ANTIGEN RECOVERY AND/OR STAINING APPARATUS AND METHOD

Description

[0001] The present invention is related to the field of treating samples on microscope slides and more specifically to the field of heat induced antigen recovery and staining.

[0002] Antigen recovery, also known as antigen unmasking, antigen epitope unmasking, antigen retrieval or heat induced epitope recovery (HIER) is a process in which biological samples (e.g., cells, tissues, blood, fluids) are treated under heat with a series of aqueous or non-aqueous reagents and buffers (e.g., citrate, EDTA, and urea) for the purpose of exposing the presence of specific types of antigens or biochemical features in the biological samples. HIER is regarded as a pre-treatment procedure to be performed prior to the beginning of a specific staining protocol to identify cellular components.

[0003] Biological samples must be preserved after removal from the body. This preservation process, known as fixation, kills and localizes the biological material. One of the most common fixatives used widely in the preservation of biological materials is formalin, a 10% aqueous solution of formaldehyde. This fixative, along with other widely utilized fixatives, produces a cross-linking network around specific sites in the biological material. These sites are known as antigens, and during the fixation process become "masked," by the fixative and thus "invisible" to detection by certain stains. HIER is used as a pre-treatment process to "unmask," "retrieve" or "recover." This process is usually conducted on formalin fixed paraffin embedded tissue sections or cellular preparations mounted on microscope slides.

[0004] U.S. Patent No. 5,244,787 teaches a process of antigen retrieval wherein one or more slides are placed in an aqueous solution within a microwave oven and heated to boiling or near-boiling temperatures. These slides are all treated together in a rack that has been placed in a bath of the solution. The slides are near boiling temperatures for 5-30 minutes, generally around 10 minutes. Due to excessive evaporation from the bath, the patent teaches that the solution should not drop below the biological sample on the slide because drying out of the sample is deleterious. This process further teaches that after boiling or near-boiling for several minutes, usually 5 minutes, one may have to add more solution to the container to prevent the solution from excessive evaporation and subsequent exposure of the samples on the slides. After the addition of more liquid, the process is continued until the desired time is completed. The disclosure of 5, 244, 787 is limited to the use of a microwave oven as the source of heating. More recent advances, which have been published, include the use of different types of heating devices such as electric pressure cookers, electric steamers, electric conduction heating surfaces utilizing pressure cookers, steamers, and also steam driving autoclaves (J. of Pathology, 179:347-352, 1996; Biotechnic & Histochemistry, 71(5):263-270, 1996; Biotechnic & Histochemistry, 71(4):190-195, 1996; J. of Histochemistry & Cytochemistry, 45 (3): 327-342, 1997).

[0005] Although these published methodologies treat the biological sample with different types of solutions and with varying types of chemicals and at different pH's, all teach that all slides are treated together in a bath of the heated solution. After the slides have cooled for a period of time, they are removed from the heating device and they are transferred to another apparatus where they are manually or automatically stained using various reagents. This pre-treatment process of heating and removing the slides from the heating device for staining in a separated apparatus is highly cumbersome and inefficient. The only automated HIER or antigen retrieval instrument available is the BIOGENEX i1000. This instrument, however, still employs the use of the known technology of treating the slides as a group in a container filled with heated solutions. A technician must still remove the slides from the antigen retrieval (heating) instrument and place them in an automated stainer instrument to complete the required staining protocol.

[0006] As noted herein, no currently available automated or semi-automated staining instruments specifically teach the ability to heat an aqueous or non-aqueous liquid for the unmasking of antigens. The instruments that do automated or semi-automated staining limit their scope to that task alone, and don't address the task of HIER or antigen retrieval pre-treatments. U.S. Patents 5,073,504 and 4,847,208 teach use of a chamber for enclosing and staining a microscope slide but neither teaches use of a heating device to boil a liquid and the user must add the primary antibody manually through a hinged door on top of the chamber. U.S. Patents 4, 777, 020; 4, 798, 706; and 4,801,431 teach use of a vertical staining "capillary gap" methodology wherein two special slides placed front to front causing an air gap through which liquids are drawn by capillary movement. This gap can only hold a small volume (approx. 300 microliters) of liquid. If heated to near boiling conditions the liquid would evaporate through all four open sides, immediately-causing the biological sample to dry. This end result is true also for another capillary gap instrument, shown in U.S. Patent 5,804,141. U.S. Patents 5,595,707; 5,654,200; 5,654,199, 5,595,707; and 5,650,327 teach reducing evaporative loss by utilizing an oil layer on top of the aqueous layer. This is somewhat effective in reducing the amount of evaporative loss at 37°C but the volume of the aqueous layer (approx. 300 microliters) is again minimal, and if heated to boiling, would cause the aqueous layer to dry out leaving only the oil layer present thus damaging the biological sample unless more aqueous reagent was applied during the treatment process. U.S. Patent 5,425,918 also teaches use of small amounts of liquids that are sprayed on the slide and can only heat the slide to 37°C. U.S. Patents 5,645,144 and 5,947,167 teach use of an open top chamber present around the slide and use a rotating cover above the slides to reduce evaporation. There is no teaching of high temperature heating of a liquid for a substantial amount of time. Further, even if one would increase the temperature of the slide, the loosely rotating top of the chamber would allow so much evaporative loss that the solution would never reach boiling or near boiling temperatures, nor would it maintain the boiling conditions for 10 minutes or longer. U.S. Patent 5, 645, 114 teaches use of small volumes of liquids (up to 500 microliters) and has no ability to stop evaporative loss if the slide temperature reaches boiling conditions.

[0007] U.S. Patent 5,654,200 discloses "Automated Slide Processing Apparatus With Fluid Injector". U.S. Patent 5,804,141 discloses "Reagent Strip Slide Treating Apparatus".

