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

• US 6,171,865 B1
• Filed: 08/04/1999
• Issued: 01/09/2001
• Est. Priority Date: 03/29/1996
• Status: Expired due to Term

• Current Assignee: University of Washington
• Original Assignee: University of Washington
• Inventors: Weigl, Bernhard H., Holl, Mark R., Kenny, Margaret, Zebert, Diane, Wu, Caicai
• Primary Examiner(s): WALLENHORST, MAUREEN
• Application Number:
  US09/366,821
• Time in Patent Office: 524 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 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 inlet means 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 the analyte particles, (2) the sample stream, and (3) a reference stream, which can be a control stream and/or internal standard stream; and, c) wherein the laminar flow channel has a depth and/or width sufficiently small to allow laminar flow of said streams and a length sufficient to allow particles of the analyte to diffuse into the indicator stream to form a detection area. Branching channels may be provided as outlet means for conducting the streams out of the laminar flow channel. Also provided are methods of using the device to improve accuracy and increase convenience of measuring the concentration of analytes in a sample. Monitoring reference stream(s) in laminar flow with sample stream(s) allows for calibration or for monitoring or correcting for the effects of experimental conditions which might otherwise degrade accuracy. A reference stream which serves as both a control stream and an internal standard stream is convenient and economical. The device of this invention also needs less frequent calibration than previously-known fluid analysis devices because the devices and methods of this invention provide for continuous and simultaneous quality control and internal standardization.

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) at least three inlet means in fluid 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

c) wherein said laminar flow channel has a dimension sufficiently small to allow laminar flow of said streams adjacent to each other and a length sufficient to allow analyte particles to diffuse from at least one stream selected from the group consisting of said sample stream and said reference stream into the indicator stream to form at least one detection area.

2. A device of claim 1 also comprising outlet means comprising branching channels for separation of the streams into one or more channels.

3. The device of claim 2 wherein said outlet means comprise smaller channels for conducting the sample stream than for conducting the indicator stream.

4. A device of claim 1 further comprising detecting means positioned relative to said flow channel such that said detecting means can detect a change in a detectable property in at least one of said streams.

5. A device of claim 4 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.

6. A device of claim 1 further comprising inlet means for conducting at least one additional reference or sample stream in laminar flow contact with said indicator stream.

7. A device of claim 1 further comprising means for dividing said indicator or sample stream into at least two separate streams and conducting said separate streams into said laminar flow channel.

8. A device of claim 1 further comprising a plurality of laminar flow channels in fluid communication with an indicator stream channel and means for conducting portions of an indicator stream from said indicator stream channel into laminar flow with separate sample or reference streams in said laminar flow channels.

9. A device of claim 1 further comprising means for conducting at least two streams, each made up of at least two streams in laminar flow with each other, into parallel laminar flow in said laminar flow channel.

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

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

b) 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) 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;

f) detecting the presence or determining the concentration of said analyte and reference particles in said indicator stream; and

g) separately conducting at least one of said streams out of said laminar flow channel.

11. The method of claim 10, wherein the reference stream contains a concentration of analyte particles which is greater than zero.

12. The method of claim 10, wherein two indicator streams are conducted into said laminar flow channel in adjacent laminar flow to said sample stream.

13. The method of claim 10, wherein two reference streams are conducted into said laminar flow channel in adjacent laminar flow to said indicator stream.

14. A method of claim 10, wherein said reference stream is a control stream.

15. A method of claim 10, wherein said reference stream is an internal standard stream.

16. A method of claim 15, wherein said internal standard stream contains reference particles different from said analyte particles.

17. A method of claim 15, wherein said internal standard stream contains reference particles the same as said analyte particles.

18. A method of claim 10, wherein said reference stream is used both as a control stream and as an internal standard stream.

19. A method of claim 10, wherein said reference stream is used as a calibration stream.

20. A method of claim 10 wherein the detectable property is selected from the group consisting of absorbance, chemiluminescence and fluorescence.

21. A method of claim 10, wherein the indicator substance is immobilized on a particulate substance carried within the indicator stream.

22. The method of claim 10 wherein step (g) comprises conducting the sample stream from said indicator stream.