Method and apparatus for automated pre-treatment and processing of biological samples

• US 20060134793A1
• Filed: 07/07/2005
• Published: 06/22/2006
• Est. Priority Date: 07/23/2004
• Status: Active Grant

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Dako Denmark A/S (Agilent Technologies, Inc.)
• Inventors: Key, Marc E., Bland, Loren L., Devore, Scott A., Lovborg, Uffe
• Granted Patent: US 7,867,443 B2
• Time in Patent Office: Days
• US Class Current: 436/63
• 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 33/5306 Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• CPC Class Codes: Y10T 436/11 Automated chemical analysis
• CPC Class Codes: Y10T 436/111666 Utilizing a centrifuge or compartmented rotor
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

The present invention concerns a method and apparatus for automatic staining or other processing of a biological sample on a slide, perhaps robotically, by applying predetermined amounts of reagents in a predetermined sequence according to a processing protocol, the processing including pre-treatment steps, under the control of an adaptive processing control system. Further provided is a method of antigen retrieval, the method comprising contacting, within a heated processing tank of an automated staining apparatus, at least one biological sample with at least one composition for antigen retrieval comprising at least one chelating agent, at least one detergent and at least 50% by volume of at least one liquid that facilitates antigen retrieval.

Claims

1. A method for automated antigen retrieval and staining of a biological sample on a slide, comprising the steps of:

a.) providing an apparatus comprising a heatable processing tank for performing antigen retrieval upon the sample, a slide holder for holding the slide and a moveable probe for administering reagents to the sample during staining;

b.) mounting the slide onto the slide holder;

c.) causing the sample to contact a solution heated within the processing tank;

d.) removing the slide from the processing tank after a duration of time appropriate for antigen retrieval; and

e.) administering the reagents to the sample with the probe, wherein the solution comprises at least one chelating agent, at least one detergent and at least 50% by volume of at least one liquid that facilitates antigen retrieval.

2. The method of claim 1, wherein the at least one chelating agent is chosen from the group consisting of citric acid, sodium citrate and EDTA.

3. The method of claim 1, wherein the at least one detergent is chosen from Tween 20, NP40, Saponin, and Brij.

4. The method claim 1, wherein the at least one liquid that facilitates antigen retrieval is a glycol.

5. The method of claim 4, wherein the glycol is chosen from the group consisting of ethylene glycol and polypropylene glycol.

6. The method of claim 1, wherein the at least one liquid that facilitates antigen retrieval is a glycerol.

7. The method of claim 1, wherein the at least one liquid that facilitates antigen retrieval comprises a mixture between a first and a second liquid.

8. The method of claim 7, wherein the first liquid is propylene glycol.

9. The method of claim 7, wherein the second liquid is ethylene glycol.

10. The method of claim 1, wherein the boiling point of the solution is at least 120°

C. at sea level.

11. The method of claim 1, wherein the boiling point of the solution is at least 131°

C. at sea level.

12. The method of claim 1, wherein the viscosity of the solution is between 15-235 centipoise at 20°

C.

13. The method of claim 1, wherein the viscosity of the solution is between 1-10 centipoise at 20°

C.

14. The method of claim 1, wherein the processing tank is heated by a heating element disposed below the processing tank.

15. The method of claim 1, wherein the processing tank is heated by a heating blanket at least partially surrounding the processing tank.

16. The method of claim 14, wherein an insulation blanket at least partially surrounds the heating blanket.

17. The method of claim 1, wherein the slide holder is disposed within a slide rack housed within a drawer of the apparatus.

18. The method of claim 1, wherein the slide holder rotates the slide into a first orientation prior to the contact between the sample and the solution and rotates the selected slide into a second orientation prior to the administering of the reagents to the sample with the probe.

19. The method of claim 18, wherein the first orientation is substantially vertical and the second orientation is substantially horizontal.

20. The method of claim 1, wherein the probe is disposed upon a moveable arm.

21. The method of claim 1, wherein a second biological sample upon a second slide mounted onto a second slide holder within the apparatus is stained simultaneously with the staining of the sample.

22. The method of claim 21, wherein the second sample is caused to contact the solution.

23. A method for automated de-paraffinization, antigen retrieval and staining of a biological sample on a slide, comprising the steps of:

a.) providing an apparatus comprising a heatable processing tank for performing de-paraffinization and antigen retrieval upon the sample, a slide holder for holding the slide and a moveable probe for administering reagents to the sample during staining;

b.) mounting the slide onto the slide holder;

c.) causing the sample to contact a first solution within the processing tank, the first solution causing de-paraffinization of the sample;

d.) draining the first solution from the processing tank;

e.) causing the sample to contact a second solution heated within the processing tank;

f.) removing the slide from the processing tank after a duration of time appropriate for antigen retrieval; and

g.) administering the reagents to the sample with the probe,

wherein the second solution comprises at least one chelating agent, at least one detergent and at least 50% by volume of at least one liquid that facilitates antigen retrieval.

24. The method of claim 23, wherein the at least one chelating agent is chosen from the group consisting of citric acid, sodium citrate and EDTA.

25. The method of claim 23, wherein the at least one detergent is chosen from Tween 20, NP40, Saponin, and Brij.

26. The method claim 23, wherein the at least one liquid that facilitates antigen retrieval is a glycol.

27. The method of claim 26, wherein the glycol is chosen from the group consisting of ethylene glycol and polypropylene glycol.

28. The method claim 23, wherein the at least one liquid that facilitates antigen retrieval is a glycerol.

29. The method of claim 23, wherein the boiling point of the second solution is at least 120°

C. at sea level.

30. The method of claim 23, wherein the slide holder rotates the slide into a first orientation prior to the contact between the sample and the first solution and rotates the selected slide into a second orientation prior to the administering of the reagents to the sample with the probe.

31. The method of claim 23, wherein a second biological sample upon a second slide mounted onto a second slide holder within the apparatus is stained simultaneously with the staining of the sample.

32. The method of claim 31, wherein the second sample is caused to contact at least one member of the group consisting of the first solution and the second solution.