System, method, and product for providing a wavelength tunable excitation beam

• US 20050001176A1
• Filed: 05/13/2004
• Published: 01/06/2005
• Est. Priority Date: 05/13/2003
• Status: Active Grant

• Current Assignee: Affymetrix, Inc. (Thermo Fisher Scientific Incorporated)
• Original Assignee: Affymetrix, Inc. (Thermo Fisher Scientific Incorporated)
• Inventors: Weiner, Nathan K., Loney, Gregory C.
• Granted Patent: US 7,148,492 B2
• Time in Patent Office: Days
• US Class Current: 250/458.100
• CPC Class Codes: G01N 2021/6419 Excitation at two or more wavelengths
• CPC Class Codes: G01N 2021/6471 Special filters, filter wheel
• 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 33/58 involving labelled substances G01N33/53 takes precedence

Abstract

A method for scanning a probe array is described. The method comprises tuning an excitation light to a plurality of wavelengths each within an excitation range of a fluorescent label associated with one or more target molecules; and directing the tuned excitation light of each wavelength at the probe array.

Claims

1. A system for scanning biological probe arrays, comprising:

a biological probe array that comprises a plurality of probes disposed thereon, wherein one or more first probes is associated with a first label and one or more second probes is associated with a second label; and

a source to provide light of a first wavelength and a second wavelength, wherein the first wavelength is within an excitation range of the first label and the second wavelength is within an excitation range of the second label.

2. The system of claim 1, wherein:

at least one of the first probes hybridizes a first target molecule associated with the first label, and at least one of the second probes hybridizes a second target molecule associated with the second label.

3. The system of claim 2, wherein:

the first and second target molecules comprises one or more of the group consisting of DNA, RNA, PNA and Protein molecules.

5. The system of claim 1, wherein:

the first and second labels label emit light in response to the first and second wavelengths.

6. The system of claim 5, further comprising:

a detector to detect the emitted light.

29. The system of claim 1, wherein:

at least one of the target molecules hybridizes a biological probe disposed on the probe array.

7. A method for scanning a probe array, comprising:

tuning an excitation light to a plurality of wavelengths each within an excitation range of a fluorescent label associated with one or more target molecules; and

directing the tuned excitation light of each wavelength at the probe array.

8. The method of claim 7, wherein:

at least one of the target molecules hybridizes a biological probe disposed on the probe array.

9. The method of claim 7, further comprising:

detecting each a second wavelength of light, wherein the second wavelength is emitted in response to the first wavelength;

10. The method of claim 9, wherein:

the first wavelength is different that the second wavelength.

11. The method of claim 9, further comprising:

generating an image based, at least in part, upon each second wavelength.

12. The method of claim 7, wherein:

the step of tuning includes tuning a source.

13. The method of claim 7, wherein:

the step of tuning includes adjusting a distance associated with an internal path length in a laser.

14. The method of claim 7, wherein:

the step of tuning includes translating a wavelength tuning element in a normal plane to the excitation light.

15. The method of claim 14, wherein:

the wavelength tuning element is a wedge etalon or a grating.

16. A method for providing multiple wavelengths of light to a probe array, comprising:

tuning an excitation light to a first wavelength, wherein the first wavelength is within an excitation range of a first label associated with one or more first target molecules;

scanning the probe array with the first wavelength of light;

detecting a second wavelength of light, wherein the second wavelength is emitted in response to the first wavelength;

tuning the excitation light to a third wavelength of light, wherein the third wavelength is within an excitation range of a second fluorescent label associated with one or more second target molecules;

scanning the probe array with the second wavelength of light; and

detecting a fourth wavelength of light, wherein the fourth wavelength is emitted in response to the second wavelength.

17. The method of claim 16, wherein:

at least one of the first and second target molecules each hybridizes a biological probe disposed on the probe array.

18. A method for providing multiple wavelengths of light to a probe array, comprising:

a) tuning an excitation light to a first wavelength, wherein the first wavelength is within an excitation range of a first fluorescent label associated with a first biological probe;

b) scanning a pixel position with the first wavelength of light;

c) detecting a second wavelength of light, wherein the second wavelength is emitted in response to the first wavelength;

d) tuning the excitation light to a third wavelength of light, wherein the third wavelength is within an excitation range of a second fluorescent label associated with a second biological probe;

e) scanning the pixel position with the second wavelength of light;

f) detecting a fourth wavelength of light, wherein the fourth wavelength is emitted in response to the second wavelength; and

g) repeating steps a) through f) until each of a plurality of pixel positions associated with the probe array has been scanned.

