Detection of reactions and metabolic changes with fluorscent materials

• US 20030012692A1
• Filed: 05/22/2002
• Published: 01/16/2003
• Est. Priority Date: 05/25/2001
• Status: Abandoned Application

• Current Assignee: Corning Incorporated
• Original Assignee: Corning Incorporated
• Inventors: Marque, Pascal, Pecheul, Marylene D. M., Lemee, Valerie J.C., Root, David M.
• Application Number:
  US10/154,465
• Publication Number:
  US 20030012692A1
• Time in Patent Office: Days
• US Class Current: 422/57
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• CPC Class Codes: B01L 3/50851 specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/78 producing a change of colour
• CPC Class Codes: G01N 2500/00 Screening for compounds of potential therapeutic value
• CPC Class Codes: G01N 33/5005 involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/502 for testing non-proliferative effects
• CPC Class Codes: G01N 33/5029 on cell motility

Abstract

A system, method and device for the detection of reactions between analytes, (e.g., DNA, biomolecules, or cells) and a second compound are disclosed. The present invention includes a coating of a fluorescent material having a fluorescence that changes with temperature. The fluorescent material is associated with a substrate, and can be used for any type of surface reaction that requires determination of temperature conditions at an interface between the surface and the reaction analyte subject to assay. The substrate may be, for example, a microarray chip or microplate, preferably suitable for use in high-throughput screening of biomolecules or cells. Substrates containing the fluorescent material also can be used to compensate for temperature variations in refractive index in optical sensors.

Claims

1. A system for performing high-throughput screening or assays of biological or chemical analytes, the system comprising:

a sample holder with an array of locations each having a bottom surface;

a fluorescent material, the fluorescence of which changes with temperature, in thermal communication with the bottom surface;

a first element for interacting said biological or chemical analytes with a compound in at least one of the array locations; and

a second element for measuring a change in fluorescence of the fluorescent material as the temperature in at least one of the locations changes.

2. The system according to claim 1, wherein the biological analytes include:

biological molecules, cells, cell components, nucleic acids, and proteins.

3. The system according to claim 1, wherein a layer of the fluorescent material is in thermal communication with the biological analytes.

4. The system according to claim 2, wherein a sensing area on the bottom surface is proximate to said biological analytes.

5. The system according to claim 1, wherein the fluorescent material is operative to produce a change in fluorescence sufficient to detect a chemical reaction, a biomolecular reaction, or a metabolic response of a cell.

6. The system according to claim 1, wherein the sample holder is a multi-well plate.

7. The system according to claim 1, wherein the fluorescent material is in the form of a film forming a layer adjacent the bottom surface of a microplate.

8. The system according to claim 6, wherein the layer is on the bottom surface of the multi-well plate.

9. The system according to claim 1, wherein fluorescent material is embedded in the bottom surface of the microplate.

10. The system according to claim 1, wherein the fluorescent material includes a rare-earth chelate.

11. The system according to claim 9, wherein the rare earth chelate is EuTTA.

12. The system according to claim 9, wherein the fluorescent material is selected from the group consisting of EuTTA, Rhodamine B, Erythrosin B, terthiophene, and combinations thereof.

13. The system according to claim 9, wherein the fluorescent material includes EuFOD, EuTFC, TbFOD, EuBA, EuTHD, EuHFC, EuDBM, EuTA, EuTFA, EuDCM, TTED, TbTTA, TbTFA, TbBA, TbTHD, TbAA, and combinations thereof.

14. The system according to claim 3, wherein the layer≦

about 50 microns.

16. The bisosensor according to claim 15, wherein said fluorescent material is in thermal communication with a surface of the substrate.

17. The biosensor according to claim 15, wherein the fluorescent material includes a rare-earth chelate.

18. The biosensor according to claim 17, wherein the rare earth chelate is EuTTA.

19. The biosensor according to claim 17, wherein the fluorescent material is selected from the group consisting of EuTTA, Rhodamine B, Erythrosin B, terthiophene, and combinations thereof.

20. The biosenor according to claim 15, wherein the fluorescent material includes EuFOD, EuTFC, TbFOD, EuBA, EuTHD, EuHFC, EuDBM, EuTA, EuTFA, EuDCM, TTED, TbTTA, TbTFA, TbBA, TbTHD, TbAA, and combinations thereof.

21. The biosensor according to claim 15, wherein the fluorescent material is in a layer≦

about 50 microns.

22. The biosensor according to claim 15, wherein a portion of the capillary adjacent to the biological or chemical analytes is capable of being monitored for changes in temperature.

23. The biosensor according to claim 22, wherein the biological analytes include biomolecules, cells, or cell fragments.

24. The biosensor according to claim 15, wherein changes in temperature of the portion of the capillary adjacent to the biological analytes can be monitored to detect metabolic changes when the biological analytes are cells.

26. The microarray according to claim 25, wherein the fluorescent material includes a rare-earth chelate.

27. The microarray according to claim 26, wherein the rare earth chelate is EuTTA.

28. The microarray according to claim 26, wherein the fluorescent material is selected from the group consisting of EuTTA, Rhodamine B, Erythrosin B, terthiophene, and combinations thereof.

29. The microarray according to claim 25, wherein the fluorescent material includes EuFOD, EuTFC, TbFOD, EuBA, EuTHD, EuHFC, EuDBM, EuTA, EuTFA, EuDCM, TTED, TbTTA, TbTFA, TbBA, TbTHD, TbAA, and combinations thereof.

30. The microarray according to claim 25, wherein the layer is≦

about 50 microns.

31. The microarray according to claim 25, wherein the biological analytes include DNA, RNA, and biomolecules.

15. A biosensor comprising:

a substrate of a porous structure with a fluorescent material, the fluorescence of which changes with changing temperature;

a sample of biological or chemical analytes in close proximity with said substrate;

said substrate including at least one capillary through which the biological or chemical analytes migrate.

