Self-encoding sensor with microspheres

• US 7,348,181 B2
• Filed: 04/06/1999
• Issued: 03/25/2008
• Est. Priority Date: 10/06/1997
• Status: Expired due to Fees

• Current Assignee: Trustees of Tufts College
• Original Assignee: Trustees of Tufts College
• Inventors: Walt, David R., Dickinson, Todd A.
• Primary Examiner(s): GABEL, GAILENE
• Application Number:
  US09/287,573
• Publication Number:
  US 20010029049A1
• Time in Patent Office: 3,276 Days
• Field of Search: 422/55, 422/68.1, 422/82.05, 422/82.06, 422/82.07, 422/82.08, 422/82.09, 422/91, 435/7.92, 435/6, 435/808, 435/91.2, 435/7.1, 435/91, 435/287.2, 435/288.4, 436/164, 436/171, 436/172, 436/7.24, 436/536, 436/518, 436/19, 385/12, 385/38, 385/147
• US Class Current: 436/172
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/0045 using optical fibres
• CPC Class Codes: B01J 2219/00466 in a slurry
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00524 in the shape of fiber bundles
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00648 by the use of solid beads
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: C12Q 1/682 Signal amplification
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/7703 using reagent-clad optical fibres or optical waveguides using measurement of total internal reflection or attenuated total reflection G01N21/552; optical fibres or waveguides per se G02B
• CPC Class Codes: G01N 2201/0846 Fibre interface with sample, e.g. for spatial resolution
• CPC Class Codes: G01N 33/0031 comprising two or more sensors, e.g. a sensor array electrochemical electrode arrays G01N27/27
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form
• CPC Class Codes: Y10T 436/101666 Particle count or volume standard or control [e.g., platelet count standards, etc.]
• CPC Class Codes: Y10T 436/109163 Inorganic standards or controls

Abstract

A microsphere-based analytic chemistry system is disclosed in which self-encoding microspheres having distinct characteristic optical response signatures to specific target analytes may be mixed together while the ability is retained to identify the sensor type and location of each sensor in a random dispersion of large numbers of such sensors in a sensor array using an optically interrogatable encoding scheme. An optical fiber bundle sensor is also disclosed in which individual microsphere sensors are disposed in microwells at a distal end of the fiber bundle and are optically coupled to discrete fibers or groups of fibers within the bundle. The identities of the individual sensors in the array are self-encoded by exposing the array to a reference analyte while illuminating the array with excitation light energy. A single sensor array may carry thousands of discrete sensing elements whose combined signal provides for substantial improvements in sensor detection limits, response times and signal-to-noise ratios.

Claims

1. A method of statistically analyzing response signals obtained from a sensor element array to determine the statistical validity of said response signals, said method comprising:

a) providing an array comprising a population of sensor elements at a density of least 20,000 sensor elements per 1 mm², said population of sensor elements comprising a plurality of subpopulations of sensor elements, the plurality of subpopulations of sensor elements comprising a first subpopulation comprising sensor elements having the same first bioactive agent and a second subpopulation comprising sensor elements having the same second bioactive agent;

b) contacting said array with a composition comprising at least a target analyte, thereby producing a response signal at said sensor elements of at least one of said first and second subpopulations;

c) obtaining individual response signals from 5 to 100 separate sensor elements from at least one of said first and second subpopulations; and

d) performing a statistical analysis on said response signals from at least one of said first and second subpopulations, whereby statistical validity of said response signals is determined.

2. The method of claim 1, wherein said sensor elements are beads and said array comprises subpopulations of beads dispersed on a substrate.

3. The method of claim 2 wherein said substrate is a fiber optic bundle.

4. The method of claim 2 further comprising identifying the location of sensor element within each sensor subpopulation within the array.

5. The method according to claim 2, further comprising determining outlying beads and excluding outlying beads from said subpopulation.

6. The method according to claim 2, wherein said substrate is selected from the group consisting of glass and plastic.

7. The method according to claim 1, wherein at least one of said bioactive agents is a nucleic acid.

8. The method according to claim 7, wherein said nucleic acid is 25 nucleobases in length.

9. The method according to claim 7, wherein said nucleic acids comprise nucleic acid pairs, said nucleic acid pairs having the same sequence at all but one nucleotide position.

10. The method according to claim 1, wherein at least one of said bioactive agents is a protein.

11. The method according to claim 1, wherein said statistical analysis comprises calculating the mean of at least said response signals from said first of said plurality of subpopulations.

12. The method according to claim 1, wherein said statistical analysis comprises calculating the standard deviation of at least said response signals from said first of said plurality of subpopulations.

13. The method according to claim 1, further comprising evaluating the statistical validity of said response signals.

14. The method according to claim 1, further comprising performing a second statistical analysis on said response signals.

15. The method according to claim 14, wherein said second statistical analysis comprises evaluating said measurements using confidence intervals.

16. The method according claim 14, wherein said second statistical analysis comprises using said response signals to perform hypothesis testing.

17. The method according to claim 1, further comprising comparing said statistical analysis of response signals obtained from at least two subpopulations.

18. The method according to claim 17, herein said statistical analysis comprises performing a cluster analysis of response signals from each of said subpopulations.

19. The method according to claim 1, wherein individual response signals from 5 to 20 separate response elements are obtained.

20. The method according to claim 1, wherein individual response signals from roughly 10 separate response elements are obtained.

21. The method according to claim 1, wherein individual response signals from 10 separate response elements are obtained.

22. The method according to claim 1, wherein said sensor elements are attached to a planar substrate.

23. The method according to claim 22, wherein said composition does not flow-through said substrate.

24. The method according to claim 22, wherein said sensor elements are arranged in a regular pattern addressable in the X-Y coordinate plane.

25. The method according to claim 22, wherein said sensor elements are randomly distributed on said planar substrate.

26. The method according to claim 22, wherein said planar substrate is selected from the group consisting of glass and plastic.

27. The method according to claim 22, wherein said planar substrate is approximately the dimensions of a microscope slide cover slip.

28. The method according to claim 1, wherein said target analyte is present in solution.

29. The method according to claim 28, wherein said solution is applied directly to said sensor elements.

30. The method according to claim 29, wherein said sensor elements are immersed in said solution.

31. The method according to claim 30, wherein said sensor elements are immersed in said solution by dipping the sensor elements in said solution.