Histological specimen treatment apparatus and method

• US 20090298172A1
• Filed: 05/28/2008
• Published: 12/03/2009
• Est. Priority Date: 05/28/2008
• Status: Abandoned Application

• Current Assignee: Steven Paul Wheeler
• Original Assignee: Steven Paul Wheeler
• Inventors: Wheeler, Steven Paul
• Application Number:
  US12/128,523
• Publication Number:
  US 20090298172A1
• Time in Patent Office: Days
• US Class Current: 435/374
• CPC Class Codes: G01N 1/31 Apparatus therefor
• CPC Class Codes: G01N 1/36 Embedding or analogous mounting of samples

Abstract

Method and apparatus for processing tissue specimens against decomposition, putrefaction and autolysis which use a simple three step procedure in a single vessel or container. First, the specimens are saturated with a solvent mixture of a ketone and a hydrocarbon, e.g., an Acetone/Hexane or an Acetone/Xylene mixture, to dissolve lipids and other cellular solutes. The container is then flooded with melted Paraffin. In a last step, the solvent mixture is vaporized and evacuated from the container, allowing the melted Paraffin to replace the vaporized solvent and impregnate the specimens. Raw, i.e., non-processed and non-burred specimens up to 5 mm thick can be processed in about 60 minutes. A solvent regenerator distills the evacuated solvent; and converts vent waste gases into carbon dioxide and water through a thermocatalytic oxidizer.

Claims

1. A tissue processing method which comprises:

providing a pressure and temperature controllable reaction container;

placing at least one fresh tissue specimen in said container;

treating said specimen in said container to remove solutes and replace said solutes with Paraffin, said step of treating consisting essentially of;

introducing a temperature and pressure conditioned dissolving compound into said container;

allowing time for said compound to penetrate said specimen and to dilute solutes therein;

flooding said container with liquid Paraffin;

vaporizing said compound and allowing a period for said Paraffin to impregnate said specimens;

evacuating said compound and diluted solutes from said container; and

reducing the temperature in said container below the melting point of Paraffin.

2. The method of claim 1, wherein said dissolving compound comprises a mixture taken from the group consisting of azeotropic and zeotropic dehydrant mixtures.

3. The method of claim 1, wherein said dissolving compound comprises a solvent taken from a group consisting essentially of ketones, esters, alcohols, aldehydes, ethers, aromatics, low molecular weight hydrocarbons, and a mixture of low molecular weight hydrocarbons.

4. The method of claim 1, wherein said dissolving compound comprises a mixture taken from a group consisting essentially of:

a mixture of about 60% per weight Acetone and about 40% per weight of at least one Hexane;

a mixture of about 46% per weight Acetone, about 25% per weight of at least one Hexane, about 7% per weight DMSO, about 20% per weight 2-Propanol and about 2% per weight Paraffin;

a mixture of about 62% per weight Acetone, about 16% per weight of at least one Hexane, about 4% per weight DMSO, about 12% per weight 2-Propanol and about 6% per weight Paraffin;

a mixture of about 17% per weight Acetone, about 39% per weight of at least one Hexane, about 5% per weight DMSO, about 12% per weight 2-Propanol and about 30% per weight Paraffin;

a mixture of about 46% per weight Acetone, about 25% per weight of at least one Hexane, about 7% per weight DMSO and about 20% per weight 2-Propanol;

a mixture of about 53% per weight Acetone, about 26% per weight at least one Xylene, about 20% per weight Ethanol, and about 1% per weight DMSO; and

,

a mixture of about 62% per weight Acetone, about 19% per weight at least one Xylene, and about 19% per weight Ethanol.

5. The method of claim 1, wherein said dissolving compound consists of a mixture taken from a group consisting essentially of:

a mixture of about 60% per weight Acetone and about 40% per weight of at least one Hexane;

a mixture of about 46% per weight Acetone, about 25% per weight of at least one Hexane, about 7% per weight DMSO, about 20% per weight 2-Propanol and about 2% per weight Paraffin;

a mixture of about 62% per weight Acetone, about 16% per weight of at least one Hexane, about 4% per weight DMSO, about 12% per weight 2-Propanol and about 6% per weight Paraffin;

a mixture of about 17% per weight Acetone, about 39% per weight of at least one Hexane, about 5% per weight DMSO, about 12% per weight 2-Propanol and about 30% per weight Paraffin;

a mixture of about 46% per weight Acetone, about 25% per weight of at least one Hexane, about 7% per weight DMSO and about 20% per weight 2-Propanol;

a mixture of about 53% per weight Acetone, about 26% per weight at least one Xylene, about 20% per weight Ethanol, and about 1% per weight DMSO; and

,

a mixture of about 62% per weight Acetone, about 19% per weight at least one Xylene, and about 19% per weight Ethanol.

6. The method of claim 1, which further comprises;

regenerating an amount of said dissolving compound evacuated from said container in a regeneration canister.

7. The method of claim 6, wherein said regenerating comprises:

distilling said evacuated compound; and

converting vent waste gases into carbon dioxide and water through a thermocatalytic oxidizer.

