Capillary electrophoresis detection

• US 5,312,535 A
• Filed: 07/17/1992
• Issued: 05/17/1994
• Est. Priority Date: 07/17/1992
• Status: Expired due to Term

• Current Assignee: Beckman Instruments, Inc. (Danaher Corporation)
• Original Assignee: Beckman Instruments, Inc. (Danaher Corporation)
• Inventors: Waska, Frank L., Johnson, Wayne S., Klein, Gerald L.
• Primary Examiner(s): Niebling, John
• Assistant Examiner(s): DELACROIX MUIRHEI, CYBILLE
• Application Number:
  US07/917,640
• Time in Patent Office: 669 Days
• Field of Search: 356/344, 356/441, 204/299 R
• US Class Current: 204/603
• CPC Class Codes: G01N 27/44721 by optical means

Abstract

Detecting light in a capillary electrophoresis optical system includes input and output windows in the outside surface of a capillary tube. The width of the windows is not substantially greater than the bore of the tube. A fiber optic input is spaced from the capillary tube so that the light is directed to the tube and is matched to the window width. The light output from the tube is matched to be received by a fiber optic output spaced from the capillary tube. The optical light envelop from the fiber input through the tube and to the fiber output is governed by the LaGrange Invariant. The matching is effected in accordance with the Numerical Aperture of the fiber optics.

Claims

1. A capillary electrophoresis system comprising a capillary tube having a bore, an inner surface defining the bore, an outer surface, and a wall thickness of the tube defined between the surfaces, the bore having a width and being for transporting fluid, an input window in an outer surface portion of the tube and an output window in an outer surface portion of the tube, the input window and output window being spaced apart, at least one of the input window or output window defining a window width, and wherein the window width is not substantially greater than the bore width.

2. A system as claimed in claim 1 wherein the width of both the windows are not greater than the bore width.

3. A system as claimed in claim 1 wherein the width of at least one of the windows is equal to the bore width.

4. A system as claimed in claim 3 wherein the width of both the windows is equal to the bore width.

10. A system as claimed in claim 1 including an fiber optic input for directing input light to the input window, the fiber optic input having a Numerical Aperture and wherein the fiber optic input includes a core having an end face spaced a distance from the input window whereby light is transmitted from the fiber optic input end face, the end face having an interface and wherein lines of light from the interface are directed at a converging angle substantially equal to a half angle for the Numerical Aperture, and wherein the converging lines of light are directed towards the input window at a location defining a width substantially equal to the window width.

11. A system as claimed in claim 10 wherein the lines of light at the half angle for a Numerical Aperture of the fiber are directed towards a center of the bore.

12. A system as claimed in claim 10 including a fiber optic output for receiving light from the output window.

13. A system as claimed in claim 1 including an fiber optic output for receiving light from the output window, and the fiber optic output having a Numerical Aperture, and wherein the fiber optic output includes a core having an output end face spaced a distance from the output window, the end face having an interface with a cladding and wherein output lines of light from the output window define a diverging angle related to the Numerical Aperture, the diverging angle of light from the interface of the output window impacting the output fiber, such lines defining a width at the end face at least substantially no wider than the width of the end face of the fiber optic output.

14. A system as claimed in claim 13 wherein the diverging lines of light emanate from a center of the bore.

15. A system as claimed in claim 1 wherein the capillary tube includes a coating and the windows are formed by selective removal of the coating.

19. A system as claimed in claim 10 wherein the capillary tube includes a coating and the windows are formed by selective removal of the coating.

20. A system as claimed in claim 13 wherein the capillary tube includes a coating and the windows are formed by selective removal of the coating.

21. A system as claimed in claim 1 including multiple input windows spaced from each other and multiple output windows, the windows being spaced from each other.