19. A method for providing multiple wavelengths of light to a probe array, comprising:

a) tuning an excitation light to a first wavelength, wherein the first wavelength is within an excitation range of a first fluorescent label associated with a first biological probe;

b) scanning a line of pixel positions with the first wavelength of light;

c) detecting a second wavelength of light, wherein the second wavelength is emitted in response to the first wavelength;

d) tuning the excitation light to a third wavelength of light, wherein the third wavelength is within an excitation range of a second fluorescent label associated with a second biological probe;

e) scanning the line of pixel positions with the second wavelength of light;

f) detecting a fourth wavelength of light, wherein the fourth wavelength is emitted in response to the second wavelength; and

g) repeating steps a) through f) until each of a plurality of pixel positions associated with the probe array has been scanned.

20. A system for providing multiple wavelengths of light to a probe array, comprising:

a source to alternately tune an excitation light to a first wavelength within an excitation range of a first fluorescent label associated with a first biological probe, and a second wavelength within an excitation range of a second fluorescent label associated with a second biological probe;

scanner optics to direct the first wavelength and second wavelength of light at a probe array; and

a detector to detect a third wavelength of light emitted in response to the first wavelength and a fourth wavelength of light emitted in response to the second wavelength.

21. The system of claim 20, wherein:

the source holds at the wavelength of the tuned excitation light for a duration of time prior to alternation, wherein the duration includes time to direct the tuned excitation light at a pixel position associated with the probe array.

22. The system of claim 20, wherein:

the source holds at the wavelength of the tuned excitation light for a duration of time prior to alternation, wherein the duration includes time to direct the tuned excitation light at a line of one or more pixel positions associated with the probe array.

23. The system of claim 20, wherein:

the source holds at the wavelength of the tuned excitation light for a duration of time prior to alternation, wherein the duration includes time to direct the tuned excitation light at all pixel positions associated with the probe array.

24. A method for providing multiple wavelengths of light to a probe array, comprising:

tuning an excitation light to a first wavelength, wherein the first wavelength is within an excitation range of a first fluorescent label associated with a first biological probe;

scanning a probe array with the first wavelength of light;

detecting a second wavelength of light, wherein the second wavelength is emitted in response to the first wavelength;

tuning the excitation light to a third wavelength of light, wherein the third wavelength is within an excitation range of a second fluorescent label associated with a second biological probe;

scanning the probe array with the second wavelength of light;

detecting a fourth wavelength of light, wherein the fourth wavelength is emitted in response to the second wavelength; and

generating an image based, at least in part, upon the detected third and fourth wavelengths.

25. A system for providing multiple wavelengths of light to a probe array, comprising:

a source to tune an excitation light to a first wavelength, and second wavelength wherein the first wavelength is within an excitation range of a first fluorescent label associated with a first biological probe, and the second wavelength is within an excitation range of a second fluorescent label associated with a second biological probe;

scanner optics to direct the first wavelength and second wavelengths of light at a probe array;

a detector to detect a third wavelength of light emitted in response to the first wavelength and a fourth wavelength of light emitted in response to the second wavelength; and

a computer having executable code stored thereon, wherein the executable code is enabled to generate an image based, at least in part, upon the detected third and fourth wavelengths.

26. A system for providing multiple wavelengths of light to a probe array, comprising:

a wavelength tuning element to tune an excitation light to a first wavelength, and second wavelength wherein the first wavelength is within an excitation range of a first fluorescent label associated with a first biological probe, and the second wavelength is within an excitation range of a second fluorescent label associated with a second biological probe;

scanner optics to direct the first wavelength and second wavelength of light at a probe array; and

a detector to detect a third wavelength of emitted light responsive to the first wavelength and a fourth wavelength of emitted light responsive to the second wavelength.

27. The system of claim 26, wherein:

the wavelength tuning element is a wedge etalon or a grating.

28. A system for providing multiple wavelengths of light to a probe array, comprising:

a means for tuning an excitation light to a plurality of wavelengths each within an excitation range of a fluorescent label associated with one or more target molecules;

a means for directing the tuned excitation light of each wavelength at the probe array; and

a detector to detect each a second wavelength of light, wherein the second wavelength is emitted in response to the first wavelength.

30. A system for directing multiple wavelengths of light to a probe array and detection of emitted light, comprising:

a scanner to tune an excitation light to a plurality of wavelengths each within an excitation range of a fluorescent label associated with one or more target molecules, and detect each a second wavelength of light, wherein the second wavelength is emitted in response to the first wavelength.

31. The system of claim 30, wherein:

at least one of the target molecules hybridizes a biological probe disposed on the probe array.

32. A system for providing multiple wavelengths of light to a probe array, comprising:

a means for successively tuning an excitation light to a first wavelength within an excitation range of a first fluorescent label associated with a first biological probe, and a second wavelength within an excitation range of a second fluorescent label associated with a second biological probe;

a means for directing the first wavelength and second wavelength of light at a probe array; and

a detector to detect a third wavelength of light emitted in response to the first wavelength and a fourth wavelength of light emitted in response to the second wavelength.