[0008] U.S. Patent 5,439,649 discloses "Automated Staining Apparatus". U.S. Patent 5,645,114 discloses "Dispensing Assembly With Interchangeable Cartridge Pumps". EP 0 402 994 discloses "Processing Apparatus For A Chemical Reaction Pack".

[0009] As a result, none of these systems could hold sufficient liquid on top of a slide (e.g., 4ml) and are enclosed in a chamber which is properly vented to minimize the energy loss from evaporation to cause sufficient heating to boil the liquid on the slide for the length of time generally required to cause antigen unmasking (e.g., 10-30 minutes).

[0010] There remains a need for an apparatus which can perform the task of HIER with subsequent staining treatment without the need of switching the slides from one apparatus to another and wherein the treatment of all microscope slides can occur simultaneously thereby increasing efficiency. Of the automated strainers available today, there is not one instrument that has the ability to overcome the inherent problems of heating an aqueous or non-aqueous solution at a sufficient volume without the undesirable effect of evaporative heat loss and subsequent volume decrease of the solution. The negative effects of evaporation are significant. The ability of a liquid to reach boiling or near boiling temperature on a microscope slide is dependent on the containment and control of the steam or vapor generated during the heating process. It is the object of the invention contemplated herein to provide a completely automated HIER apparatus which can recover antigens with multiple types of recovery buffers simultaneously, each specific to its respective microscope slide and which can also be used to stain the microscope slides as well.

[0011] In a first embodiment the present invention provides an in situ antigen recovery and staining apparatus for treating a biological sample on a microscope slide as claimed in claim 1 hereof.

[0012] In a second embodiment the subject invention provides a method of treating a microscope slide as claimed in claim 26 hereof.

DESCRIPTION OF THE DRAWINGS

[0013] 

Figure 1 is a perspective view of an apparatus of the invention (shown without a pressing element for crushing a reagent capsule).

Figure 2 is a cross-sectional view of the apparatus of Figure 1 (shown with a pressing element for crushing a reagent capsule).

Figure 3A is a cross-sectional view of the apparatus of Figure 1 (shown with a reaction compartment having a raised slide support surface) taken through line 3A-3A.

Figure 3B is a cross-sectional view of the apparatus of Figure 1 (shown with a reaction compartment having a lowered slide support surface) taken through line 3B-3B.

Figure 4 is a cross-sectional view of an alternative embodiment of the apparatus of the present invention having an alternate type of slide support surface.

Figure 5 is a perspective view of a reagent strip of the present invention.

Figure 6 is a cross-sectional view of the reagent strip of Figure 5 taken through the line 6-6.

Figure 7 is an elevational view of a modular apparatus containing a plurality of the apparatus of Figure 1.

Figure 8 is a flow chart showing a preferred sequence of steps in the method of the present invention.

DETAILED DESCRIPTION OF THE INVENTION

[0014] The present invention is disclosed in claims 1 and 28.

[0015] The present description discloses an automated method and apparatus for treating biological samples on microscope slides for unmasking ("retrieving" or "recovering") epitopes or antigens of the biological samples and then staining or otherwise treating the biological samples. The automated apparatus comprises an array of individual reaction compartments, each of which is used to treat a single microscope slide (also referred to herein as a "slide"), wherein each reaction compartment preferably can function and can be controlled independently of the other reaction compartments in the array. Each reaction compartment in the array comprises a support element comprising a surface upon which a microscope slide can be supported and positioned adjacent or inserted into the compartment for treatment with a reagent. The support element further comprises, in a preferred embodiment, a conduction type heating element for heating the microscope slide to a predetermined treatment temperature when desired. The support element with the microscope slide thereon can be raised into or adjacent the reaction compartment for treatment of the microscope slide, or lowered or removed from the reaction compartment for placement of a microscope slide onto or removed from the support surface or for removal of a reagent or rinsing solution from the microscope slide during the treatment process.

[0016] Reagents, such as antibodies, enzymes, rinse buffers, antigen recovery buffers, or stains, are contained in an individualized reagent dispensing strip which is specific for each microscope slide to be treated. Since each microscope slide and reaction compartment is generally provided with its own reagent dispensing strip, each microscope slide can be treated independently with a different set of reagents (a particular treatment protocol) while being treated simultaneously with other microscope slides. Similarly, each microscope slide can be heated separately, as well as treated with a different treatment protocol. The apparatus of the present disclosure therefore comprises a plurality of individualized reaction compartments in a chamber which can be substantially closed for minimizing evaporation during heating. A microscope slide can be supported in each reaction compartment, and each microscope slide can be heated separately therein. A reagent dispensing strip containing a plurality of individually contained reagents (reagent "bubbles", "blisters" or "capsules") is positioned upon an upper portion of each reaction compartment, and at an appropriate time, a reagent from each reagent dispensing strip is expelled from a reagent capsule under compression and is thereby applied to the biological sample on the microscope slide. Or, a reagent, such as an antigen recovery buffer can be introduced via a separate dispenser. The term "reagent" is defined herein to include any type of fluid material that may be applied to the biological material on the microscope slide, including antibodies, stains, enzymes, buffers, rinses, or washes, or any other material applied in the process of antigen recovery or treating the biological material on the microscope slide to be viewed under the microscope.

[0017] During an antigen recovery step, the microscope slide, sample, and antigen recovery buffer thereon are heated to an appropriate temperature for a predetermined duration to cause the antigen recovery buffer to react with the sample on the microscope slide, after which the antigen recovery buffer is removed from the microscope slide, preferably by washing or flooding the microscope slide or chamber containing the microscope slide with a rinse buffer and allowing the rinse buffer to drain off by gravity or by blowing the solution off the microscope slide using pressurized air. Each microscope slide may be treated in the same manner, or may be treated with different reagents using a different treatment protocol, preferably simultaneously, yet independently.