8. The method of claim 6, which further comprises conditioning regenerated dissolving compound for introduction into said reaction container.

9. The method of claim 8, wherein said conditioning comprises:

heating said dissolving compound in a solvent vessel to a temperature of approximately 60°

C.; and

establishing a pressure of approximately 0.8 bars (12 psig) in said vessel.

10. The method of claim 1, wherein said time is limited to no more than approximately 30 minutes, and said period is limited to no more than approximately 30 minutes.

11. The method of claim 10, wherein said specimens are taken from a group consisting essentially of non-processed and non-burred tissue sections up to 5 mm thick.

12. The method of claim 1, wherein:

said time is limited to no more than approximately 30 minutes, and said period is limited to no more than approximately 50 minutes; and

said specimens comprise non-processed and non-burred tissue sections up to 10 mm thick.

13. The method of claim 1, which further comprises admitting a limited amount of Paraffin into said container before introducing said dissolving compound.

14. The method of claim 1, wherein said dissolving compound has a refractive index between about 1.350 and 1.499 at 20°

C.

15. The method of claim 1, wherein said dissolving compound comprises a lubricant for process pumps and valves.

16. The method of claim 1, wherein said dissolving compound comprises a mixture of approximately 17% to 65% per weight Acetone and 16% to 40% per weight at least one Hexane.

17. The method of claim 1, wherein said dissolving compound is vaporizable at a pressure of no more than 10 torr and a temperature of no more than about 80°

C.

18. The method of claim 17, wherein said dissolving compound has a refractive index between about 1.350 and 1.499 at 20°

C.

19. A histoprocess which comprises:

dissolving and removing cellular solutes in a tissue specimen using a single dissolving compound; and

replacing said solutes with Paraffin;

said process being performed in a continuous sequence of steps within a single vessel.

20. The histoprocess of claim 19, wherein said dissolving compound comprises a mixture taken from the group consisting of azeotropic and zeotropic dehydrant mixtures.

21. The histoprocess of claim 19, wherein said dissolving compound comprises a solvent taken from a group consisting essentially of ketones, esters, alcohols, aldehydes, ethers, aromatics, low molecular weight hydrocarbons, and a mixture of low molecular weight hydrocarbons.

22. The histoprocess of claim 19, wherein said dissolving compound comprises Acetone and at least one Hexane.

23. The histoprocess of claim 19, wherein said steps comprise:

saturating said specimen with said dissolving compound;

flooding said vessel with liquid Paraffin; and

vaporating said dissolving compound and said solutes to allow said Paraffin to infiltrate spaces vacated by said compound and solutes.

24. The histoprocess of claim 19, which further comprises:

evacuating said dissolving compound and solutes to a regeneration canister;

recuperating said dissolving compound by distillation; and

disposing of vent gases by thermocatalytic oxidation.

25. The histoprocess of claim 19, which further comprises mixing said compound in said vessel prior to said dissolving and removing.

26. The histoprocess of claim 19, wherein said dissolving compound comprises a lubricant for process pumps and valves.

27. The histoprocess of claim 19, wherein said dissolving compound has a refractive index between about 1.350 and 1.499 at 20°

C.

28. The histoprocess of claim 19, wherein said dissolving compound comprises a mixture of approximately 17% to 65% per weight Acetone and 16% to 40% per weight at least one Hexane.

29. The histoprocess of claim 19, wherein said dissolving compound is vaporizable at a pressure of no more than 10 torr and a temperature of no more than about 80°

C.

30. The histoprocess of claim 19, wherein:

said time is limited to no more than approximately 30 minutes, and said period is limited to no more than approximately 50 minutes; and

said specimens comprise non-processed and non-burred tissue sections up to 10 mm thick.

31. The histoprocess of claim 19, which further comprises admitting a limited amount of Paraffin into said container before introducing said dissolving compound.

32. An apparatus for processing tissue specimens against decomposition, putrefaction and autolysis which comprises:

a hermetically sealable reaction container;

means for controlling the temperature and pressure inside said container;

a source of dissolving compound connectable to said container;

a source of melted Paraffin connectable to said container; and

means for evacuating gases from said container;

wherein said dissolving compound comprises a mixture of a ketone and at least one low molecular weight hydrocarbon.

33. The apparatus of claim 32, wherein said dissolving compound comprises a mixture taken from the group consisting of azeotropic and zeotropic dehydrant mixtures.

34. The apparatus of claim 32, wherein said dissolving compound comprises a mixture taken from a group consisting essentially of:

a mixture of about 60% per weight Acetone and about 40% per weight at least one Hexane;

a mixture of about 46% per weight Acetone, about 25% per weight at least one Hexane, about 7% per weight DMSO, about 20% per weight 2-Propanol and about 2% per weight Paraffin;

a mixture of about 62% per weight Acetone, about 16% per weight at least one Hexane, about 4% per weight DMSO, about 12% per weight 2-Propanol and about 6% per weight Paraffin;

a mixture of about 17% per weight Acetone, about 39% per weight at least one Hexane, about 5% per weight DMSO, about 12% per weight 2-Propanol and about 30% per weight Paraffin;

a mixture of about 46% per weight Acetone, about 25% per weight at least one Hexane, about 7% per weight DMSO and about 20% per weight 2-Propano;

a mixture of about 53% per weight Acetone, about 26% per weight at least one Xylene, about 20% per weight Ethanol, and about 1% per weight DMSO; and

,

a mixture of about 62% per weight Acetone, about 19% per weight at least one Xylene, and about 19% per weight Ethanol.