22. A system as claimed in claim 1 wherein the input window is located opposite the output window.

23. A system as claimed in claim 21 wherein respective input windows are related to opposite output windows.

24. A system as claimed in claim 1 wherein the windows have a rectangular cross-section.

5. A capillary electrophoresis system comprising a capillary tube having a bore, the bore having a center, an inner surface defining the bore with a selected diameter, an outer surface, a wall thickness of the tube being defined between the surfaces, the bore being for transporting fluid past a detection path, an input window on an outer surface portion of the bore, the input window having opposite interfaces with the surface thereby defining the input window width, and an output window on an opposite outer surface portion of the tube, the output window having opposite interfaces with the surface thereby defining the output window width, and wherein the input and output window widths are additionally defined by a pair of intersecting straight lines directed from each respective opposite interface of an input window radially through the center of the bore to each respective opposite interface of the output window, and wherein the intersecting lines define a LaGrange Invariant for the detection path.

16. A system as claimed in claim 5 wherein the capillary tube includes a coating and the windows are formed by selective removal of the coating.

6. A capillary electrophoresis system comprising a capillary tube having a bore, an inner surface defining the bore with a selected diameter, an outer surface, a wall thickness of the tube being defined between the surfaces, the bore being for transporting fluid past a detection path, an input window on an outer surface portion of the tube and an output window on an opposite outer surface portion of the tube, wherein the input window and the output window define respective widths, the detection path being defined between the input window through the bore and to the output window, and wherein there is a LaGrange Invariant for the detection path and wherein the input and output windows have a width such that the LaGrange Invariant is substantially consistent through the detection path.

7. A system as claimed in claim 6 wherein the LaGrange Invariant is determined by the substantially maximum angle for receiving a signal in the bore relative to the substantially least signal through the tube wall.

8. A system as claimed in claim 7 wherein the LaGrange Invariant is selected to be the substantially highest value for the detection path.

17. A system as claimed in claim 6 wherein the capillary tube includes a coating and the windows are formed by selective removal of the coating.

9. A capillary electrophoresis system comprising a capillary tube having a bore, the bore having a center, an inner surface defining the bore with a selected diameter, an outer surface, a wall thickness of the tube being defined between the surfaces, the bore being for transporting fluid past a detection path, an input window on an outer surface portion of the tube, the input window having opposite interfaces with the outer surface of the tube thereby defining the input window width, and an output window on an opposite outer surface portion of the bore, the output window having opposite interfaces with the outer surface of the tube thereby defining the output window width, an axis through the center of the bore and a position midway between the interfaces of the input window and midway between the interfaces of the output window, and wherein the input and output window widths are established by H=Yn₁ u₀ wherein Y is substantially half the window widths and substantially half the diameter of the bore, n₁ is the refractive index of the tube, u₀ is the angle between the axis and the line from an interface of the windows through the center, and H is an optical constant for the system.

18. A system as claimed in claim 9 wherein the capillary tube includes a coating and the windows are formed by selective removal of the coating.

25. A capillary electrophoresis system comprising a capillary tube having a bore, an inner surface defining the bore diameter, an outer surface, a wall thickness of the tube being defined between the surfaces, the bore being for transporting fluid past a detection path, an input window on an outer surface portion of the tube and an output window on an opposite outer surface portion of the tube, at least one of the input window and output window defining a width, the width being substantially equal to the bore diameter, means for generating a signal, and wherein the signal directed along the detection path between the input and output windows essentially does not pass through the wall without additionally passing through the bore.

26. A system as claimed in claim 25 including a fiber optic input having a core and cladding for directing input light to the input window, and wherein the fiber optic input includes an end face for the core, the end face being spaced a distance from the input window whereby light related to a Numerical Aperture of the fiber optic input from an interface of the core and cladding defines a substantially straight line adjacent an edge of the input window width and to the center of the bore.

27. A system as claimed in claim 25 including a fiber optic output including a core and cladding for receiving output light from the output window, and wherein the fiber optic output includes an end face for the core, the end face being spaced a distance from the output window whereby output light related to a Numerical Aperture of the fiber optic output directed along a substantially straight line from the center of the bore adjacent an edge of the output window width is directed to a position at least within the interface of the core and cladding.