[0018] When a reagent is provided via a reagent dispensing strip, the apparatus is preferably equipped with a drive mechanism for causing the reagent dispensing strip to be advanced in a forward direction wherein each reagent capsule in succession is positioned above an aperture in the compartment through which the reagent in the capsule is delivered. The reagent dispensing strip may be advanced using rollers positioned along the upper end of the compartment or a pushing mechanism which pushes upon the rear end of the reagent dispensing strip. The reagent in the reagent capsule of the reagent dispensing strip is to be applied to the microscope slide by a pressing mechanism which, in a preferred version, compresses and thereby crushes the reagent capsule and causes the reagent to be expelled and deposited directly onto the microscope slide.

[0019] In a method of the present description, a plurality of microscope slides, each having thereon a sample to be treated, is provided. Each microscope slide is positioned upon a support element which is then moved into an application position. A plurality of reagent dispensing strips is provided, one for each microscope slide to be treated. Each microscope slide is subjected to an antigen recovery step then is treated by applying a reagent from its corresponding reagent dispensing strip. Each microscope slide can be handled differently, if desired, during the treatment cycle. After a predetermined duration, the microscope slide and support element is moved to a removal position wherein the reagent is removed, preferably in between reagent applications, by treatment with a rinsing solution to remove the reagent prior to further treatment. Each microscope slide can be treated according to the treatment protocol specific to that sample or that particular microscope slide. All microscope slides may be treated using the same protocol, or one or more, or all, of the microscope slides may be treated using a different protocol.

[0020] An example of a treatment protocol comprises:

1) antigen recovery, approximately 10 minutes at approximately 98°C,

2) cool, approximately 20 minutes,

3) rinse buffer,

4) primary antibody, approximately 30 minutes,

5) rinse,

6) biotinylated linking antibody, approximately 10 minutes,

7) rinse buffer,

8) peroxidase labeled streptavidin label,

9) rinse buffer,

10) 3,3'-diaminobenzidine chromogen,

11) rinse buffer,

12) chromogen enhancer,

13) rinse buffer, and

14) counter stain.

[0021] A variety of other treatment protocols are well known to those of ordinary skill in the art and further discussion of them herein is not deemed necessary. Each microscope slide, if necessary, may be heated prior to application of the reagent, if necessary, then may be cooled as the reagent is removed, then reheated, if necessary, prior to or after addition of the next reagent. The entire process is run automatically once the microscope slide is disposed onto the support element, and the reagent dispensing strip is positioned upon the upper side of the reaction compartment.

[0022] Turning now to the drawings, a specific embodiment of the apparatus of the present invention is shown in Figures 1-6. Although Figures 1-6 show a preferred version of the invention, it will be understood that the embodiment shown in Figures 1-6 is but one of many possible versions of the apparatus enabled herein which will come to the mind of a person of ordinary skill in the art.

[0023] Shown in Figure 1, and designated therein by the general reference numeral 10 is an antigen recovery and staining apparatus constructed in accordance with the present invention. The antigen recovery and staining apparatus 10 comprises a treatment chamber 12 which further comprises a plurality of reaction compartments 14 (see Figures 2 - 4). Preferably the treatment chamber 12 generally comprises from about 10 to about 20 reaction compartments 14 but may contain more or fewer. Each reaction compartment 14, when enclosed, minimizes evaporation of a reagent solution when a microscope slide is exposed to high temperature pretreatment conditions. Each reaction compartment 14 has an upper side 16 having an opening 18 therein, a lower side 20, and a pair of sidewalls 22 which extend from the rear end 23a of the treatment chamber 12 to the front end 23b of the treatment chamber 12. Positioned above each reaction compartment 14 is a reagent dispensing strip holder 24 for holding and guiding a reagent dispensing strip 26 (see Figures 5 and 6). Each reagent dispensing strip 26 has a front end 28 and a rear end 30 and a plurality of capsules 32 made of a crushable plastic material such as polyethylene or another suitable material (e.g., polypropylene or polystyrene) and which may include one or more multiple capsules 32a. The size of each capsule 32 or multiple capsule 32a may be adjusted to accommodate the amount of reagent which is desired to be applied to a microscope slide 44. Each capsule 32 or multiple capsule 32a contains a reagent or treatment solution which is intended to be applied to a biological material on the microscope slide 44. Multiple capsule 32a is useful in a method wherein two or more reagents must be contained separately before being applied to the microscope slide 44. When the multiple capsule 32a is crushed by the pressing mechanism 36, two or more reagents contained within the capsule 32a are combined and simultaneously applied to the microscope slide 44.

[0024] Other embodiments of the reagent dispensing strip 26 and the reagent capsule 32 and multiple capsule 32a will readily be apparent to one of ordinarily skill in the art. For example, each reagent dispensing strip 26 may comprise a one or more "blank" spaces for insertion of individualized capsules 32 by a user. Below each capsule 32 or multiple capsule 32a is an aperture or weak area 34 in the reagent dispensing strip 26 through which the reagent in the capsule 32 or multiple capsules 32 can be forced by a pressing mechanism 36. The "blank" space or space left by the puncturing of a capsule 32 or 32a, or vents in the reagent dispensing strip 26 may function to release pressure, steam or vapors produced during the treatment process. The reagent dispensing strip 26 is advanced in a direction 37 toward the front end 23b of the treatment chamber 12 by a reagent strip drive mechanism 38 driven, for example, by an electric motor which in Figures 1. 3A and 3B is shown as a pushing mechanism comprising a threaded shaft, but which may instead by a mechanism (not shown) comprising rollers which drive, draw or "pull" the reagent strip holder 24 in a forward direction 37.

