High quality, continuous throughput, tissue processing

• US 7,547,538 B2
• Filed: 03/27/2003
• Issued: 06/16/2009
• Est. Priority Date: 08/20/1997
• Status: Expired due to Term

• Current Assignee: University of Miami
• Original Assignee: University of Miami
• Inventors: Essenfeld, Ervin, Essenfeld, Harold, Morales, Azorides
• Primary Examiner(s): Redding; David A
• Application Number:
  US10/400,060
• Publication Number:
  US 20040004075A1
• Time in Patent Office: 2,273 Days
• Field of Search: 435/283.1, 435/284.1
• US Class Current: 435/284.1
• CPC Class Codes: B01L 9/50 Clamping means, tongs in general F16B2/06
• CPC Class Codes: G01N 1/06 providing a thin slice, e.g. microtome
• CPC Class Codes: G01N 1/30 Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
• CPC Class Codes: G01N 1/312 for samples mounted on planar substrates
• CPC Class Codes: G01N 1/36 Embedding or analogous mounting of samples
• CPC Class Codes: G01N 1/44 Sample treatment involving radiation, e.g. heat
• CPC Class Codes: G01N 2001/305 Fixative compositions
• CPC Class Codes: G01N 2001/315 Basket-type carriers for tissues

Abstract

A process and apparatus for rapid, continuous flow histological processing of tissues is disclosed. The steps of fixation, dehydration, clearing and impregnation are performed in less than one hour; this allows a pathologist to evaluate samples shortly after receipt, perhaps while the patient is still in the operating room. Rapid and continuous processing is accomplished by decreasing the thickness of tissue sections, use of nonaqueous solutions composed of admixtures of solutions, solution exchange at elevated temperature and with agitation, and impregnation under vacuum pressure. The patient in surgery is thus provided with point-of-care surgical pathology.

Claims

1. An apparatus for continuous processing of tissue specimens comprising:

(a) a microwave processor unit which (i) fixes and dehydrates tissue specimens with a non-aqueous solution in a first vessel, (ii) agitates the non-aqueous solution, and (iii) heats the non-aqueous solution with microwave energy to produce processed tissue specimens; and

a first source for the non-aqueous solution which is fluidly coupled to the first vessel; and

(b) an impregnator unit which (i) impregnates processed tissue specimens with a wax solution in a second vessel and (ii) heats the wax solution under less than atmospheric pressure to produce processed and impregnated tissue specimens; and

a second source for the wax solution which is fluidly coupled to the second vessel.

2. The apparatus of claim 1, wherein processing of a subsequent tissue specimen is initiated before impregnation of an earlier tissue specimen has been completed.

3. The apparatus of claim 1, wherein the non-aqueous solution is comprised of an admixture of a ketone and an alcohol, and the volume ratio between alcohol and ketone is from about 1:

1 to about 3;

1.

4. The apparatus of claim 3, wherein the non-aqueous solution comprises about 25% acetone, about 55% isopropyl alcohol, and about 5% mineral oil.

5. The apparatus of claim 1, wherein a tissue specimen is fixed, dehydrated and impregnated in a total time of less than about two hours.

6. The apparatus of claim 1, wherein the tissue specimen is fixed and dehydrated in a series of non-aqueous solutions.

7. The apparatus of claim 1, wherein the tissue specimen is impregnated in a series of paraffin solutions.

8. The apparatus of claim 1, wherein the tissue specimen is fixed and dehydrated in a series of non-aqueous solutions, and impregnated in a series of paraffin solutions.

9. The apparatus of claim 1, wherein the non-aqueous solution comprises acetone, isopropyl alcohol, and mineral oil.

10. The apparatus of claim 1, wherein the volume ratio between alcohol and ketone is from about 1:

1 to about 6;

1.

11. The apparatus of claim 1, wherein the wax solution comprises mineral oil and melted paraffin.

12. The apparatus of claim 1, wherein the wax solution comprises about 30% mineral oil and about 70% melted paraffin.

13. The apparatus of claim 1, wherein the wax solution is degassed.

14. The apparatus of claim 1, wherein the wax solution comprises about 30% mineral oil and about 70% melted paraffin.

15. An apparatus for hardening and impregnating a tissue specimen comprising:

(a) a microwave unit comprising (i) a non-aqueous solution in a first vessel configured to harden the tissue specimen, (ii) a microwave energy source configured to heat, and (iii) an agitator configured to agitate the non-aqueous solution in the first vessel;

wherein the non-aqueous solution in the first vessel is comprised of a ketone and an alcohol; and

(b) a vacuum unit comprising (i) a wax solution in a second vessel configured to harden the tissue specimen, and (ii) a heater configured to heat under less than atmospheric pressure the wax solution in the second vessel;

wherein the wax solution in the second vessel is comprised of paraffin wax; and

wherein the microwave unit and the vacuum unit are combined in the apparatus such that tissue specimens are sequentially transferred into the first vessel and then into the second vessel.

16. The apparatus of claim 15, wherein hardening of a subsequent, not hardened tissue specimen is initiated in the microwave unit before impregnation of an earlier, hardened tissue specimen in the vacuum unit has been completed.

17. The apparatus of claim 15, wherein the volume ratio between alcohol and ketone is from about 1:

1 to about 3;

1.

18. The apparatus of claim 17, wherein the non-aqueous solution comprises about 19% acetone, about 55% isopropyl alcohol, and about 5% mineral oil.

19. The apparatus of claim 15, wherein the tissue specimen is hardened and impregnated in a total time of less than about two hours.

20. The apparatus of claim 15, wherein the tissue specimen is hardened in a series of non-aqueous solutions.

21. The apparatus of claim 15, wherein the tissue specimen is impregnated in a series of paraffin solutions.

22. The apparatus of claim 15, wherein the tissue specimen is hardened in a series of non-aqueous solutions, and impregnated in a series of paraffin solutions.

23. The apparatus of claim 15, wherein the non-aqueous solution comprises acetone, isopropyl alcohol, and mineral oil.

24. The apparatus of claim 15, wherein the volume ratio between alcohol and ketone is from about 1:

1 to about 6;

1.

25. The apparatus of claim 15, wherein the wax solution comprises mineral oil and melted paraffin.

26. The apparatus of claim 15, wherein the wax solution is degassed.