In situ heat induced antigen recovery and staining apparatus and method

• US 20060275861A1
• Filed: 05/24/2006
• Published: 12/07/2006
• Est. Priority Date: 07/08/1999
• Status: Active Grant

• Current Assignee: Lee H. Angros
• Original Assignee: Lee H. Angros
• Inventors: Byers, Thomas Lee, Angros, Lee
• Granted Patent: US 7,951,612 B2
• Time in Patent Office: Days
• US Class Current: 435/40.500
• CPC Class Codes: B01J 2219/0036 Nozzles
• CPC Class Codes: B01J 2219/00477 Means for pressurising the reaction vessels
• CPC Class Codes: B01J 2219/00495 Means for heating or cooling the reaction vessels
• CPC Class Codes: B01J 2219/00533 essentially rectangular
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01L 2200/04 Exchange or ejection of cartridges, containers or reservoirs
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/0822 Slides
• CPC Class Codes: B01L 2300/14 Means for pressure control
• CPC Class Codes: B01L 2400/0478 pistons
• CPC Class Codes: B01L 3/0293 for liquids
• CPC Class Codes: B01L 7/54 using spatial temperature gradients
• CPC Class Codes: G01N 1/312 for samples mounted on planar substrates
• CPC Class Codes: G01N 1/44 Sample treatment involving radiation, e.g. heat
• CPC Class Codes: G01N 33/52 Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72
• CPC Class Codes: G01N 33/521 Single-layer analytical elements
• CPC Class Codes: Y10T 436/11 Automated chemical analysis
• CPC Class Codes: Y10T 436/110833 Utilizing a moving indicator strip or tape
• CPC Class Codes: Y10T 436/112499 with sample on test slide
• CPC Class Codes: Y10T 436/114165 with step of insertion or removal from test line
• CPC Class Codes: Y10T 436/114998 : with treatment or replacement of aspirator element [e.g., cleaning, etc.]

Abstract

Contemplated herein is an automated microscope slide antigen recovery and staining apparatus and method that features a plurality of individually operable miniaturized pressurizable reaction compartments for individually and independently processing a plurality of individual microscope slides. The apparatus preferably features independently movable slide support elements each having an individually heatable heating plate. Each slide support element preferably supports a single microscope slide. Each microscope slide can be enclosed within an individual pressurizable reaction compartment. Pressures exceeding 1 atm or below 1 atm can be created and maintained in the reaction compartment prior to, during or after heating of the slide begins. Because of the ability to pressurize and regulate pressure within the reaction compartment, and to individually heat each slide, each slide and a liquid solution or reagent thereon can be heated to temperatures that could not be obtained without the enclosed pressurized environment of the reaction compartment. A reagent dispensing strip having a plurality of reconfigurable reagent modules may also be used.

Claims

1. A method of treating a microscope slide having a biological specimen thereon, comprising:

providing a plurality of independently operable reaction modules, each reaction module comprising;

a reaction compartment having an inner space;

a slide support element able to support a microscope slide in a horizontal position, the slide support element positionable within or adjacent the inner space of the reaction compartment for sealing the microscope slide therein;

a heating element for heating the microscope slide;

a reagent strip support device for supporting a reagent strip having a plurality of reagent containers each of which contains or is able to contain a reagent therein, wherein the reagent strip support device supports the reagent strip in a position external to and adjacent the reaction compartment; and

a dispensing element for engaging the reagent container thereby causing the reagent to be delivered from the reagent container into the inner space of the reaction compartment and onto the microscope slide;

and wherein each of the reaction compartments of the plurality of reaction modules is individually and independently pressurizable and wherein each of the heating elements of the slide support elements of the plurality of reaction modules is individually and independently heatable;

disposing the microscope slide onto the slide support element;

positioning the slide support element and microscope slide thereon within the reaction compartment;

placing the reagent strip onto the reagent strip support device;

actuating the dispensing element to cause the reagent to be delivered onto the microscope slide;

actuating the heating element to heat the slide; and

pressurizing the reaction compartment to maintain an internal pressure which exceeds atmospheric pressure or depressurizing the reaction compartment to a pressure less than atmospheric pressure.

2. The method of claim 1, wherein the step of pressurizing the reaction compartment occurs before, during, or after the heating of the microscope slide by the heating element.

3. The method of claim 1, wherein the reaction compartment has a cylindrical, tubular shape for enhancing pressure distribution within the reaction compartment.

4. The method of claim 1, wherein the slide support element of each reaction module is moved independently in relation to each other slide support element, the reagent strip of each reaction module is moved independently in relation to each other reagent strip, and the dispensing element of each reaction module is moved independently in relation to each other dispensing element.

5. The method of claim 1, wherein the reaction compartment is pressurized to a pressure of above 0 to 350 psig (101.3-2514 kPa), to a pressure of 1 to 100 psig (108.2-790.6 kPa), to a pressure of 5 to 50 psig (135.8-446.0 kPa), or to a pressure of 10 to 40 psig (170.3-377.0 kPa), or is depressurized as low as 100 kPa, 10 kPa, 1 kPa, 100 Pa, 10 Pa, 1 Pa or 0.1 Pa.