[0025] Each reaction compartment 14 further comprises at its lower side 20 a slide support assembly 39 comprising a plurality of slide support elements 40 each having a slide tray 42 upon which the microscope slide 44 can be positioned and held for treatment. With the microscope slide 44 disposed on the slide support element 40, the slide support element 40 and the microscope slide 44 are positioned in an application position to fit adjacent the lower side 20 of the reaction compartment 14, thereby constituting an openable bottom of the reaction compartment 14. The slide support element 40 further has a heating element 46 incorporated therein for heating the microscope slide 44 as discussed elsewhere herein. In one embodiment, the slide support element 40 has a hinge 48 for enabling the slide support element 40 to be moved (raised) into an application position (Figure 3A) and therefrom lowered into an opened position (see Figure 3B). Alternatively, the slide support element 40 may be raised and lowered into position by another mechanism, such as a stepper motor 58 and screw drive 59 mechanism (Figure 4). Each reaction compartment 14 further comprises a manifold 50 which comprises, in a preferred embodiment, a plurality of reagent dispensing ports or elements including, for example but not limited to , an antigen recovery buffer dispenser 51 connected via an antigen recovery buffer supply line 51a to an antigen recovery buffer supply (not shown), a rinse buffer dispenser 52 connected via a rinse buffer supply line 52a to a rinse buffer supply (not shown) and an air pressure nozzle 54 connected via an air line 54a to an air supply (not shown). The antigen recovery buffer dispenser 51 applies an antigen recovery buffer to the microscope slide 44 for the antigen recovery treatment step prior to staining or other preparation of the biological material on the microscope slide 44. The rinse buffer dispenser 52 applies a rinse buffer 56 to the microscope slide 44 to rinse a reagent from the microscope slide 44. The air pressure nozzle 54 functions to clear away a rinse buffer 56 from the microscope slide 44. Dispensers 51 and 52 may be used to dispense other reagents, and may constitute more than, or fewer than, the dispensers shown in Figures 2, 3A, 3B, and 4. The microscope slide 44 is generally disposed in a removal position for facilitating removal of the rinse buffer 56 as shown in Figures 1 and 3B. Each slide support element 40, in a preferred embodiment, can be heated or moved independently of any other slide support element 40, although one of ordinary skill in the art can envision that the slide support elements 40 may be designed to operate in concert, i.e., simultaneously.

[0026] The antigen recovery and staining apparatus 10 can be controlled automatically wherein predetermined sequences and operations are carried out using various electromechanical systems which are not shown but which are well known to those of ordinary skill in the art. For example, each of the steps of raising into a treatment position and lowering into a removal position each of the slide support elements 40, applying an antigen recovery buffer, advancing each reagent dispensing strip 26, compressing each capsule 32 or 32a of the reagent dispensing strip 26, heating each microscope slide 44 on the slide support surface 40, applying a rinse buffer 56 to the microscope slide 44, removing the rinse buffer 56 or other reagent from the microscope slide 44, and treating each microscope slide 44 independently can be automatically controlled and programed using programming methods and devices well known in the art. Because each reaction compartment 14 and slide support element 40 can be controlled independently, a microscope slide 44 can even be removed or inserted even while other reaction compartments 14 are in operation.

[0027] Preferably, a microprocessor, not shown, controls the antigen recovery and staining apparatus 10. That is, an operator programs the microprocessor with information such as which reaction compartments 14 are to be used and to what temperature each is to be heated and at which steps, then programs the particular treatment protocol to be performed on the sample on each microscope slide 44 on each slide support element 40. variables in these protocols can include the particular type of reagent dispensing strip 26 to be used, the time that each reagent or treatment solution on the reagent dispensing strip 26 will be allowed to react with the sample on the microscope slide 44, whether the microscope slide 44 will be heated, and if so to what temperature and for how long, and the manner in which the microscope slide 44 will be rinsed, for example. Other variables not listed herein may also be programmed.

[0028] The apparatus of the present description may further comprise a modular apparatus 60 comprising a plurality of antigen recovery and staining apparatuses 10 each serving as an individual module in the modular apparatus 60. The individual modules can be "stacked" together for example, as shown in Figure 7, or may be oriented in any other desirable manner.

[0029] Shown in Figure 8 is a schematic drawing which describes the preferred method of the present invention. In the first step, a microscope slide 44 which has a sample disposed thereon is provided, and is disposed onto a slide support element 40 which is moved into an application or treatment position adjacent or against the reaction compartment 14. If a plurality of microscope slides 44 are supplied, each microscope slide 44 is disposed on a separate microscope slide support element 40 and the microscope slides 44 are moved independently or simultaneously into an application position.

[0030] Once in the application position, an antigen recovery buffer is initially applied to the sample on the microscope slide 44. Microscope slide 44 is then heated to a desired, predetermined temperature, for example from about 140°C to about 160°C whereby the antigen recovery buffer is heated to a temperature of from about 90°C to about 100°C, for example. The microscope slide 44 is allowed to react with the reagent for a predetermined length of time, for example, about 10 to about 30 minutes, preferably at about 95° to about 98°C. Venting of steam may occur through small holes (not shown) in the reagent strip 26 or elsewhere in the reaction compartment 14.

