DNA detector and DNA detection method

• US 5,667,656 A
• Filed: 05/14/1996
• Issued: 09/16/1997
• Est. Priority Date: 02/28/1991
• Status: Expired due to Fees

• Current Assignee: Hitachi, Ltd.
• Original Assignee: Hitachi, Ltd.
• Inventors: Kambara, Hideki
• Primary Examiner(s): Gorgos, Kathryn L.
• Assistant Examiner(s): WONG, EDNA
• Application Number:
  US08/645,706
• Time in Patent Office: 490 Days
• Field of Search: 204/603, 204/605, 204/612
• US Class Current: 204/603
• CPC Class Codes: G01N 2035/00237 Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: G01N 27/44721 by optical means
• CPC Class Codes: G01N 27/44782 of a plurality of samples

Abstract

An electrophoresis apparatus for detecting samples migrating in migration portions includes a plurality of gel separation portions separate from one another and having respective ends disposed along a straight line, a plurality of migration portions equal in number to the gel separation portions and disposed along a straight line following respective ones of the gel separation portions, a light source for generating light which passes through the migration portions, the light propagating along a straight line extending through all of the migration portions, a plurality of optical fibers having respective first ends and respective second ends, the first ends of the optical fibers being disposed facing respective points where the light from the light source passes through respective ones of the migration portions, the optical fibers being equal in number to or greater in number than the migration portions, and an optical detector optically coupled to the second ends of the optical fibers for receiving light from the migration portions via the optical fibers.

Claims

1. An electrophoresis apparatus for detecting samples migrating in migration portions, the electrophoresis apparatus comprising:

an optical cell;

a plurality of electrophoretic separation portions separate from one another and having respective ends disposed along a straight line inside the optical cell;

means for forming inside the optical cell a plurality of migration portions equal in number to the electrophoretic separation portions and disposed along a straight line following respective ones of the electrophoretic separation portions;

a light source for generating light which passes through the migration portions, the light propagating along a straight line extending through all of the migration portions;

a plurality of optical fibers having respective first ends and respective second ends, the first ends of the optical fibers being disposed facing respective points where the light from the light source passes through respective ones of the migration portions, the optical fibers being equal in number to or greater in number than the migration portions; and

optical detecting means optically coupled to the second ends of the optical fibers for receiving light from the migration portions via the optical fibers.

2. An electrophoresis apparatus according to claim 1, wherein the electrophoretic separation portions include gel capillaries.

3. An electrophoresis apparatus according to claim 1, wherein the electrophoretic separation portions include electrophoretic migration lanes separate from one another and disposed between two transparent substrates.

4. An electrophoresis apparatus according to claim 1, wherein the light from the light source which passes through the migration portions causes samples migrating in the migration portions to emit fluorescence;

and

wherein the optical detecting means receives the fluorescence from the migration portions via the optical fibers.

5. An electrophoresis apparatus according to claim 1, wherein the optical detecting means includes a one-dimensional optical sensor or a two-dimensional optical sensor.

6. An electrophoresis apparatus according to claim 1, wherein the optical detecting means includes a one-dimensional optical sensor including a plurality of photosensitive elements disposed along a line, or a two-dimensional optical sensor including a plurality of photosensitive elements disposed on a plane;

and

wherein each of the second ends of the optical fibers is optically coupled to at least one of the photosensitive elements of the one-dimensional optical sensor or the two-dimensional optical sensor.

7. An electrophoresis apparatus according to claim 1, further comprising means for establishing a sheath flow around samples migrating in the migration portions.

8. An electrophoresis apparatus according to claim 1, further comprising a plurality of light collecting lenses, each of the light collecting lenses being disposed between (1) a respective one of the points where the light from the light source passes through respective ones of the migration portions and (2) at least one of the first ends of the optical fibers.

9. An electrophoresis apparatus according to claim 1, further comprising a plurality of light collecting lenses, each of the light collecting lenses being optically coupled to a respective one of the first ends of the optical fibers.

10. An electrophoresis apparatus according to claim 1, further comprising a plurality of optical bandpass filters, each of the optical bandpass filters being optically coupled to a respective one of the first ends of the optical fibers.

11. An electrophoresis apparatus according to claim 1, further comprising a plurality of sets of optical bandpass filters, each of the sets of optical bandpass filters including at least two optical bandpass filters, each of the sets of optical bandpass filters being disposed between a respective one of the second ends of the optical fibers and the optical detecting means.