35. The apparatus of claim 32, wherein said dissolving compound consists of a mixture taken from a group consisting essentially of:

a mixture of about 60% per weight Acetone and about 40% per weight at least one Hexane;

a mixture of about 46% per weight Acetone, about 25% per weight at least one Hexane, about 7% per weight DMSO, about 20% per weight 2-Propanol and about 2% per weight Paraffin;

a mixture of about 62% per weight Acetone, about 16% per weight at least one Hexane, about 4% per weight DMSO, about 12% per weight 2-Propanol and about 6% per weight Paraffin;

a mixture of about 17% per weight Acetone, about 39% per weight at least one Hexane, about 5% per weight DMSO, about 12% per weight 2-Propanol and about 30% per weight Paraffin;

a mixture of about 46% per weight Acetone, about 25% per weight at least one Hexane, about 7% per weight DMSO and about 20% per weight 2-Propano;

a mixture of about 53% per weight Acetone, about 26% per weight at least one Xylene, about 20% per weight Ethanol, and about 1% per weight DMSO; and

,

a mixture of about 62% per weight Acetone, about 19% per weight at least one Xylene, and about 19% per weight Ethanol.

36. The apparatus of claim 32, wherein Paraffin present in the solvent provides lubricating properties for process pumps and valves.

37. The apparatus of claim 32, wherein said dissolving compound comprises a solvent taken from the group consisting essentially of ketones, esters, alcohols, aldehydes, ethers, aromatics, low molecular weight hydrocarbons, and a mixture of low molecular weight hydrocarbons.

38. The apparatus of claim 32, which further comprises means for regenerating vaporized dissolving compound evacuated from said container.

39. The apparatus of claim 38, wherein said means for regenerating comprise:

a still for recuperating said evacuated dissolving compound; and

a thermocatalytic oxidizer for converting vent waste gases into carbon dioxide and water.

40. The apparatus of claim 32, wherein said dissolving compound has a refractive index between about 1.350 and 1.499 at 20°

C.

41. The apparatus of claim 32, wherein said dissolving compound comprises a mixture of approximately 17% to 65% per weight Acetone and 16% to 40% per weight at least one Hexane.

42. The apparatus of claim 32, wherein:

said time is limited to no more than approximately 30 minutes, and said period is limited to no more than approximately 50 minutes; and

said specimens comprise non-processed and non-burred tissue sections up to 10 mm thick.

43. The combination of a tissue specimen with a preparation for dissolving lipids and other cellular solutes in said specimen, said preparation being taken from a group consisting essentially of:

a mixture of about 60% per weight Acetone and about 40% per weight at least one Hexane;

a mixture of about 46% per weight Acetone, about 25% per weight at least one Hexane, about 7% per weight DMSO, about 20% per weight 2-Propanol and about 2% per weight Paraffin;

a mixture of about 62% per weight Acetone, about 16% per weight at least one Hexane, about 4% per weight DMSO, about 12% per weight 2-Propanol and about 6% per weight Paraffin;

a mixture of about 17% per weight Acetone, about 39% per weight at least one Hexane, about 5% per weight DMSO, about 12% per weight 2-Propanol and about 30% per weight Paraffin;

a mixture of about 46% per weight Acetone, about 25% per weight at least one Hexane, about 7% per weight DMSO and about 20% per weight 2-Propanol;

a mixture of about 53% per weight Acetone, about 26% per weight at least one Xylene, about 20% per weight Ethanol, and about 1% per weight DMSO; and

,

a mixture of about 62% per weight Acetone, about 19% per weight at least one Xylene, and about 19% per weight Ethanol.

44. A process for removing cellular solutes from a tissue specimen which comprises treating said specimen with a dissolving compound having a viscosity of no more than about 1.30 centipoises at 60°

C. and a boiling point of more than about 80°

C. at a pressure of no more than 10 torr.

45. The process of claim 44, wherein said compound is zeotropic.

46. The process of claim 44, wherein said dissolving compound has a viscosity of no more than about 0.75 centipoises at 60°

C. and a boiling point of less than about 80°

C. at a pressure of no more than 10 torr.

47. The process of claim 44, wherein said compound is regenerated after combination with said solutes.

48. The process of claim 44, wherein said dissolving compound is azeotropic.

49. The process of claim 44, wherein said dissolving compound comprises a solvent taken from a group consisting essentially of ketones, esters, alcohols, aldehydes, ethers, aromatics, low molecular weight hydrocarbons, and a mixture of low molecular weight hydrocarbons.

50. The process of claim 44, wherein said dissolving compound comprises a lubricant for process pumps and valves.