[0031] It is not necessary to add additional antigen recovery buffer during this step. After the reaction period is over, the slide support element 40 and the microscope slide 44 thereon are moved (lowered or dropped) to a removal position, if necessary, where the antigen recovery buffer is removed from the microscope slide 44, for example, by applying a rinsing solution or buffer to the microscope slide 44 or by gravity or by pressurized air. A rinse solution or buffer may be applied and removed more than once for treatment or for removal of a particular reagent before or after lowering the microscope slide 44 to the removal position. It may be desirable to add rinse buffer to the microscope slide 44 to cool the microscope slide 44 prior to lowering the microscope slide 44 to the removal position, for example, by adding rinse buffer 56 to the antigen recovery buffer before the microscope slide 44 is moved to the application position. After the microscope slide 44 has been treated for antigen recovery, another reagent can then be applied for treatment of the sample on the microscope slide 44. In this step, the microscope slide 44 and slide support element 40 are then returned to the application position, a reagent is applied, and is then removed after the treatment period. The series of steps may be repeated. When the treatment of the sample is completed, the microscope slide 44 is removed from the slide support element 40 for further treatment or analysis apart from the antigen recovery and staining apparatus 10.

[0032] Changes may be made in the construction and the operation of the various components, elements and assemblies described herein or in the steps or the sequence of steps of the methods described herein without departing from the scope of the invention as defined in the following claims.

[0033] The invention illustratively disclosed herein suitably may be practiced in the absence of any element which is not specifically disclosed herein.

[0034] The following claims are entitled to the broadest possible scope consistent with this application. The claims shall not necessarily be limited to the preferred embodiments or to the embodiments shown in the examples

Claims

1. An in situ antigen recovery and staining apparatus (10) for treating a biological sample on a microscope slide (44) having a reagent disposed thereon, comprising:

a slide support assembly (39) comprising a plurality of movable slide support elements (40) each for supporting a single microscope slide (44);

a heating element (46) incorporated into or adjacent each slide support element (40) for
heating the single microscope slide (44) on the slide support element (40), wherein each microscope slide (44) and the reagent thereon can be independently heated; and

a plurality of individual reaction compartments (14) each having an inner space into
which the microscope slide (44) can be positioned, characterized in that:

each reaction compartment (14) is capable of being substantially closed during heating of
the microscope slide (44) therein, and each reaction compartment when substantially closed can be vented to release pressure, steam or vapors during the heating of the microscope slide (44), and wherein each slide support element (40) having the microscope slide (44) thereon is independently movable relative to each other slide support element (40) having a microscope slide (44) thereon such that each microscope slide (44) on each slide support element (40) can be automatically independently treated using a different treatment protocol; and wherein each microscope slide (44) can be removed from or inserted into the corresponding reaction compartment (14) while other reaction compartments (14) are in operation.

2. The apparatus (10) of claim 1 wherein the reaction compartment (14) comprises a small holes to release vapor pressure.

3. The apparatus (10) of claims either of claims 1 or 2 wherein the reagent on the microscope slide (44) is supplied from a reagent dispensing element (26), (51), (52).

4. The apparatus (10) of claim 3 wherein each reagent dispensing element (26), (51), (52) is operable independently of and/or simultaneously with each other reagent dispensing element (26), (51), (52).

5. The apparatus (10) of either claims 3 or 4 wherein the reagent dispensing element (26), (51), (52) is a reagent dispensing strip (26) comprising at least one individually contained reagent (32).

6. The apparatus (10) of any one of claims 1-5 further comprising a port (51), (52) or dispenser (51), (52) for independently and/or simultaneously providing an additional reagent to the microscope slide (44).

7. The apparatus (10) of any one of claims 1-6 wherein each separate heating element (46) can operate simultaneously to heat each microscope slide (44) disposed on the corresponding slide support element (40).

8. The apparatus (10) of any one of claims 5-7 wherein each reagent dispensing strip (26) contains a plurality of the individually contained reagents (32) positioned upon an upper portion of each reaction compartment from which a reagent is expelled under compression and is thereby applied to the biological sample on the microscope slide.

9. The apparatus of any one of claims 1-8 wherein the microscope slide (44) can be heated such that the reagent disposed thereon can attain a temperature of from about 90°C to about 100°C.

10. The apparatus of any one of claims 1-9 wherein the microscope slide (44) can be heated to a temperature of 140°C to 160°C.

11. The apparatus of any of claims 1-10 wherein each slide support element (40) and microscope slide (44) thereon can be moved to a removal position for removing the reagent from each microscope slide (44) and wherein each slide support element (40) and microscope slide (44) thereon can be returned to an application position.

12. The apparatus of any one of claims 1-11 wherein each slide support element (40) is automatically simultaneously movable relative to each other slide support element (40).

13. The apparatus (10) of any one claims 1-12 comprising means (54) for removing the reagent from the microscope slide (44) via pressurized air, vacuum, or aspiration, and wherein the means (54) for removing the reagent can operate independently and/or simultaneously for each microscope slide (44).

14. The apparatus (10) of any one of claims 1-13 comprising means (52) for rinsing the microscope slide (44) with a rinse solution.

15. The apparatus (10) of any one of claims 1-14 wherein the reagent can be removed from each microscope slide (44) independently and/or simultaneously with a rinse solution.

16. The apparatus (10) of any one of claims 1-15 wherein the microscope slide (44) can be heated to a temperature adequate to induce antigen recovery of the biological sample on the microscope slide (44).

17. The apparatus (10) of any one of claims 1-16 wherein additional antigen recovery buffer does not need to be added to the microscope slide (44) while the microscope slide (44) is being heated during an antigen recovery treatment protocol.

18. The apparatus (10) of any one of claims 5-17 comprising a pressing mechanism (36) for dispensing the at least one individually contained reagent (32) from the reagent dispensing strip (26) and wherein each pressing mechanism (36) is able to function independently of each other pressing mechanism (36).

19. The apparatus (10) of any one of claims 1-18 comprising a microprocessor for controlling operations within the apparatus (10).

20. The apparatus of any one of claims 1-19 comprising an air pressure nozzle (54) for simultaneously and/or independently applying a stream of pressurized air to the reagent on each microscope slide (44).

