Claims
- 1. An automated method of removing embedding media from a biological sample on a microscope slide, the method comprising the steps of:
heating the biological sample and the embedding media above the, melting point of the embedding media; and rinsing with an immiscible fluid the heated embedding media from the biological sample.
- 2. The method of claim 1, wherein the step of rinsing includes rinsing the melted embedding media from the biological sample.
- 3. The method of claim 4, wherein the step of heating a bottom side of the slide includes heating the biological sample to temperatures ranging from ambient to 130° C.
- 4. The method of claim 1, wherein the embedding media is paraffin.
- 5. The method of claim 1, wherein the fluid is a gas.
- 6. The method of claim 1, wherein the fluid is a liquid.
- 7. The method of claim 6, wherein the liquid is not an organic solvent.
- 8. The method of claim 6, wherein the liquid is selected from the group consisting of de-ionized water, citrate buffer (pH 6.0-8.0), Tris-HCl buffer (pH 6-10), phosphate buffer (pH 6.0-8.0), SSC buffer, APK Wash™, acidic buffers or solutions (pH 1-6.9), and basic buffers or solutions (pH 7.1-14).
- 9. The method of claim 1, wherein the fluid includes ionic or non-ionic surfactants.
- 10. The method of claim 9, wherein the ionic or non-ionic surfactants are selected from the group consisting of Triton X-100, Tween, Brij, sodium dodecylsulfate and saponin.
- 11. The method of claim 1, wherein the fluid includes a detergent.
- 12. An automated method of removing paraffin from a paraffin-embedded biological sample on a microscope slide, the method comprising the steps of:
heating the paraffin-embedded biological sample; and applying a paraffin-immicible liquid on the biological sample.
- 13. The method of claim 12, wherein the liquid applied has a density which is greater than the liquified paraffin.
- 14. The method of claim 12, wherein the liquid does not solvate the paraffin.
- 15. The method of claim 12, wherein the liquid is selected from the group consisting of de-ionized water, citrate buffer (pH 6.0-8.0), Tris-HCl buffer (pH 6-10), phosphate buffer (pH 6.0-8.0), SSC buffer, APK Wash™, acidic buffers or solutions (pH 1-6.9), and basic buffers or solutions (pH 7.1-14).
- 16. The method of claim 12, wherein the liquid includes ionic or non-ionic surfactants.
- 17. The method of claim 16, wherein the ionic or non-ionic surfactants are selected from the group consisting of Triton X-100, Tween, Brij, sodium dodecylsulfate and saponin.
- 18. The method of claim 12, wherein the liquid includes a detergent.
- 19. The method of claim 12, wherein the step of applying a liquid is performed during the step of heating the paraffin-embedded biological sample.
- 20. The method of claim 12, wherein the step of heating the paraffin-embedded biological sample includes the following steps:
heating the paraffin-embedded biological sample without liquid on the biological sample; and heating the paraffin-embedded biological sample with liquid on the biological sample, whereby the step of heating the paraffin-embedded biological sample without liquid on the paraffin-embedded biological sample removes moisture between the paraffin-embedded biological sample and the surface of the slide.
- 21. The method of claim 20, wherein the step of heating the paraffin-embedded biological sample without liquid on the biological sample melts at least a portion of the paraffin.
- 22. The method of claim 20, wherein the liquid applied is more dense than the paraffin, and
wherein the step of heating the paraffin-embedded biological sample with liquid on the paraffin-embedded biological sample melts at least a portion of the paraffin and causes the paraffin to float to the top of the liquid.
- 23. The method of claim 12, wherein the heating of the biological sample and the paraffin melts at least a portion of the paraffin, and further comprising the step of applying a fluid to remove the at least a portion of the paraffin.
- 24. The method of claim 23, wherein the step of applying a fluid includes rinsing the melted paraffin from the paraffin-embedded biological sample with the fluid.
- 25. The method of claim 24, wherein the fluid is not an organic solvent.
- 26. An automated method of cell conditioning without the removal or etching of the paraffin within a biological staining procedure, the method comprising the steps of:
applying heat to the biological sample; applying at least one conditioning reagent; and applying fluid to remove the at least one conditioning reagent.
- 27. An automated method according to claim 26 wherein the biological sample is on a top surface of a slide; and
wherein the step of heating includes heating a bottom side of the slide.
- 28. An automated method according to claim 27 wherein the bottom side of the slide is in contact with a thermal platform and wherein the step of heating a bottom side of the slide includes heating the slide by conduction using the thermal platform.
- 29. An automated method according to claim 26 wherein the biological sample is heated to temperatures ranging from ambient to 130° C.
- 30. An automated method according to claim 26 wherein the at least one conditioning reagent is selected from the group consisting of air, de-ionized water, citrate buffer (pH 6.0-8.0), Tris-HCl buffer (pH 6-10), phosphate buffer (pH 6.0-8.0), SSC buffer, APK Wash™, acidic buffers or solutions (pH 1-6.9), and basic buffers or solutions (pH 7.1-14).
- 31. An automated method according to claim 26 wherein the at least one conditioning reagent contains ionic or non-ionic surfactants selected from the group consisting of Triton X-100, Tween, Brij, sodium dodecylsulfate and saponin.
- 32. An automated method of simultaneously removing embedding medium from a embedded biological sample while providing cell conditioning within a biological staining procedure, the method comprising the steps of:
applying exposing and cell conditioning reagents; applying heat to the embedded biological sample; applying fluid to remove the exposing and cell conditioning reagents; and staining the biological sample.
