High quality, continuous throughput, tissue processing

• US 8,221,996 B2
• Filed: 03/12/2008
• Issued: 07/17/2012
• Est. Priority Date: 08/20/1997
• Status: Expired due to Term

• Current Assignee: University of Miami
• Original Assignee: University of Miami
• Inventors: Morales, Azorides, Essenfeld, Harold, Essenfeld, Ervin
• Primary Examiner(s): Weber, Jon P
• Assistant Examiner(s): UNDERDAHL, THANE E
• Application Number:
  US12/046,465
• Publication Number:
  US 20080153127A1
• Time in Patent Office: 1,588 Days
• Field of Search: 435/40.5
• US Class Current: 435/40.5
• 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 non-aqueous 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. A non-aqueous solution, wherein the non-aqueous solution comprises (i) a ketone, (ii) an alcohol, and (iii) mineral oil;

wherein the volume ratio of alcohol to ketone is from about 1;

1 to about 6;

1.

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

1 to about 3;

1.

3. The solution of claim 2 which further comprises (iv) polyethylene glycol (PEG) and (v) surfactant.

4. The solution of claim 3, wherein said PEG is between about 100 and about 500 average molecular weight.

5. The solution of claim 3, wherein said surfactant is DMSO.

6. The solution of claim 2 which further comprises (iv) polyethylene glycol (PEG) between about 100 and about 500 average molecular weight, (v) dimethyl sulfoxide (DMSO), and (vi) acetic acid.

7. A specimen jar containing at least the solution of claim 1.

8. The solution of claim 1 which further comprises (iv) polyethylene glycol (PEG) between about 100 and about 500 average molecular weight, (v) dimethyl sulfoxide (DMSO), and (vi) acetic acid.

9. The solution of claim 1, wherein said ketone is acetone.

10. The solution of claim 1, wherein said alcohol is isopropyl alcohol.

11. The solution of claim 1, wherein said ketone is acetone and said alcohol is isopropyl alcohol.

12. A non-aqueous solution, wherein the non-aqueous solution comprises (i) a ketone, (ii) an alcohol, and (iii) polyethylene glycol (PEG);

wherein the volume ratio of alcohol to ketone is from about 1;

1 to about 6;

1.

13. The solution of claim 12, wherein said PEG is between about 100 and about 500 average molecular weight.

14. The solution of claim 13 which further comprises (iv) dimethyl sulfoxide (DMSO).

15. The solution of claim 12, wherein the volume ratio between alcohol and ketone is from about 1:

1 to about 3;

1.

16. The solution of claim 12, wherein said ketone is acetone and said alcohol is isopropyl alcohol.

17. A method for processing tissue for histology, wherein the process comprises (a) hardening fresh, partially fixed, or fixed tissue in the solution of claim 12;

and (b) impregnating the hardened tissue in a wax solution.

18. The solution of claim 12, wherein said ketone is acetone.

19. The solution of claim 12, wherein said alcohol is isopropyl alcohol.

20. A specimen jar containing at least the solution of claim 12.