28. A system as claimed in claim 26 including a fiber optic output including a core and cladding for receiving output light from the output window, and wherein the fiber optic output includes an end face for the core, the end face being spaced a distance from the output window whereby the light from the center of the bore directed adjacent an edge of the output window width is directed to a position at least within the interface of the core and cladding.

29. A capillary electrophoresis optical system comprising a capillary tube having a bore with a center, an inner surface defining the bore with a selected diameter, an outer surface, and a wall thickness of the tube defined between the surfaces, the bore being for transporting fluid past an optical path, an optical input window in an outer surface of the tube and an optical output window in an opposite outer surface portion of the tube, at least one of the input window or output window defining an aperture width, and wherein the window width is not substantially greater than the bore diameter.

30. A system as claimed in claim 29 wherein the width of both the windows are not greater than the bore diameter.

31. A system as claimed in claim 29 wherein the width of at least one of the windows is equal to the bore diameter.

32. A system as claimed in claim 31 wherein the width of both the windows is equal to the bore diameter.

38. A system as claimed in claim 29 including a fiber optic input for directing input light to the input window, the fiber optic input having a Numerical Aperture, and wherein the fiber optic input includes a core having an end face spaced a distance from the input window whereby light is transmitted from the fiber optic input end face, the end face having an interface with a cladding and wherein lines of light from the interface are directed at a converging angle substantially equal to a half angle for the Numerical Aperture, and wherein such converging lines of light are directed towards the input window to impact the input window at a location defining a width substantially equal to the window width.

39. A system as claimed in claim 38 wherein the lines of light at the half angle for a Numerical Aperture of the fiber are directed towards the center of the bore.

40. A system as claimed in claim 29 including a fiber optic output for receiving light from the output window.

41. A system as claimed in claim 29 including a fiber optic output for receiving light from the output window, and the fiber optic output having a Numerical Aperture, and wherein the fiber optic output includes a core having an output end face spaced a distance from the output window, the end face having an interface with a cladding and wherein output lines of light from the output window define an angular diverging angle related to the Numerical Aperture, the diverging angle being such that when lines of light from the interface of the output window impact the output fiber, such lines define a width at least substantially no wider than the width of the end face of the fiber optic output.

42. A system as claimed in claim 41 wherein the diverging lines of light emanate from the center of the bore.

43. A system as claimed in claim 29 wherein the capillary tube includes a coating and the windows are formed by selective removal of the coating.

47. A system as claimed in claim 38 wherein the capillary tube includes a coating and the windows are formed by selective removal of the coating.

48. A system as claimed in claim 41 wherein the capillary tube includes a coating and the windows are formed by selective removal of the coating.

49. A system as claimed in claim 29 including multiple input windows spaced from each other and multiple output windows, the windows spaced from each other.

50. A system as claimed in claim 29 wherein the input window is located radially opposite the output window.

51. A system as claimed in claim 49 wherein respective input windows are related to radially opposite output windows.

52. A system as claimed in claim 29 wherein the windows have a rectangular cross-section.

33. A capillary electrophoresis optical system comprising a capillary tube having a bore, the bore having a center, an inner surface defining the bore with a selected diameter, an outer surface, a wall thickness of the tube being defined between the surface, the bore being for transporting fluid past an optical path, an optical input window on an outer surface portion of the bore, the input window having opposite interfaces with the surface thereby defining the input window width, and an optical output window on an opposite outer surface portion of the tube, the output window having opposite interfaces with the surface thereby defining the output window width, and wherein the input and output window widths are defined by a pair of intersecting straight lines directed from each respective opposite interface of an input window radially through the center of the bore to each respective opposite interface of the output window and the window widths are substantially equal to the bore diameter.

44. A system as claimed in claim 33 wherein the capillary tube includes a coating and the windows are formed by selective removal of the coating.