- 33. An automated method according to claim 32 wherein the embedded biological sample is on a top surface of a slide; and
wherein the step of heating includes heating a bottom side of the slide.
- 34. An automated method according to claim 33 wherein the bottom side of the slide is in contact with a thermal platform and wherein the step of heating a bottom side of the slide includes heating the slide by conduction using the thermal platform.
- 35. An automated method according to claim 32 wherein the step of applying heat includes heating the biological sample to temperatures ranging from ambient to 130° C.
- 36. An automated method according to claim 32 wherein the exposing and cell conditioning reagents are selected from the group consisting of air, de-ionized water, citrate buffer (pH 6.0-8.0), Tris-HCl buffer (pH 6-10), phosphate buffer (pH 6.0-8.0), SSC buffer, APK Wash™, acidic buffers or solutions (pH 1-6.9), and basic buffers or solutions (pH7.1-14).
- 37. An automated method according to claim 32 wherein the exposing and cell conditioning reagents contain ionic or non-ionic surfactants selected from the group consisting of Triton X-100, Tween, Brij, sodium dodecylsulfate and saponin.
- 38. An automated method of removing or etching embedding media from a embedded biological sample and subsequently providing cell conditioning within a biological staining procedure, the method comprising the steps of:
applying heat to the embedded biological sample; applying a first fluid to the embedded biological sample to remove the embedding media or etching reagents; applying cell conditioning reagents; applying a second fluid to remove the cell conditioning reagents; and staining of the biological sample.
- 39. An automated method according to claim 38 wherein the biological sample is on a top surface of a slide; and
wherein the step of heating includes heating a bottom side of the slide.
- 40. An automated method according to claim 39 wherein the bottom side of the slide is in contact with a thermal platform and wherein the step of heating a bottom side of the slide includes heating the slide by conduction using the thermal platform.
- 41. An automated method according to claim 38 wherein the step of applying heat includes heating the biological sample to temperatures ranging from ambient to 130° C.
- 42. An automated method according to claim 38 further comprising the step of applying exposing reagents to the biological sample
- 43. An automated method according to claim 42 wherein the exposing reagents are selected from the group consisting of air, de-ionized water, citrate buffer (pH 6.0-8.0), Tris-HCl buffer (pH 6-10), phosphate buffer (pH 6.0-8.0), SSC buffer, APK Wash™, acidic buffers or solutions (pH 1-6.9), and basic buffers or solutions (pH7-1-14).
- 44. An automated method according to claim 42 wherein the exposing reagents contain ionic or non-ionic surfactants selected from the group consisting of Triton X-100, Tween, Brij, sodium dodecylsulfate and saponin.
- 45. A composition comprising a buffer, wherein the composition is selected from the group consisting of:
a) 2×SSC; b) 10 mM phosphate buffer with about 0 to about 0.1% Triton-X100; c) deionized water having from about 0 to about 0.1% Triton-X100; d) about 5 to about 50 mM sodium citrate buffer having about 0 to about 0.1% Triton-X100, and about 0 to about 0.5% sodium dodecyl sulfate (pH adjusted to between about 6 and about 8); and e) about 5 to about 20 mM Tris buffer with about 0 to about 0. 1% Triton-X100.
- 46. A composition comprising a buffer, wherein the composition is selected from the group consisting of:
a) about 5 to about 20 mM Tris-HCl, about 0 to about 40 mM boric acid, about 0 to about 2 mM EDTA, about 0 to about 2 mM EDTA, about 0 to about 20% DMSO, about 0 to about 0.5% Brij 35, and about 0 to about 0.1% Triton X100, (pH adjusted from about 7 to about 9); b) about 5 to about 50 mM Citrate buffer, from about 0 to about 0.5% SDS, about 0 to about 10% ethylene glycol, about 0 to about 1 M urea, about 0 to about 20% formamide, about 0 to about 10% DMSO, about 0 to about 0.5% Brij 35, and about 0 to about 0.1% Triton X100 (pH adjusted from about 6 to about 8); c) about 1 to about 50 mM EDTA, about 0 to about 0.75% SDS, about 0 to about 10% ethylene glycol (pH adjusted from about 7 to about 8); d) about 10 mM sodium citrate, about 1.4 mM MgCl2, and about 0.1% SDS (pH adjusted from about 7 to about 8); e) about 10 mM phosphate, and from about 0 to about 0.1% Triton X100 (pH adjusted from about 6 to about 8); f) 2×SSC; and g) about 10 mM phosphate buffer, about 0 to about 10 mM sodium citrate, about 0 to about 5×SSC, and from about 0 to about 2.5% Chondroiton A (pH adjusted from about 7 to about 9).
REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of priority from International Application No. PCT/US99/20353, filed Sep. 3, 1999, which was published on Mar. 16, 2000 as WO 00/14507. International Application No. PCT/US99/20353, in turn, claimed the benefit of priority from U.S. Provisional Patent Application No. 60/099,018, filed Sep. 3, 1998; U.S. patent application Ser. No. 09/259,240, filed Feb. 26, 1999; and International Application No. PCT/US99/04181, filed Feb. 26, 1999, which was published on Sep. 2, 1999 as WO. 99/44030. All of the patent applications mentioned herein are hereby incorporated by reference.
Continuations (1)
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Number |
Date |
Country |
Parent |
09721096 |
Nov 2000 |
US |
Child |
10320219 |
Dec 2002 |
US |