6. The method of claim 1, wherein the reagent disposed onto or about the microscope slide is heated to a temperature of 25°

C. to 37°

C., 37°

C. to 56°

C., 56°

C. to 85°

C., 85°

C. to 100°

C., 100°

C. to 125°

C., 125°

C. to 135°

C., 135°

C. to 150°

C., 150°

C. to 175°

C., 175°

C. to 200°

C., 200°

C. to 225°

C., 225°

C. to 250°

C., 250°

C. to 275°

C., 275°

C. to 300°

C., 300°

C. to 325°

C., to 325°

C. to 350°

C.

7. The method of claim 1 wherein the step of positioning the slide support element to enclose the microscope slide comprises moving the slide support element of the reaction module into the reaction compartment while the reaction compartment is stationary, or moving the reaction compartment while the slide support element is stationary, or moving the slide support element and moving the reaction compartment.

8. The method of claim 1 wherein the reagent strip is positioned in a dispensing position by moving the reagent strip support device thereby moving the reagent strip to the dispensing position, or by moving the reagent strip on the reagent strip support device while the reagent strip support device is stationary.

9. The method of claim 1 comprising the step of moving the reagent strip support device in either a forward or reverse direction to carry the reagent strip in either a forward or reverse direction, or maintaining the reagent strip support device stationary and moving the reagent strip thereon in either a forward or reverse direction.

10. The method of claim 1 wherein the reaction module comprises at least one of an air duct for pressurizing the reaction compartment or causing mixing of the reagent on the slide, a cooling duct for enhancing the rate of cooling of the heating element after heating, a supply port for delivering a liquid to the slide support element, and a drainage duct for removing reagents supplied to the microscope slide.

11. The method claim 1 comprising delivering reagent from the reagent strip into the reaction compartment via a reagent conduit in the reaction module.

12. The method claim 1 further comprising at least one of heating the reagent conduit for heating the reagent delivered therethrough, heating the reaction compartment, and heating the reagent strip or portions thereof using a heating device in the reagent strip support device.

13. The method of claim 1 comprising dispensing a solution in a cavity in the slide support element below the microscope slide and heating the solution in the cavity.

14. The method of claim 1 wherein the dispensing element is operable independently of the reagent strip support device, and wherein the dispensing element can dispense a reagent or solution from a reagent or solution source remote from the reagent strip, and wherein reagent can be applied from the reagent strip or reagent or solution from a remote source when the slide support element is disposed inside or outside of the reaction compartment, and wherein suction, or liquid, air, or gas under pressure can be applied to the microscope slide via the dispensing element to cause removal of a reagent or solution from the microscope slide.

15. The method of claim 1 comprising the step of tilting the slide support element within the reaction compartment or outside thereof to allow drainage of reagent or solution from the microscope slide.

16. A method of treating a microscope slide, comprising:

providing a plurality of independently operable reaction modules, each reaction module comprising;

a reaction compartment having an inner space;

a slide support element able to support a microscope slide in a horizontal position, the slide support element positionable within or adjacent the inner space of the reaction compartment for sealing the microscope slide therein; and

a dispensing element for dispensing a reagent into the reaction compartment;

disposing the microscope slide onto the slide support element;

positioning the microscope slide within the reaction compartment;

pressurizing the reaction compartment to maintain an internal pressure which exceeds atmospheric pressure; and

actuating the dispensing element to cause the reagent to be delivered into the reaction compartment while the reaction compartment is pressurized and wherein the reagent is delivered at a pressure which exceeds the pressure within the reaction compartment.

17. The method of claim 16 comprising heating the microscope slide and reagent within the reaction compartment.

18. A method of treating a microscope slide, comprising:

providing a plurality of independently operable reaction modules, each reaction module comprising;

a reaction compartment having an inner space;

a slide support element able to support a microscope slide in a horizontal position, the slide support element positionable within or adjacent the inner space of the reaction compartment for sealing the microscope slide therein; and

a dispensing element for dispensing a reagent into the reaction compartment;

disposing the microscope slide onto the slide support element;

positioning the microscope slide within the reaction compartment and enclosing the microscope slide therein;

actuating the dispensing element to cause the reagent to be delivered into the reaction compartment; and

pressurizing the reaction compartment to maintain an internal pressure which exceeds atmospheric pressure.

19. The method of claim 18 comprising heating the microscope slide and reagent within the reaction compartment.

20. A method of treating a microscope slide, comprising:

providing a reaction module comprising;

a reaction compartment having an inner space; and

a slide support element able to support a microscope slide in a horizontal position, the slide support element positionable within or adjacent the inner space of the reaction compartment for sealing the microscope slide therein;

disposing the microscope slide onto the slide support element;

positioning the slide support element and microscope slide thereon within the reaction compartment;

pressurizing the reaction compartment to maintain an internal pressure which exceeds atmospheric pressure; and

disposing a reagent onto the microscope slide while the reaction compartment is pressurized.

21. The method of claim 20 comprising the additional step of heating the microscope slide and the reagent thereon.

22. A method of treating a microscope slide, comprising:

providing a plurality of independently operable reaction modules, each reaction module comprising;

a reaction compartment having an inner space;

a slide support element able to support a microscope slide in a horizontal position, the slide support element positionable within or adjacent the inner space of the reaction compartment for sealing the microscope slide therein; and

a dispensing element for dispensing a reagent into the reaction compartment;

disposing the microscope slide onto the slide support element;

positioning the microscope slide within the reaction compartment;

depressurizing the reaction compartment to maintain an internal pressure which is less than atmospheric pressure; and

actuating the dispensing element to cause the reagent to be delivered into the reaction compartment while the reaction compartment is depressurized.