12. An electrophoresis apparatus according to claim 1, further comprising means for splitting the light generated by the light source into a plurality of light beams, and directing one of the light beams into the optical cell such that the one light beam propagates along the straight line extending through all of the migration portions.

13. An electrophoresis apparatus for detecting samples migrating along lines extended from ends of electrophoretic separation portions, the electrophoresis apparatus comprising:

a plurality of electrophoretic separation portions separate from one another and having respective ends disposed along a straight line;

a light source for generating light which passes across respective lines extended from the ends of the electrophoretic separation portions, the light propagating along a straight line extending across all of the lines extended from the ends of the electrophoretic separation portions;

a plurality of optical fibers having respective first ends and respective second ends, the first ends of the optical fibers being disposed facing respective points where the light from the light source passes across respective ones of the lines extended from the ends of the electrophoretic separation portions, the optical fibers being equal in number to or greater in number than the electrophoretic separation portions; and

optical detecting means optically coupled to the second ends of the optical fibers for receiving light from the lines extended from the electrophoretic separation portions via the optical fibers.

14. A fluorescent electrophoresis apparatus comprising:

a plurality of electrophoresis lanes having respective ends disposed along a straight line, the electrophoresis lanes each containing a separation medium through which a sample labeled with a fluorophore migrates until the sample labeled with the fluorophore is eluted from the end of the respective electrophoresis lane;

a plurality of migration portions following respective ones of the electrophoresis lanes and being disposed along a straight line, wherein the sample labeled with the fluorophore eluted from the end of each of the electrophoresis lanes migrates through a respective one of the migration portions;

a light source for generating light which passes through the migration portions along a straight line extending through all of the migration portions, thereby irradiating the migration portions at respective irradiated portions disposed along the straight line extending through all of the migration portions;

a plurality of optical fibers having respective first ends and respective second ends, the first ends of the optical fibers being disposed facing respective ones of the irradiated portions of the migration portions so as to receive fluorescence generated in the irradiated portions, wherein the optical fibers transmit the fluorescence received by the first ends of the optical fibers to the second ends of the optical fibers, thereby causing the fluorescence to be emitted from the second ends of the optical fibers; and

optical detecting means optically coupled to the second ends of the optical fibers for detecting the fluorescence emitted from the second ends of the optical fibers, thereby detecting the sample labeled with the fluorophore eluted from the end of each of the electrophoresis lanes outside the electrophoresis lanes.

15. A fluorescent electrophoresis apparatus according to claim 14, wherein the separation medium is a gel;

and

wherein the electrophoresis lanes are formed in respective ones of a plurality of capillaries each containing the gel.

16. A fluorescent electrophoresis apparatus according to claim 14, wherein the separation medium is a gel;

and

wherein the electrophoresis lanes are formed in a slab of the gel disposed between two substrates.

17. A fluorescent electrophoresis apparatus according to claim 14, wherein the fluorescence is emitted from the fluorophore in response to the light from the light source.

18. A fluorescent electrophoresis apparatus according to claim 14, wherein the optical detecting means includes a one-dimensional optical sensor or a two-dimensional optical sensor.

19. A fluorescent electrophoresis apparatus according to claim 14, wherein the optical detecting means includes a one-dimensional optical sensor or a two-dimensional optical sensor;

and

wherein the second ends of the optical fibers are optically coupled to the one-dimensional optical sensor or the two-dimensional optical sensor.

20. A fluorescent electrophoresis apparatus according to claim 14, further comprising means for forming a sheath flow in each of the migration portions.

21. A fluorescent electrophoresis apparatus according to claim 14, further comprising a plurality of light collecting lenses, each of the light collecting lenses being disposed between (1) a respective one of the irradiated portions of the migration portions and (2) at least one of the first ends of the optical fibers.

22. A fluorescent electrophoresis apparatus according to claim 14, further comprising a plurality of light collecting lenses, each of the light collecting lenses being disposed between (1) at least one of the second ends of the optical fibers and (2) the optical detecting means.

23. A fluorescent electrophoresis apparatus according to claim 14, further comprising a plurality of optical filters, each of the optical filters being optically coupled to a respective one of the first ends of the optical fibers.

24. A fluorescent electrophoresis apparatus according to claim 14, further comprising a plurality of optical band pass filters, each of the optical band pass filters being disposed between a respective one of the second ends of the optical fibers and the optical detecting means.

