Analyte detection system

• US 7,300,800 B2
• Filed: 05/13/2005
• Issued: 11/27/2007
• Est. Priority Date: 11/14/2001
• Status: Active Grant

• Current Assignee: Beckman Coulter Incorporated (Danaher Corporation)
• Original Assignee: Beckman Coulter Incorporated (Danaher Corporation)
• Inventors: Pentoney, Stephen L. Jr., Bell, Michael L., Lin, Yuan, Michael, Josephine M., Tsay, Tsong-Tseh
• Primary Examiner(s): Warden; Jill
• Assistant Examiner(s): TURK, NEIL N
• Application Number:
  US11/128,523
• Publication Number:
  US 20050208573A1
• Time in Patent Office: 928 Days
• Field of Search: 436/172, 422/52
• US Class Current: 436/172
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 2015/1438 Using two lasers in succession
• CPC Class Codes: G01N 33/542 with steric inhibition or signal modification, e.g. fluorescent quenching
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/587 Nanoparticles
• CPC Class Codes: Y10T 436/13 Tracers or tags

Abstract

An analyte detection system utilizing a combination of fluorescent labels for labeling particles and an analyte specific fluorescent analyte detection dye. The particles contain a combination of fluorescent labels for coding the particles and an analyte specific fluorescent dye. The particles can be used to identify and quantify analytes in an analytical sample by reaction of the analytical sample with the particles. An analytical device can identify the particles according to the combination of fluorescent labels. The device can then correlate the identified particle with the analyte specific fluorescent analyte detection dye. Multiple subpopulations of particles can be used to identify and quantify multi-analytes in a single analytical sample. Near infrared (NIR) fluorescent labels useful in the detection system are also provided.

Claims

1. A particle comprising:

two or more than two fluorescent labels embedded within the particle in a combination of relative amounts, the fluorescent labels being capable of being excited by light of a same excitation wavelength and capable of emitting lights at maximum wavelengths greater than about 750 nm, distinguishable from each other, respectively.

2. A particle according to claim 1 wherein the fluorescent labels are both cyanine dyes.

3. A particle according to claim 1 wherein the excitation wavelength of the fluorescent labels is greater than 750 nm.

4. A particle according to claim 3 wherein the excitation wavelength is about 780 nm.

5. A particle according to claim 1 wherein the magnitude of the emitting lights of the fluorescent labels corresponds to the relative amounts of fluorescent labels in the particle.

6. A particle according to claim 1 wherein the maximum wavelengths of the emitting lights differ from one another by at least about 30 nm.

7. A particle according to claim 1 wherein the fluorescent labels do not significantly interact through energy transfer.

14. A method for detecting an analyte on a particle comprising:

a) moving one or more than one particle according to claim 1 through an examination zone, each particle further comprising a fluorescent analyte detection dye capable of being excited by light at a first excitation wavelength and capable of emitting light at a maximum wavelength when excited;

b) directing an exciting light of a first wavelength at each particle in the examination zone;

c) directing an exciting light of a second wavelength at each particle in the examination zone, wherein

d) detecting the emitted light of the fluorescent analyte detection dye and the emitted light of the first and second fluorescent labels; and

e) conelating the detected emitted lights with the particle under analysis.

15. A method according to claim 14 wherein the wavelengths of the first and second exciting lights differ by at least 100 nm.

16. A method according to claim 14 wherein the wavelengths of the second exciting light is greater than about 750 nm.

17. A method according to claim 14 wherein the maximum wavelength of the emitted light of the fluorescent analyte detection dye differs from the maximum wavelengths of the emitted lights of each of the fluorescent labels by at least 100 nm.

18. A method according to claim 14 comprising more than one particle, each particle having a different fluorescent analyte detection dye, and two or more than two fluorescent labels in a combination of relative amounts, wherein the combination of fluorescent labels in each particle is different.

19. A method according to claim 14 comprising moving two or more than two particles through an examination zone, each particle having a different size, the method further comprising:

f) directing an exciting light at each particle in the examination zone to generate a scattered light;

g) detecting the scattered light; and

h) correlating the detected scattered light with the emitted lights and the particle under analysis.

20. A method according to claim 14 comprising moving two or more than two particles through an examination zone, the method further comprising:

f) directing an exciting light of a third wavelength at each particle in the examination zone;

g) detecting the emitted light from the third exciting light; and

h) conelating the detected emitted lights with the particle under analysis.

21. A method according to claim 20, each particle including a different size, the method further comprising:

i) directing an exciting light at each particle in the examination zone to generate a scattered light; and

j) detecting the scattered light; and

k) correlating the detected scattered light with the emitted lights and the particle under analysis.

8. An analyte detection system comprising:

a) one or more than one particle, each particle comprising two or more than two fluorescent labels in a combination of relative amounts, wherein each fluorescent label is capable of being excited by light of a same excitation wavelength and capable of emitting light when excited at maximum wavelengths greater than about 750 nm, distinguishable from each other, respectively;

b) means for exciting the fluorescent labels;

c) means for detecting the emitted lights; and

d) means for correlating the detected emitted lights with a particular particle under analysis.

9. An analyte detection system according to claim 8 comprising more than one particle wherein the combination of relative amounts of fluorescent label in each particle is different.

10. An analyte detection system according to claim 8 wherein the means for exciting the fluorescent labels is a laser.

11. An analyte detection system according to claim 8 comprising more than one particle wherein the particles are of different size and including means for illuminating the particles to generate scattered lights, means for detecting the scattered lights, and means for correlating the detected emitted lights and the scattered lights with the particle under analysis.

12. An assay system comprising:

a) a particle having

(i) two or more than two fluorescent labels, each fluorescent label being capable of being excited by light of a same second excitation wavelength and capable of emitting light when excited at maximum wavelengths greater than about 750 nm, distinguishable from each other, respectivley; and

(ii) a receptor;

b) an analyte; and

c) a fluorescent analyte detection dye, wherein the fluorescent analyte detection dye has an emitting light that differs from the wavelengths of emitted lights of each of the fluorescent labels by at least 100 nm.

13. An assay system according to claim 12 further comprising a second receptor, wherein the receptor, the analyte and the second receptor form a fluorescent complex on the particle.