Rapid detection of species bands in a separation conduit

• US 20020097398A1
• Filed: 10/12/2001
• Published: 07/25/2002
• Est. Priority Date: 07/16/1996
• Status: Active Grant

• Current Assignee: Caliper Life Sciences Incorporated (Revvity, Inc.)
• Original Assignee: Caliper Holdings Incorporated
• Inventors: Parce, J. Wallace
• Granted Patent: US 6,590,653 B2
• Time in Patent Office: Days
• US Class Current: 356/344
• CPC Class Codes: G01N 27/44721 by optical means
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: Y10S 366/02 Micromixers: segmented laminar flow with boundary mixing orthogonal to the direction of fluid propagation with or without geometry influences from the pathway

Abstract

The present invention provides a microfluidic system for fast, accurate and low cost electrophoretic analysis of materials in the fields of chemistry, biochemistry, biotechnology, molecular biology and numerous other fields. Light from periodically spaced regions along a channel in the microfluidic system are received by a photodetector. The intensity of light received by the photodetector is modulated by the movement of species bands through the channel under electrophoretic forces. By Fourier analysis, the velocity of each species band is determined and the identification of the species is made by its electrophoretic mobility in the channel.

Claims

1. A microfluidic system for high-speed electrophoretic analysis of subject materials, said system comprising

a channel in a substrate, said channel holding said subject materials in solution in an electric field so that said materials move through said channel and separate into species bands according to species;

a light source exciting fluorescent light in said bands;

a photoreceptor arranged to receive said fluorescent light from said bands;

means for masking said channel so that said photoreceptor can receive said fluorescent light only at periodically spaced regions along said channel; and

an unit connected to analyze frequencies of light intensity received by said photoreceptor so that velocities of said bands along said channel can be determined for analysis of said materials.

2. The microfluidic system of claim 1 wherein said masking means comprises a mask interposed between said light source and said channel such that said light source illuminates said channel only at said periodically spaced regions.

3. The microfluidic system of claim 1 wherein said masking means comprises a mask interposed between said channel and said photoreceptor such that said photoreceptor can receive light from said channel only at said periodically spaced regions.

4. The microfluidic system of claim 3 wherein said channel is disposed in a transparent substrate and said masking means comprises opaque material defining a plurality of transparent regions at said periodically spaced regions along said channel.

5. The microfluidic system of claim 1 wherein said periodically spaced regions are spaced apart greater than a width of a band in said channel.

6. The microfluidic system of claim 5 wherein said periodically spaced regions are approximately two times a width of a band in said channel.

7. The microfluidic system of claim 1 wherein said channel has a cross-sectional area less than 1000 μ

m².

8. The microfluidic system of claim 7 wherein said channel is approximately 10μ

deep by 60μ

wide.

9. The microfluidic system of claim 1 wherein said light source comprises an element selected from the group consisting of a lamp, a laser and a light-emitting diode.

10. The microfluidic system of claim 1 wherein said light receptor comprises an element selected from the group consisting of a photomultiplier tube, a photodiode, and a CCD array.

11. The microfluidic system of claim 1 further comprising a filter arranged with respect to said light receptor to block light from said light source.

12. The microfluidic system of claim 1 further comprising a lens arranged between said channel and said photoreceptor, said lens focusing fluorescent light from said channel onto said photoreceptor.

14. The microfluidic system of claim 13 wherein said species bands absorb light from said light source, said frequencies of light intensity created by the absence of light toward said photoreceptor from said species bands moving through said channel.

15. The microfluidic system of claim 13 wherein said species bands absorb light from said light source and emit fluorescent light, said frequencies of light intensity created by the presence of fluorescent light toward said photoreceptor from said species bands moving through said channel.

16. The microfluidic system of claim 13 wherein said masking means comprises a mask interposed between said light source and said channel such that said light source illuminates said channel only at said periodically spaced regions.

17. The microfluidic system of claim 13 wherein said masking means comprises a mask interposed between said channel and said photoreceptor such that said photoreceptor can receive light from said channel only at said periodically spaced regions.

18. The microfluidic system of claim 17 wherein said channel is disposed in a transparent substrate and said masking means comprises opaque material defining a plurality of transparent regions at said periodically spaced regions along said channel.