25. A fluorescent electrophoresis apparatus according to claim 14, wherein the optical fibers are equal in number to or greater in number than the electrophoresis lanes.

26. A fluorescent electrophoresis apparatus comprising:

at least one optical cell;

a plurality of electrophoresis lanes having respective ends disposed along a straight line inside the at least one optical cell, the electrophoresis lanes each containing a separation medium through which a sample labeled with a fluorophore migrates until the sample labeled with the fluorophore is eluted from the end of the respective electrophoresis lane;

a plurality of migration portions formed in the at least one optical cell, the migration portions following respective ones of the electrophoresis lanes and being disposed along a straight line in the at least one optical cell, wherein the sample labeled with the fluorophore eluted from the end of each of the electrophoresis lanes migrates through a respective one of the migration portions;

a light source for generating light which passes through the migration portions along a straight line extending through all of the migration portions, thereby irradiating the migration portions at respective irradiated portions disposed along the straight line extending through all of the migration portions;

a plurality of optical fibers having respective first ends and respective second ends, the first ends of the optical fibers being disposed facing respective ones of the irradiated portions of the migration portions so as to receive fluorescence generated in the irradiated portions, wherein the optical fibers transmit the fluorescence received by the first ends of the optical fibers to the second ends of the optical fibers, thereby causing the fluorescence to be emitted from the second ends of the optical fibers; and

optical detecting means optically coupled to the second ends of the optical fibers for detecting the fluorescence emitted from the second ends of the optical fibers, thereby detecting the sample labeled with the fluorophore eluted from the end of each of the electrophoresis lanes outside the electrophoresis lanes.

27. A fluorescent electrophoresis apparatus according to claim 26, wherein the separation medium is a gel;

and

wherein the electrophoresis lanes are formed in respective ones of a plurality of capillaries each containing the gel.

28. A fluorescent electrophoresis apparatus according to claim 26, wherein the separation medium is a gel;

and

wherein the electrophoresis lanes are formed in a slab of the gel disposed between two substrates.

29. A fluorescent electrophoresis apparatus according to claim 26, wherein the fluorescence is emitted from the fluorophore in response to the light from the light source.

30. A fluorescent electrophoresis apparatus according to claim 26, wherein the optical detecting means includes a one-dimensional optical sensor or a two-dimensional optical sensor.

31. A fluorescent electrophoresis apparatus according to claim 26, wherein the optical detecting means includes a one-dimensional optical sensor or a two-dimensional optical sensor;

and

wherein the second ends of the optical fibers are optically coupled to the one-dimensional optical sensor or the two-dimensional optical sensor.

32. A fluorescent electrophoresis apparatus according to claim 26, further comprising means for forming a sheath flow in each of the migration portions.

33. A fluorescent electrophoresis apparatus according to claim 26, further comprising a plurality of light collecting lenses, each of the light collecting lenses being disposed between (1) a respective one of the irradiated portions of the migration portions and (2) at least one of the first ends of the optical fibers.

34. A fluorescent electrophoresis apparatus according to claim 26, further comprising a plurality of light collecting lenses, each of the light collecting lenses being disposed between (1) at least one of the second ends of the optical fibers and (2) the optical detecting means.

35. A fluorescent electrophoresis apparatus according to claim 26, further comprising a plurality of optical filters, each of the optical filters being optically coupled to a respective one of the first ends of the optical fibers.

36. A fluorescent electrophoresis apparatus according to claim 26, further comprising a plurality of optical band pass filters, each of the optical band pass filters being disposed between a respective one of the second ends of the optical fibers and the optical detecting means.

37. A fluorescent electrophoresis apparatus according to claim 26, wherein the optical fibers are equal in number to or greater in number than the electrophoresis lanes.

38. A fluorescent electrophoresis apparatus according to claim 26, further comprising light splitting means for splitting the light generated by the light source into a plurality of light beams;

wherein the at least one optical cell includes a plurality of optical cells; and

wherein each of the light beams enters at least one of the optical cells so as to irradiate the migration portions therein.

39. A fluorescent electrophoresis apparatus according to claim 26, wherein the at least one optical cell includes a plurality of optical cells;

and

wherein the light generated by the light source enters at least two of the optical cells so as to irradiate the migration portions therein.

