standard stream having a known concentration of analyte particles lower than that of said calibration streams, into laminar flow with at least one of said indicator streams;

(d) measuring a value of a detectable property in said indicator streams indicative of the concentration of analyte particles in adjacent calibration or internal standard streams; and

(e) plotting the measured values for said calibration streams against the average value for said internal standard streams to obtain a calibration curve for said device.

30. A method for detecting the presence or determining the concentration of analyte particles in a sample stream, comprising:

(a) continuously conducting said sample stream into a laminar flow channel;

(b) continuously conducting an indicator stream, said indicator stream comprising an indicator substance which indicates the presence of said analyte particles by a change in a detectable property when contacted with particles of said analyte, into said laminar flow channel, whereby said sample stream and said indicator stream flow in adjacent laminar flow in said channel;

(c) continuously conducting a reference stream, comprising a constant concentration of 0 or greater of reference particles into said laminar flow channel, whereby said reference stream flows in a laminar stream adjacent to said indicator stream;

(d) allowing analyte particles to diffuse into said indicator stream;

(e) allowing reference particles to diffuse into said indicator stream; and

(f) continuously monitoring the presence or determining the concentration of said analyte and reference particles in said indicator stream.

31. The method of claim 30 comprising monitoring flow rate and correcting for fluctuations in flow rate within said laminar flow channel.