19. The microfluidic system of claim 13 wherein said periodically spaced regions are spaced apart greater than a width of a band in said channel.

20. The microfluidic system of claim 19 wherein said periodically spaced regions are approximately two times a width of a band in said channel.

13. A microfluidic system for high-speed electrophoretic analysis of subject materials, said system comprising

a channel in a substrate, said channel holding said subject materials in solution in an electric field so that said materials move through said channel and separate into species bands according to species;

a light source arranged to direct light toward said channel and said bands;

a photoreceptor arranged to receive light from said channel;

means for masking said channel so that said photoreceptor can receive light at periodically spaced regions along said channel only; and

an unit connected to analyze frequencies of light intensity received by said photoreceptor so that velocities of said bands along said channel can be determined for analysis of said materials.

21. A microfluidic system for high-speed electrophoretic analysis of subject materials, said system comprising

a channel in a substrate, said channel holding said subject materials in solution in an electric field so that said materials move through said channel and separate into species bands according to species;

a light source exciting fluorescent light in said bands;

a photoreceptor arranged to receive said fluorescent light from said bands;

means for masking said channel so that said photoreceptor can receive said fluorescent light only at periodically spaced regions along said channel; and

an unit connected to analyze frequencies of light intensity received by said photoreceptor so that velocities of said bands along said channel can be determined for analysis of said materials.

22. A microfluidic system for high-speed electrophoretic analysis of subject materials, said system comprising

a channel in a substrate, said channel holding said subject materials in solution in an electric field so that said materials move through said channel and separate into species bands according to species;

a light source arranged to direct light toward said channel at periodically spaced regions along said channel only;

a photoreceptor arranged to receive light from said channel; and

an unit connected to analyze frequencies of light intensity received by said photoreceptor so that velocities of said bands along said channel can be determined for analysis of said materials.

23. The microfluidic system of claim 22 wherein said light source comprises a source of coherent light and a pair of slits located between said coherent light source and said channel.

24. The microfluidic system of claim 22 wherein said species bands fluoresces upon exposure to light at said periodically spaced regions along said channel.

25. The microfluidic system of claim 22 wherein said species bands absorbs said light at said periodically spaced regions along said channel.

28. The method of claim 27 wherein said light directing step comprises disposing a mask between a light source and said channel so that only said periodically spaced regions along said channel are illuminated by said light source.

29. The microfluidic system of claim 28 wherein said species bands fluoresce upon exposure to light at said periodically spaced regions along said channel.

30. The microfluidic system of claim 28 wherein said species bands absorb light at said periodically spaced regions along said channel.

31. The method of claim 27 wherein said light directing step comprises disposing a plurality of slits between a coherent light source and said channel so that only said periodically spaced regions along said channel are illuminated by interference patterns of light from said light source through said plurality of slits.

32. The microfluidic system of claim 31 wherein said species bands fluoresce upon exposure to light at said periodically spaced regions along said channel.

33. The microfluidic system of claim 31 wherein said species bands absorb said light at said periodically spaced regions along said channel.

34. The method of claim 27 wherein said light receiving step comprises disposing a mask between said channel and a photoreceptor so that only light from said periodically spaced regions along said channel are received by said photoreceptor.

35. The microfluidic system of claim 34 wherein said species bands fluoresce upon exposure to said directed light.

36. The microfluidic system of claim 34 wherein said species bands absorb said directed light.

26. A microfluidic system for high-speed electrophoretic analysis of subject materials, said system comprising

a channel in a substrate, said channel holding said subject materials in solution in an electric field so that said materials move through said channel and separate into species bands according to species;

a light source arranged to direct light toward said channel;

a photoreceptor arranged to receive light from periodically spaced regions along said channel only; and

an unit connected to analyze frequencies of light intensity received by said photoreceptor so that velocities of said bands along said channel can be determined for analysis of said materials.

27. A method of performing high-speed electrophoretic analysis of subject materials, said method comprising

holding said subject materials in solution in a channel of a microfluidic system;

subjecting said materials to an electric field so that said subject materials move through said channel and separate into species bands;

directing light toward said channel;

receiving light from periodically spaced regions along said channel simultaneously; and

analyzing frequencies of light intensity of said received light so that velocities of said bands along said channel can be determined for analysis of said materials.