21. The apparatus of any one of claims 5-20 wherein each reagent strip (26) is automatically simultaneously and independently movable relative to each other reagent dispensing strip (26).

22. A method of treating a microscope slide (44), comprising:

providing an apparatus (10) comprising a plurality of movable slide support elements (40) each for supporting a single microscope slide (44), and each slide support element (40) having a heating element (46) incorporated therein or adjacent thereto for heating the single microscope slide (44) on the slide support element (40), wherein each microscope slide (44) and the reagent thereon can be independently heated by the heating element (46);

the apparatus further comprising a plurality of individual reaction compartments (14)
each having an inner space into which the single microscope slide (44) can be positioned for treatment, the method characterized in that:

in the apparatus (10), each reaction compartment (14) is capable of being substantially closed during heating of the microscope slide (44), and wherein the reaction compartment (14) when substantially closed can be vented to release pressure, steam or vapors during the heating of the microscope slide (44), and wherein each slide support element (40) having the microscope slide (44) thereon is independently movable relative to each other slide support element (40) having a microscope slide (44) thereon during operation of the apparatus (10) such that each microscope slide (44) on each slide support element (40) can be automatically independently treated;

providing a plurality of the microscope slides (44), each having a treatment sample disposed thereon;

disposing each microscope slide (44) on the slide support element (40) corresponding thereto;

treating each microscope slide (44) by applying a reagent thereon wherein the reagent can be applied independently to each microscope slide (44); and

wherein each microscope slide (44) can be removed from or inserted into the corresponding reaction compartment (14) while other reaction compartments (14) are in operation.

23. The method of claim 22 wherein the reagent is applied from a reagent dispensing strip (26) which corresponds to only a single microscope slide (44).

24. The method of claim 22 or 23 wherein each microscope slide (44) is placed within the reaction compartment (14) corresponding thereto.

25. The method of either of claims 23 or 24 wherein each reagent dispensing strip (26) can operate independently of each other reagent dispensing strip (26).

26. The method of either of any one of claims 22-25 wherein in the step of removing the reagent, the reagent is removed by pressurized air from each microscope slide (44) independently and/or simultaneously.

27. The method of any one of claims 22-25 wherein in the step of removing the reagent, the reagent is removed by vacuum or aspiration from each microscope slide (44) independently and/or simultaneously.

28. The method of any one of claims 22-27 wherein the step of removing the reagent from each microscope slide (44) includes independently and/or simultaneously rinsing the microscope slide (44) with a rinse solution.

29. The method of any one of claims 22-28 comprising the additional step of heating at least one of said microscope slides (44) using the separate heating element (46).

30. The method of claim 29 wherein said microscope slide (44) is heated to a temperature of at least 140°C to 160°C.

31. The method of either of any one of claims 29-30 wherein the microscope slide (44) is heated such that the reagent is heated to a temperature of between about 90°C-100°C.

32. The method of any one of claims 22-31 wherein the reagent on the microscope slide (44) is heated to a temperature of about 98°C for about 10 minutes.

33. The method of any one of claims 22-32 wherein each microscope slide (44) is heated to a temperature adequate to induce antigen recovery of the treatment sample on the microscope slide (44).

34. The method of any one of claims 22-33 comprising the step of moving each slide support element (40) and microscope slide (44) thereon to a removal position for removing the reagent from each microscope slide (44) thereby treated or for removing the microsocpe slide (44) from the slide support element (40) and comprising the step of moving each slide support element (40) and microscope slide (44) thereon to a treatment position for applying the reagent thereto.

35. The method of any one of claims 22-34 wherein the reagent is one selected from the group of an antigen recovery buffer, antibody, enzyme, buffer, rinse, wash, label, chromogen enhancer, and stain.

36. The method of claim 35 wherein additional antigen recovery buffer is not added while the microscope slide (44) is being heated during an antigen recovery treatment protocol. 37 22-36

37. The method of any one of claims 22-36 wherein at least one of the microscope slides (44) of the plurality of microscope slides (44) is treated using a treatment protocol which is different from that used for at least one other microscope slide (44) of the plurality of microscope slides.

38. The method of any one of claims 22-37 wherein an additional reagent is dispensed from a port or dispenser (51), (52) which is operable independently of and/or simultaneously with each of other port or dispenser (51), (52).

39. The method of any one of claims 22-38 wherein the reaction compartment is vented via small holes to release vapor pressure.

40. The method of any one of claims 22-39 wherein each slide support element (40) is automatically simultaneously movable relative to each other slide support element (40).

41. The method of any one of claims 23 or 25 wherein each reagent dispensing strip (26) is independently movable relative to each other reagent dispensing strip (26).

Ansprüche

1. Vorrichtung (10) zum Nachweisen und Anfärben von Antigenen in situ zur Behandlung einer biologischen Probe auf einem Objektträger (44), auf dem sich ein Reagens befindet, wobei die Vorrichtung folgendes umfasst:

eine Objektträger-Trägereinheit (39), die eine Mehrzahl beweglicher Objektträger-Trägerelemente (40) umfasst, die jeweils zum Tragen eines einzelnen Objektträgers (44) dienen;

ein Heizelement (46), das in jedem Objektträger-Trägerelement (40) oder neben diesem inkorporiert ist, um den einzelnen Objektträger (44) an dem Objektträger-Trägerelement (40) zu erhitzen, wobei jeder Objektträger (44) und das Reagens darauf unabhängig voneinander erhitzt werden können; und

eine Mehrzahl einzelner Reaktionskammern (14), die jeweils einen Innenraum aufweisen, in dem der Objektträger (44) positioniert werden kann, dadurch gekennzeichnet, dass:

jede Reaktionskammer (14) im Wesentlichen geschlossen werden kann, während der Objektträger (44) darin erhitzt wird, und wobei jede Reaktionskammer, wenn sie im Wesentlichen geschlossen ist, entlüftet werden kann, um Druck, Dampf oder Dunst freizusetzen, während der Objektträger (44) erhitzt wird, und wobei jedes Objektträger-Trägerelement (40), welches den Objektträger (44) daran aufweist, unabhängig beweglich ist im Verhältnis zu jedem anderen Objektträger-Trägerelement (40), welches einen Objektträger (44) daran aufweist, so dass jeder Objektträger (44) an jedem Objektträger-Trägerelement (40) automatisch unter Verwendung eines anderen Behandlungsprotokolls unabhängig behandelt werden kann; und wobei jeder Objektträger (44) aus der entsprechenden
Reaktionskammer (14) entfernt oder in diese eingeführt werden kann, während sich andere Reaktionskammern (14) im Betrieb befinden.

2. Vorrichtung (10) nach Anspruch 1, wobei die Reaktionskammer (14) kleine Öffnungen zur Freisetzung von Dampfdruck umfasst.

3. Vorrichtung (10) nach Anspruch 1 oder 2, wobei das Reagens auf dem Objektträger (44) durch ein Reagensabgabeelement (26), (51), (52) zugeführt wird.

4. Vorrichtung (10) nach Anspruch 3, wobei jedes Reagensabgabeelement (26), (51), (52) unabhängig von und/oder gleichzeitig mit jedem anderen Reagensabgabeelement (26), (51), (52) funktionsfähig ist.

5. Vorrichtung (10) nach Anspruch 3 oder 4, wobei es sich bei dem Reagensabgabeelement (26), (51), (52) um einen Reagensabgabestreifen (26) handelt, der mindestens ein abgeschlossenes Reagens (32) umfasst.

6. Vorrichtung (10) nach einem der Ansprüche 1 bis 5, wobei diese ferner einen Anschluss (51), (52) oder eine Abgabeeinrichtung (51), (52) zur unabhängigen und/oder gleichzeitigen Bereitstellung eines zusätzlichen Reagens an den Objektträger (44) umfasst.

7. Vorrichtung (10) nach einem der Ansprüche 1 bis 6, wobei jedes separate Heizelement (46) gleichzeitig arbeiten kann, um jeden Objektträger (44) zu erhitzen, der sich auf dem entsprechenden Objektträger-Trägerelement (40) befindet.

8. Vorrichtung (10) nach einem der Ansprüche 5 bis 7, wobei jeder Reagensabgabestreifen (26) eine Mehrzahl einzeln abgeschlossener Reagenzien (32) aufweist, die auf einem oberen Abschnitt jeder Reaktionskammer positioniert sind, von dem ein Reagens unter Druck ausgestoßen wird und dadurch der biologischen Probe auf dem Objektträger zugeführt wird.

9. Vorrichtung nach einem der Ansprüche 1 bis 8, wobei der Objektträger (44) so erhitzt werden kann, dass das sich darauf befindende Reagens eine Temperatur zwischen etwa 90 °C und etwa 100 °C annehmen kann.

10. Vorrichtung nach einem der Ansprüche 1 bis 9, wobei der Objektträger (44) auf eine Temperatur zwischen 140 °C und 160 °C erhitzt werden kann.

11. Vorrichtung nach einem der Ansprüche 1 bis 10, wobei jedes Objektträgerelement (40) und der sich darauf befindende Objektträger (44) an eine Entfernungsposition bewegt werden können, um das Reagens von jedem Objektträger (44) zu entfernen, und wobei jedes Objektträger-Trägerelement (40) und der Objektträger (44) darauf an eine Anwendungsposition zurückgeführt werden können.

12. Vorrichtung nach einem der Ansprüche 1 bis 11, wobei jedes Objektträger-Trägerelement (40) automatisch gleichzeitig im Verhältnis zu jedem anderen Objektträger-Trägerelement (40) beweglich ist.

13. Vorrichtung (10) nach einem der Ansprüche 1 bis 12, wobei diese eine Einrichtung (54) zum Entfernen des Reagens von dem Objektträger (44) mittels Druckluft, Vakuum oder Ansaugen umfasst, und wobei die Einrichtung (54) zum Entfernen des Reagens für jeden Objektträger (44) unabhängig und/oder gleichzeitig arbeiten kann.

14. Vorrichtung (10) nach einem der Ansprüche 1 bis 13, wobei diese eine Einrichtung (52) zum Spülen des Objektträgers (44) mit einer Spüllösung umfasst.

15. Vorrichtung (10) nach einem der Ansprüche 1 bis 14, wobei das Reagens von jedem Objektträger (44) unabhängig und/oder gleichzeitig mit einer Spüllösung entfernt werden kann.

16. Vorrichtung (10) nach einem der Ansprüche 1 bis 15, wobei der Objektträger (44) auf eine Temperatur erhitzt werden kann, die dazu geeignet ist, einen Antigennachweis der biologischen Probe auf dem Objektträger (44) zu induzieren.

17. Vorrichtung (10) nach einem der Ansprüche 1 bis 16, wobei dem Objektträger (44) kein zusätzlicher Antigennachweispuffer hinzugefügt werden muss, während der Objektträger (44) während dem Antigenrücknachweisbehandlungsprotokoll erhitzt wird.

18. Vorrichtung (10) nach einem der Ansprüche 5 bis 17, wobei diese einen Pressmechanismus (36) zur Abgabe des mindestens einen einzeln abgeschlossenen Reagens (32) von dem Reagensabgabestreifen (26) umfasst, und wobei jeder Pressmechanismus (36) unabhängig von jedem anderen Pressmechanismus (36) arbeiten kann.