40. A fluorescent electrophoresis apparatus comprising:

a plurality of electrophoresis lanes having respective ends disposed along a straight line, the electrophoresis lanes each containing a separation medium through which a sample labeled with a fluorophore migrates until the sample labeled with the fluorophore is eluted from the end of the respective electrophoresis lane;

a plurality of migration portions formed along respective lines extended from the ends of the electrophoresis lanes, the migration portions being disposed along a straight line, wherein the sample labeled with the fluorophore eluted from the end of each of the electrophoresis lanes migrates through a respective one of the migration portions;

a light source for generating light which passes through the migration portions along a straight line extending through all of the migration portions, thereby simultaneously irradiating all of the migration portions at respective irradiated portions disposed along the straight line extending through all of the migration portions;

a plurality of optical fibers having respective first ends and respective second ends, the first ends of the optical fibers being disposed facing respective ones of the irradiated portions of the migration portions so as to receive fluorescence generated in the irradiated portions, wherein the optical fibers transmit the fluorescence received by the first ends of the optical fibers to the second ends of the optical fibers, thereby causing the fluorescence to be emitted from the second ends of the optical fibers; and

optical detecting means optically coupled to the second ends of the optical fibers for detecting the fluorescence emitted from the second ends of the optical fibers, thereby detecting the sample labeled with the fluorophore eluted from the end of each of the electrophoresis lanes outside the electrophoresis lanes.

41. A fluorescent electrophoresis apparatus according to claim 40, wherein the optical fibers are equal in number to or greater in number than the electrophoresis lanes.

42. A fluorescent electrophoresis apparatus comprising:

a plurality of capillaries having respective ends disposed along a straight line, the capillaries each containing a gel through which a sample labeled with a fluorophore migrates until the sample labeled with the fluorophore is eluted from the end of the respective capillary;

a plurality of migration portions following respective ones of the capillaries and being disposed along a straight line, wherein the sample labeled with the fluorophore eluted from the end of each of the capillaries migrates through a respective one of the migration portions;

a light source for generating light which passes through the migration portions along a straight line extending through all of the migration portions, thereby irradiating the migration portions at respective irradiated portions disposed along the straight line extending through all of the migration portions;

a plurality of optical fibers having respective first ends and respective second ends, the first ends of the optical fibers being disposed facing respective ones of the irradiated portions of the migration portions so as to receive fluorescence generated in the irradiated portions and such that respective optical axes of the optical fibers extending from the first ends of the optical fibers are substantially perpendicular to the straight line extending through all of the migration portions, wherein the optical fibers transmit the fluorescence received by the first ends of the optical fibers to the second ends of the optical fibers, thereby causing the fluorescence to be emitted from the second ends of the optical fibers; and

optical detecting means optically coupled to the second ends of the optical fibers for detecting the fluorescence emitted from the second ends of the optical fibers, thereby detecting the sample labeled with the fluorophore eluted from the end of each of the capillaries outside the capillaries.

43. A fluorescent electrophoresis apparatus comprising:

a plurality of optical cells;

a plurality of capillaries having respective ends disposed along a straight line inside the optical cells, the capillaries each containing a gel through which a sample labeled with a fluorophore migrates until the sample labeled with the fluorophore is eluted from the end of the respective capillary;

a plurality of migration portions formed in the optical cells, the migration portions following respective ones of the capillaries and being disposed along a straight line in the optical cells, wherein the sample labeled with the fluorophore eluted from the end of each of the capillaries migrates through a respective one of the migration portions;

a single light source for generating light which passes through the migration portions in the optical cells along a straight line extending through all of the migration portions, thereby irradiating the migration portions at respective irradiated portions disposed along the straight line extending through all of the migration portions;

a plurality of optical fibers having respective first ends and respective second ends, the first ends of the optical fibers being disposed facing respective ones of the irradiated portions of the migration portions so as to receive fluorescence generated in the irradiated portions and such that respective optical axes of the optical fibers extending from the first ends of the optical fibers intersect the straight line extending through all of the migration portions at respective points where the sample labeled with the fluorophore migrating through the respective one of the migration portions crosses the straight line extending through all of the migration portions, wherein the optical fibers transmit the fluorescence received by the first ends of the optical fibers to the second ends of the optical fibers, thereby causing the fluorescence to be emitted from the second ends of the optical fibers; and

a plurality of optical detecting means optically coupled to the second ends of the optical fibers for detecting the fluorescence emitted from the second ends of the optical fibers, thereby detecting the sample labeled with the fluorophore eluted from the end of each of the capillaries outside the capillaries.

