Claims
- 1. A method for preparing a tissue section comprising:
a) contacting a tissue section with a first set of one or more solutions comprising no more than about 50% water; b) contacting the tissue section with a stain in a solution comprising no more than about 50% water; c) then contacting the tissue section with a second set of one or more solutions comprising no more than 50% water, wherein the tissue section is not contacted with a solution that is more than about 50% water.
- 2. The method of claim 1, wherein the tissue section is not contacted with a solution that is more than about 80% water.
- 3. The method of claim 2, wherein the tissue section is not contacted with a solution that is more than about 90% water.
- 4. The method of claim 3, wherein the tissue section is not contacted with a solution that is about 100% water.
- 5. The method of claim 4, wherein the stain is in a solution with less than about 10% water.
- 6. The method of claim 1, wherein one or more of the solutions is at least about 70% alcohol.
- 7. The method of claim 6, wherein all of the solution are at least about 70% alcohol.
- 8. The method of claim 7, wherein one or more of the solutions is at least about 90% alcohol.
- 9. The method of claim 8, wherein one or more of the solutions is about 100% alcohol.
- 10. The method of claim 6, wherein the stain is in a solution that is at least about 80% alcohol.
- 11. The method of claim 10, wherein the stain is in a solution that is about 100% alcohol.
- 12. The method of claim 6, wherein the alcohol in at least one of the solutions comprising alcohol is selected from the group consisting of ethanol, methanol, and propanol.
- 13. The method of claim 1, wherein solutions comprising water include water that been treated with DEPC.
- 14. The method of claim 1, wherein at least one of the solutions in the second set comprises a non-alcohol organic solvent.
- 15. The method of claim 14, wherein the organic solvent solution is at least about 50% organic solvent.
- 16. The method of claim 15, wherein the organic solvent solution is at least about 75% organic solvent.
- 17. The method of claim 16, wherein the organic solvent solution is at least about 90% organic solvent.
- 18. The method of claim 17, wherein the organic solvent solution is about 100% organic solvent.
- 19. The method of claim 14, wherein the organic solvent is a hydrocarbon selected from the group consisting of xylene or toluene.
- 20. The method of claim 1, wherein the stain is selected from the group consisting of cresyl violet acetate, hematoxylin, and eosin.
- 21. The method of claim 1, wherein the tissue sample is exposed to more than one stain.
- 22. The method of claim 1, further comprising performing laser capture microscopy on the tissue sample.
- 23. The method of claim 22, further comprising extracting nucleic acid molecules from cells of the tissue sample that have been subjected to laser capture microdissection.
- 24. The method of claim 23, wherein the nucleic acid molecules include RNA.
- 25. The method of claim 1, further comprising extracting nucleic acid molecules from the tissue sample.
- 26. The method of claim 25, wherein the nucleic acid molecules include RNA.
- 27. A method for preparing a tissue section comprising:
a) contacting a tissue section with a first set of one or more solutions comprising alcohol; b) contacting the tissue section with a stain in a solution comprising an organic solvent; c) contacting the tissue section with a second set of one or more solutions comprising alcohol, wherein the tissue section is not contacted with a solution that does not comprise alcohol or an organic solvent.
- 28. The method of claim 27, wherein the sets of solutions comprising alcohol are at least about 70% alcohol.
- 29. The method of claim 28, wherein one or more of the solutions in each set of solutions comprising alcohol is at least about 90% alcohol.
- 30. The method of claim 29, wherein one or more of the solutions in each set of solutions comprising alcohol is about 100% alcohol.
- 31. The method of claim 27, wherein the stain is in a solution that is at least about 50% organic solvent.
- 32. The method of claim 27, wherein the organic solvent in the stain-containing solution is an alcohol.
- 33. The method of claim 32, wherein the stain is in a solution that is at least about 50% alcohol.
- 34. The method of claim 33, wherein the stain is in a solution that is at least about 80% alcohol.
- 35. The method of claim 27, wherein the stain is in a solution that is about 100% alcohol.
- 36. The method of claim 27, wherein solutions comprising water include water that been treated with DEPC.
- 37. The method of claim 27, further comprising contacting the tissue section to a solution comprising a non-alcohol organic solvent subsequent to exposing the tissue section to a stain.
- 38. The method of claim 37, wherein the organic solvent solution is at least about 50% organic solvent.
- 39. The method of claim 38, wherein the organic solvent solution is at least about 75% organic solvent.
- 40. The method of claim 39, wherein the organic solvent solution is at least about 90% organic solvent.
- 41. The method of claim 40, wherein the organic solvent solution is about 100% organic solvent.
- 42. The method of claim 27, wherein the organic solvent is a hydrocarbon selected from the group consisting of xylene or toluene.
- 43. The method of claim 27, wherein the alcohol in at least one of the solutions comprising alcohol is selected from the group consisting of ethanol, methanol, and propanol.
- 44. The method of claim 27, wherein the stain is selected from the group consisting of cresyl violet acetate, hematoxylin, and cosin.
- 45. The method of claim 27, wherein the tissue sample is exposed to more than one stain.
- 46. The method of claim 27, further comprising performing laser capture microscopy on the tissue sample.
- 47. The method of claim 46, further comprising extracting nucleic acid molecules from cells of the tissue sample that have been subjected to laser capture microdissection.
- 48. The method of claim 47, wherein the nucleic acid molecules include RNA.
- 49. The method of claim 27, further comprising extracting nucleic acid molecules from the tissue sample.
- 50. The method of claim 49, wherein the nucleic acid molecules include RNA.
- 51. A method for preparing a tissue section comprising:
a) contacting the tissue section with a first set of one or more solutions comprising alcohol; b) contacting the tissue section with a stain in a solution in which RNase activity is substantially reduced; and, c) contacting the tissue section with a second set of one or more solutions comprising alcohol.
- 52. A method for preparing a tissue section for RNA extraction comprising:
a) contacting the tissue section in a first solution comprising at least about 70% alcohol; b) exposing the tissue section to second solution comprising a stain and at least about 90% alcohol; c) contacting the tissue section in a third solution comprising at least about 70% alcohol; d) contacting the tissue section in a fourth solution comprising at least about 95% alcohol; e) contacting the tissue section in a fifth solution comprising about 100% alcohol f) contacting the tissue section in sixth solution comprising xylene; g) extracting RNA from the tissue section.
- 53. A kit for preparing a tissue section comprising, in suitable container means:
a) a series of dehydrating solutions comprising an alcohol, wherein any water in the solutions has been treated with DEPC; b) a stain solution comprising a stain dissolved in an organic-solvent containing solution.
- 54. The kit of claim 53, wherein the series of dehydrating solutions comprises at least a first solution comprising at least about 70% alcohol, a second solution comprising at least about 95% alcohol, and a third solution comprising about 100% alcohol.
- 55. The kit of claim 53, further comprising a xylene solution.
Government Interests
[0001] The government may own rights in the present invention pursuant to grant number R43-CA88699 from National Institutes of Health/National Cancer Institute.