Apparatus for high quality, continuous throughput, tissue fixation-dehydration-fat removal-impregnation

• US 6,586,713 B2
• Filed: 12/15/2000
• Issued: 07/01/2003
• Est. Priority Date: 08/20/1997
• Status: Expired due to Term

• Current Assignee: Miami University, University of Miami
• Original Assignee: University of Miami
• Inventors: Morales, Azorides, Essenfeld, Ervin, Essenfeld, Harold
• Primary Examiner(s): Leung, Philip H.
• Application Number:
  US09/736,388
• Publication Number:
  US 20010000487A1
• Time in Patent Office: 928 Days
• Field of Search: 219/679, 219/700, 219/701, 219/726, 219/762, 342/59, 607/101, 607/102, 607/154, 435/173.1
• US Class Current: 219/679
• 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. An apparatus for continuous processing of tissue specimens comprising:

(a) a microwave processor unit which sequentially fixes, dehydrates, and defats said tissue specimens with a series of non-aqueous solutions in a vessel, and agitates and heats said vessel of the processor and its contents by microwave energy;

wherein sources for the non-aqueous solutions are fluidly coupled to said vessel of the processor to expose tissue specimens to said series of non-aqueous solutions and to drain said vessel of the processor; and

(b) an impregnator unit which sequentially impregnates said processed tissue specimens with a series of paraffin solutions in a vessel, and heats said vessel of the impregnator and its contents under less than atmospheric pressure during impregnation;

wherein processing of a later tissue specimen in said microwave processor unit is initiated before impregnation of an earlier tissue specimen in said impregnator unit has been completed.

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

(a) a microwave processor unit which sequentially fixes and dehydrates said tissue specimens with a series of non-aqueous solutions in a vessel, and heats said vessel of the processor and its contents by microwave energy;

wherein sources for the non-aqueous solutions are fluidly coupled to said vessel of the processor to expose tissue specimens to said series of non-aqueous solutions and to drain said vessel of the processor; and

(b) an impregnator unit which sequentially impregnates said processed tissue specimens with a series of paraffin solutions in a vessel, and heats said vessel of the impregnator and its contents under less than atmospheric pressure during impregnation;

wherein processing of a later tissue specimen in said microwave processor unit is initiated before impregnation of an earlier tissue specimen in said impregnator unit has been completed.

3. The apparatus of claim 2, wherein at least one of the series of non-aqueous solutions is comprised of an admixture of ketone and alcohol, wherein the volume ratio of alcohol to ketone is between about 1:

1 to about 3;

1.

4. The apparatus of claim 2, wherein at least one of the series of non-aqueous solutions is comprised of (a) 40% acetone, 40% isopropyl alcohol, and 20% low molecular weight polyethylene glycol by volume or (b) 10% acetone, 60% isopropyl alcohol, and 30% low molecular weight polyethylene glycol by volume;

wherein low molecular weight for a polymer is an average molecular weight of about 300.

5. The apparatus of claim 2, wherein at least one of the series of non-aqueous solutions is comprised of mineral oil and paraffin which has been degassed and dehydrated.

6. The apparatus of claim 2, wherein said tissue specimen is fixed, dehydrated and impregnated in a total time of about 65 minutes or less.

7. The apparatus of claim 2, wherein said tissue specimen is fixed, dehydrated and impregnated in a total time of less than about 90 minutes.

8. The apparatus of claim 2, wherein said tissue specimen is fixed, dehydrated and impregnated in a total time of less than about two hours.

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

(a) a microwave unit which hardens said tissue specimen with a non-aqueous solution in a first vessel, and heats with microwave energy and agitation said non-aqueous solution in the first vessel, wherein said non-aqueous solution in the first vessel is comprised of ketone and alcohol;

(b) a microwave unit which hardens said tissue specimen with a non-aqueous solution in a second vessel, and heats with microwave energy and agitation said non-aqueous solution in the second vessel, wherein said non-aqueous solution in the second vessel is comprised of less concentrated ketone and more concentrated alcohol as compared to said non-aqueous solution in the first vessel;

(c) a vacuum unit which impregnates said hardened tissue specimen with a wax solution in a third vessel, and heats under less than atmospheric pressure said non-aqueous solution in the third vessel, wherein said wax solution in the third vessel is comprised of paraffin wax; and

(d) a vacuum unit which impregnates said hardened tissue specimen with a wax solution in a fourth vessel, and heats under less than atmospheric pressure said non-aqueous solution in the fourth vessel, wherein said wax solution in the fourth vessel is comprised of more concentrated paraffin wax as compared to said wax solution in the third vessel;

wherein tissue specimens are sequentially transferred through the first, second, third and fourth vessels after processing for at least about 15 minutes in each vessel.

10. The apparatus of claim 9, wherein hardening a later tissue specimen in said first vessel is initiated before impregnation of an earlier tissue specimen in said fourth vessel has been completed.

11. The apparatus of claim 9, wherein said non-aqueous solution in the first vessel consists of about 40% acetone, about 40% isopropyl alcohol, and about 20% polyethylene glycol.

12. The apparatus of claim 9, wherein said non-aqueous solution in the second vessel consists of about 25% acetone, about 55% isopropyl alcohol, and about 20% mineral oil.

13. The apparatus of claim 9, wherein said wax solution in the third vessel consists of about 70% paraffin and about 30% mineral oil.

14. The apparatus of claim 9, wherein said wax solution in the fourth vessel consists of about 100% paraffin.

15. The apparatus of claim 9, wherein said tissue specimen is hardened and impregnated in a total time of about 65 minutes or less.

16. The apparatus of claim 9, wherein said tissue specimen is hardened and impregnated in a total time of less than about 90 minutes.

17. The apparatus of claim 9, wherein said tissue specimen is hardened and impregnated in a total time of less than about two hours.