34. A capillary electrophoresis optical system comprising a capillary tube having a bore, an inner surface defining the bore with a selected diameter, an outer surface, a wall thickness of the tube being defined between the surfaces, the bore being for transporting fluid past an optical path, an optical input window on an outer surface portion of the tube and an optical output window on an opposite outer surface portion of the tube, wherein the input window and the output window define respective widths, the optical path being defined between the input window and output window and wherein there is a LaGrange Invariant for the optical path and wherein the LaGrange Invariant is substantially consistent through the optical path.

35. A system as claimed in claim 34 wherein the LaGrange Invariant is determined by the substantially maximum angle for receiving light in the bore relative to the substantially least light through the tube wall.

36. A system as claimed in claim 35 wherein the LaGrange Invariant is selected to be substantially the highest value for the optical path.

45. A system as claimed in claim 34 wherein the capillary tube includes a coating and the windows are formed by selective removal of the coating.

37. A capillary electrophoresis optical system comprising a capillary tube having a bore, the bore having a center, an inner surface defining the bore with a selected diameter, an outer surface, a wall thickness of the tube being defined between the surfaces, the bore being for transporting fluid past an optical path, an optical input window on an outer surface portion of the tube, the input window having opposite interfaces with the outer surface of the tube thereby defining the input window width, and an optical output window on an opposite outer surface portion of the bore, the output window having opposite interfaces with the outer surface of the tube thereby defining the output window width, an optical axis through the center of the bore and a position midway between the interfaces of input window and midway between the interfaces of the output window, and wherein the input and output window widths are established by H=Yn₁ u₀ wherein Y is substantially half the window widths and substantially half the diameter of the bore, n₁ is the refractive index of the tube, u₀ is the angle between the optical axis and the line from an interface of the windows through the center, and H is an optical constant for the system.

46. A system as claimed in claim 37 wherein the capillary tube includes a coating and the windows are formed by selective removal of the coating.

53. A capillary electrophoresis optical system comprising a capillary tube having a bore with a center, an inner surface defining the bore with a selected diameter, an outer surface, a wall thickness of the tube being defined between the surfaces, the bore being for transporting fluid past an optical path, an optical input to the tube and an optical output to the tube, and wherein the bore diameter defines a LaGrange Invariant, and wherein the optical path between the input and output conforms to the LaGrange Invariant.

54. A system as claimed in claim 53 including a fiber optic input for directing input light to the optical input, and wherein the fiber optic input defines a Numerical Aperture and wherein the Numerical Aperture is related to the LaGrange Invariant of the tube.

55. A system as claimed in claim 53 including an fiber optic output for receiving output light from the optical output, and wherein the fiber optic output defines a Numerical Aperture, and wherein the Numerical Aperture is related to the LaGrange Invariant of the tube.

56. A system as claimed in claim 54 including a fiber optic output for receiving output light from the output window, and defines a Numerical Aperture, and wherein the Numerical Aperture is matched in relation to the LaGrange Invariant of the tube.

57. A capillary electrophoresis optical system comprising a capillary tube having a bore with a center, an inner surface defining the bore with a selected diameter, an outer surface, and a wall thickness of the tube defined between the surfaces, the bore being for transporting fluid past an optical path, a fiber optic input for directing input light to the capillary, and wherein the fiber optic input includes a core having an end face spaced a distance from the input window whereby light is transmitted from the fiber optic input end face, the end face having an interface with a cladding and wherein light from the interface is directed at a converging angle related to a Numerical Aperture of the fiber input, a fiber optic output for receiving light from the capillary, and wherein the fiber optic output includes a core having an output end face spaced a distance from the capillary, the fiber optic output including a core having an interface with a cladding and wherein light is directed towards the interface, and wherein the light direction from the interface define a LaGrange Invariant for the optical path.

58. A system as claimed in claim 57 wherein the LaGrange Invariant is defined by a Numerical Aperture of the fiber optic input and fiber optic output.