44. A fluorescent electrophoresis apparatus comprising:

a plurality of electrophoresis lanes having respective ends disposed along a straight line, the electrophoresis lanes each containing a separation medium through which a sample labeled with a fluorophore migrates until the sample labeled with the fluorophore is eluted from the end of the respective electrophoresis lane;

means for forming a plurality of sheath flows along respective lines extended from the ends of the electrophoresis lanes, the sheath flows being disposed along a straight line, wherein the sample labeled with the fluorophore eluted from the end of each of the electrophoresis lanes moves in a respective one of the sheath flows;

a light source for generating light which passes through the sheath flows along a straight line extending through all of the sheath flows, thereby simultaneously irradiating the sheath flows at respective irradiated portions disposed along the straight line extending through all of the sheath flows;

a plurality of optical fibers having respective first ends and respective second ends, the first ends of the optical fibers being disposed facing respective ones of the irradiated portions of the sheath flows so as to receive fluorescence generated in the irradiated portions, wherein the optical fibers transmit the fluorescence received by the first ends of the optical fibers to the second ends of the optical fibers, thereby causing the fluorescence to be emitted from the second ends of the optical fibers; and

optical detecting means optically coupled to the second ends of the optical fibers for detecting the fluorescence emitted from the second ends of the optical fibers, thereby detecting the sample labeled with the fluorophore eluted from the end of each of the electrophoresis lanes outside the electrophoresis lanes.

45. A fluorescent electrophoresis apparatus comprising:

a plurality of capillaries having respective ends disposed along a straight line, the capillaries each containing a gel through which a sample labeled with a fluorophore migrates until the sample labeled with the fluorophore is eluted from the end of the respective capillary;

means for forming a plurality of sheath flows along respective lines extended from the ends of the capillaries, the sheath flows being disposed along a straight line, wherein the sample labeled with the fluorophore eluted from the end of each of the capillaries moves in a respective one of the sheath flows;

a light source for generating light which passes through the sheath flows along a straight line extending through all of the sheath flows, thereby irradiating the sheath flows at respective irradiated portions disposed along the straight line extending through all of the sheath flows;

a plurality of optical fibers having respective first ends and respective second ends, the first ends of the optical fibers being disposed facing respective ones of the irradiated portions of the sheath flows so as to receive fluorescence generated in the irradiated portions and such that respective optical axes of the optical fibers extending from the first ends of the optical fibers are substantially perpendicular to the straight line extending through all of the sheath flows, wherein the optical fibers transmit the fluorescence received by the first ends of the optical fibers to the second ends of the optical fibers, thereby causing the fluorescence to be emitted from the second ends of the optical fibers; and

optical detecting means optically coupled to the second ends of the optical fibers for detecting the fluorescence emitted from the second ends of the optical fibers, thereby detecting the sample labeled with the fluorophore eluted from the end of each of the capillaries outside the capillaries.

46. A fluorescent electrophoresis apparatus comprising:

a plurality of optical cells;

a plurality of capillaries having respective ends disposed along a straight line inside the optical cells, the capillaries each containing a gel through which a sample labeled with a fluorophore migrates until the sample labeled with the fluorophore is eluted from the end of the respective capillary;

means for forming a plurality of sheath flows in the optical cells along respective lines extended from the ends of the capillaries, the sheath flows being disposed along a straight line in the optical cells, wherein the sample labeled with the fluorophore eluted from the end of each of the capillaries moves in a respective one of the sheath flows;

a single light source for generating light which passes through the sheath flows in the optical cells along a straight line extending through all of the sheath flows, thereby irradiating the sheath flows at respective irradiated portions disposed along the straight line extending through all of the sheath flows;

a plurality of optical fibers having respective first ends and respective second ends, the first ends of the optical fibers being disposed facing respective ones of the irradiated portions of the sheath flows so as to receive fluorescence generated in the irradiated portions and such that respective optical axes of the optical fibers extending from the first ends of the optical fibers intersect the straight line extending through all of the sheath flows at respective points where the sample labeled with the fluorophore moving in the respective one of the sheath flows crosses the straight line extending through all of the sheath flows, wherein the optical fibers transmit the fluorescence received by the first ends of the optical fibers to the second ends of the optical fibers, thereby causing the fluorescence to be emitted from the second ends of the optical fibers; and

a plurality of optical detecting means optically coupled to the second ends of the optical fibers for detecting the fluorescence emitted from the second ends of the optical fibers, thereby detecting the sample labeled with the fluorophore eluted from the end of each of the capillaries outside the capillaries.