25. A microarray comprising biological or chemical analytes disposed on a substrate having a layer of a fluorescent material in thermal communication with a surface of the substrate, wherein the fluorescence of the fluorescent material changes with changing temperature.

32. A device for performing an assay, the device comprising:

a substrate having analytes;

a detecting element to detect a light signal proximate to the sensing area;

measuring elements to measure respectively (a) a temperature proximate the sensing area and (b) a refractive index of the sensing area; and

an adjusting element to adjust the light signal in accordance with a change in refractive index of the sending area due to a change in temperature.

33. The device according to claim 32, wherein the device further comprises a fluorescent material, the fluorescence of which changes with temperature, in thermal communication with the substrate or analytes.

34. The device according to claim 32, wherein said device detects surface reaction that requires determination of temperature conditions at an interface between the surface and the analytes.

35. The device according to claim 33, wherein the fluorescent material includes a rare-earth chelate.

36. The device according to claim 35, wherein the fluorescent material is selected from the group consisting of EuTTA, Rhodamine B, Erythrosin B, terthiophene, and combinations thereof.

37. The device according to claim 35, wherein the rare earth chelate is EuTTA.

38. The device according to claim 33, wherein the fluorescent material includes EuFOD, EuTFC, TbFOD, EuBA, EuTHD, EuHFC, EuDBM, EuTA, EuTFA, EuDCM, TTED, TbTTA, TbTFA, TbBA, TbTHD, TbAA, and combinations thereof.

39. The device according to claim 32, wherein the analytes include biomolecules, cells, or cell fragments.

40. A method for analyzing substances proximate to a sensing area of a surface, the method comprising:

detecting a light signal generated proximate to the sensing area;

measuring the temperature proximate to the sensing area;

measuring the refractive index of the sensing area; and

determining the change in refractive index of the sensing area due to the change in temperature and adjusting the light signal in accordance with the change in refractive index.

41. The method of claim 40, wherein the sensing area includes a substrate and a fluid in contact with the substrate.

42. The method of claim 41, wherein the substrate is selected from the group consisting of a microplate, a micro array and a micro fluidic device.

43. The method of claim 41, wherein the fluid contains a cell or a biomolecule.

44. The method of claim 43, wherein a fluorescent material having a temperature-dependent fluorescence is proximate the sensing area.

45. The method according to claim 44, wherein the fluorescent material includes a rare-earth chelate.

46. The method of claim 45, wherein the fluorescent material is selected from the group consisting of EuTTA, Rhodamine B, Erythrosin B, terthiophene and combinations thereof.

47. The method according to claim 45, wherein the rare earth chelate is EuTTA.

48. The method according to claim 44, wherein the fluorescent material includes EuFOD, EuTFC, TbFOD, EuBA, EuTHD, EuHFC, EuDBM, EuTA, EuTFA, EuDCM, TTED, TbTTA, TbTFA, TbBA, TbTHD, TbAA, and combinations thereof.

49. The method of claim 44, wherein the fluorescent material is embedded in the substrate

50. The method of claim 44, wherein the fluorescent material is in the form of a layer on a surface of the substrate.

51. The method according to claim 50, wherein the fluorescent material is in a layer≦

about 50 microns.

52. The method of claim 41, wherein the substrate includes at least one capillary, and the fluorescent material is adjacent the capillary.

53. A method of detecting a chemical reaction, a biomolecular reaction or a metabolic change in a cell comprising:

providing a substrate including a fluorescent material, the fluorescence of which changes as the temperature of the fluorescent material changes;

interacting a chemical, a biomolecule, or a cell in contact with a second compound in or on the substrate;

monitoring the fluorescence of the fluorescent material; and

correlating the fluorescence of the fluorescent material with a reaction between the biomolecule and the compound.

54. The method of claim 53, further comprising the step of correlating involves comparing the light intensity of the fluorescent material with a change in temperature.

55. The method of claim 53, wherein the substrate is a microplate, microfluidic device or a microarray chip incorporating a fluorescent film.

57. The method of claim 56, wherein the array of locations is provided on a microplate.

58. The method of claim 56, wherein the array of locations includes a microarray chip.

60. The optical sensing system according to claim 59, wherein the sensing area includes a waveguide in association with the substrate.

61. The optical sensing system according to claim 60, wherein the waveguide includes a planar waveguide, a waveguide film, an optical fiber or a grating structure.

62. The optical sensing system according to claim 59, wherein the system includes a photon sieve or an interferometer.

63. The optical sensing system according to claim 59, wherein the processor is operative to receive an optical signal and correlate changes in temperature with changes in the refractive index of the substrate and/or the fluid.

64. The optical sensing system according to claim 59, wherein the processor is operative to adjust the optical signal in accordance with the correlated change in refractive index of the fluid and/or the substrate.

56. A method of screening biochemical assays comprising:

providing biochemical molecules in an array of locations;

interacting a compound in reactive contact with the biochemical molecules in at least one of the locations;

detecting the temperature change in the at least of one of the locations by detecting the change in fluorescence of the locations;

wherein at least one of the array of locations contains a fluorescent material, the fluorescence of which changes with temperature.

59. An optical sensing system comprising:

a substrate in contact with a fluid containing a biomolecule or a cell;

a sensing area proximate to the substrate;

a light signal generated proximate to the sensing area;

a light detector; and

a fluorescent material in association with the substrate, the fluorescence of which changes with changing temperature;

a processor for determining temperature changes corresponding to the changing in fluorescence and changes in refractive index of the sensing area due to the change in temperature and adjusting the light signal in accordance with the change in refractive index.