Fourier detection of species migrating in a microchannel

• US 5,852,495 A
• Filed: 09/30/1997
• Issued: 12/22/1998
• Est. Priority Date: 07/16/1996
• Status: Expired due to Term

• Current Assignee: Caliper Life Sciences Incorporated (Revvity, Inc.)
• Original Assignee: Caliper Holdings Incorporated
• Inventors: Parce, J. Wallace
• Primary Examiner(s): Font, Frank G.
• Assistant Examiner(s): STAFIRA, MICHAEL PATRICK
• Application Number:
  US08/941,679
• Time in Patent Office: 448 Days
• Field of Search: 356/344, 356/318, 204/299 R, 204/182.8, 204/180.1, 250/461.2, 210/198.2, 210/198.3, 210/658, 210/656, 73/61.53, 73/23.39, 73/61.48
• 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 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 at periodically spaced regions along said channel simultaneously;

receiving light from said channel; 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.

2. The method of claim 1 wherein said directing step comprises disposing a pair of slits between a source of coherent light and said channel.

3. The method of claim 1 wherein said species bands fluoresces upon exposure to light at said periodically spaced regions along said channel.

4. The method of claim 3 further comprising the step of labeling each of said species with a fluorescent label material.

5. The method of claim 4 wherein said fluorescent label material comprises a fluorescent intercalating agent.

6. The method of claim 5 wherein said species comprises a polynucleotide and wherein said fluorescent intercalating agent comprises ethidium bromide.

7. The method of claim 5 wherein said species comprises a protein and wherein said fluorescent intercalating agent comprises fluorescein isothiocyanate.

8. The method system of claim 3 wherein each of said species bands fluoresces off and on with a characteristic frequency, wherein said characteristic frequency corresponds to a velocity of said species band along said channel.

9. The method of claim 1 wherein said species bands absorbs said light at said periodically spaced regions along said channel.

10. The method of claim 1 further comprising the step of focusing said light onto said channel.

11. The method of claim 1 further comprising the step of focusing said light from said channel.

12. The method of claim 1 further comprising the step of blocking said light.

13. The method of claim 1 further comprising the step of converting said light from said channel into a plurality of electrical signals.

14. The method of claim 13 further comprising the step of using Fourier analysis to decompose each of said electrical signals into a plurality of electrical signal component frequencies, wherein each of said electrical signal component frequencies comprises a corresponding frequency of said light intensity received by said photoreceptor.

15. The method of claim 14 further comprising the step of automatically relating each of said electrical signal component frequencies to a distinct species of said subject materials.

16. The method of claim 1 further comprising the step of enclosing and fluidly sealing said channel in said substrate.

17. The method of claim 1 further comprising the steps of sealing a first end of said channel and forming a first reservoir at said first end, and sealing a second end of said channel and forming a second reservoir at said second end.

18. The method of claim 17 further comprising the step of simultaneously applying a first selectable voltage level to said first reservoir and a second selectable voltage level to said second reservoir, thereby generating said electric field.

19. A detection apparatus for detecting separated species bands in a microscale channel, comprising:

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

a photoreceptor arranged to receive light from said channel; and

a processor unit connected to said photoreceptor 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 species bands.

20. The detection apparatus of claim 19 wherein said light source comprises a source of coherent light and a pair of slits between said coherent light source and said channel.

21. The detection apparatus of claim 19 wherein said species bands each comprise material which fluoresces upon exposure to light from said light source.

22. The detection apparatus of claim 19 wherein said species bands each comprise material which absorbs light from said light source.

23. The detection apparatus of claim 21 wherein said material comprises a fluorescent label material.

24. The detection apparatus of claim 23 wherein said fluorescent label material comprises a fluorescent intercalating agent.

25. The detection apparatus of claim 24 wherein said species comprises a polynucleotide and wherein said fluorescent intercalating agent comprises ethidium bromide.

26. The detection apparatus of claim 24 wherein said species comprises a protein and wherein said fluorescent intercalating agent comprises fluorescein isothiocyanate.