19. Vorrichtung (10) nach einem der Ansprüche 1 bis 18, wobei diese einen Mikroprozessor zur Steuerung der Operationen in der Vorrichtung (10) umfasst.

20. Vorrichtung nach einem der Ansprüche 1 bis 19, wobei diese eine Luftdruckdüse (54) für die gleichzeitige und/oder unabhängige Zufuhr eines Druckluftstroms an das Reagens auf jedem Objektträger (44) umfasst.

21. Vorrichtung nach einem der Ansprüche 5 bis 20, wobei jeder Reagensstreifen (26) automatisch gleichzeitig und unabhängig im Verhältnis zu jedem anderen Reagensabgabestreifen (26) beweglich ist.

22. Verfahren zur Behandlung eines Objektträgers (44), wobei das Verfahren folgendes umfasst:

das Bereitstellen einer Vorrichtung (10), die eine Mehrzahl beweglicher Objektträger-Trägerelemente (40) umfasst, die jeweils zum Tragen eines einzelnen Objektträgers (44) dienen, und wobei jedes Objektträger-Trägerelement (40) ein Heizelement (46) aufweist, das darin oder neben diesem inkorporiert ist, um den einzelnen Objektträger (44) an dem Objektträger-Trägerelement (40) zu erhitzen, wobei jeder Objektträger (44) und das Reagens darauf unabhängig voneinander durch das Heizelement (46) erhitzt werden können; und

wobei die Vorrichtung ferner eine Mehrzahl einzelner Reaktionskammern (14) umfasst, die jeweils einen Innenraum aufweisen, in dem der einzelne Objektträger (44) für eine Behandlung positioniert werden kann, wobei das Verfahren dadurch gekennzeichnet ist, dass:

in der Vorrichtung (10) jede Reaktionskammer (14) im Wesentlichen geschlossen werden kann, während der Objektträger (44) darin erhitzt wird, und wobei die Reaktionskammer (14), wenn sie im Wesentlichen geschlossen ist, entlüftet werden kann, um Druck, Dampf oder Dunst freizusetzen, während der Objektträger (44) erhitzt wird, und wobei jedes Objektträger-Trägerelement (40), welches den Objektträger (44) daran aufweist, unabhängig beweglich ist im Verhältnis zu jedem anderen Objektträger-Trägerelement (40), welches einen Objektträger (44) daran aufweist, so dass jeder Objektträger (44) an jedem Objektträger-Trägerelement (40) automatisch unabhängig behandelt werden kann;

das Bereitstellen einer Mehrzahl von Objektträgern (44), die jeweils eine darauf angeordnete Behandlungsprobe aufweisen;

das Anordnen jedes Objektträgers (44) auf dem entsprechenden Objektträger-Trägerelement (40);

das Behandeln jedes Objektträgers (44) durch Auftragen eines Reagens darauf, wobei das Reagens unabhängig auf jeden Objektträger (44) aufgetragen werden kann; und

wobei jeder Objektträger (44) aus der entsprechenden Reaktionskammer (14) entfernt oder in diese eingeführt werden kann, während sich andere Reaktionskammern (14) im Betrieb befinden.

23. Verfahren nach Anspruch 22, wobei das Reagens von einem Reagensabgabestreifen (26) zugeführt wird, der einem einzigen Objektträger (44) entspricht.

24. Verfahren nach Anspruch 22 oder 23, wobei jeder Objektträger (44) in der entsprechenden Reaktionskammer (14) platziert wird.

25. Verfahren nach Anspruch 23 oder 24, wobei jeder Reagensabgabestreifen (26) unabhängig von allen anderen Reagensabgabestreifen (26) funktionsfähig ist.

26. Verfahren nach einem der Ansprüche 22 bis 25, wobei in dem Schritt des Entfernens des Reagens das Reagens durch Druckluft unabhängig und/oder gleichzeitig von jedem Objektträger (44) entfernt wird.

27. Verfahren nach einem der Ansprüche 22 bis 25, wobei in dem Schritt des Entfernens des Reagens das Reagens unabhängig und/oder gleichzeitig durch Vakuum oder Ansaugen von jedem Objektträger (44) entfernt wird.

28. Verfahren nach einem der Ansprüche 22 bis 27, wobei der Schritt des Entfernens des Reagens von jedem Objektträger (44) das unabhängige und/oder gleichzeitige Spülen des Objektträgers (44) mit einer Spüllösung aufweist.

29. Verfahren nach einem der Ansprüche 22 bis 28, wobei dieses den zusätzlichen Schritt des Erhitzens mindestens eines der genannten Objektträger (44) unter Verwendung des separaten Heizelements (46) umfasst.

30. Verfahren nach Anspruch 29, wobei der genannte Objektträger (44) auf eine Temperatur von mindestens 140 °C bis 160 °C erhitzt wird.

31. Verfahren nach einem der Ansprüche 29 bis 30, wobei der Objektträger (44) so erhitzt wird, dass das Reagens auf eine Temperatur zwischen etwa 90 °C und 100 °C erhitzt wird.

32. Verfahren nach einem der Ansprüche 22 bis 31, wobei das Reagens auf dem Objektträger (44) etwa zehn Minuten lang auf eine Temperatur von etwa 98 °C erhitzt wird.

33. Verfahren nach einem der Ansprüche 22 bis 32, wobei jeder Objektträger (44) auf eine Temperatur erhitzt wird, die geeignet ist, um einen Antigennachweis der Behandlungsprobe auf dem Objektträger (44) zu induzieren.

34. Verfahren nach einem der Ansprüche 22 bis 33, wobei dieses den Schritt des Bewegens jedes Objektträger-Trägerelements (40) und eines Objektträgers (44) darauf an eine Entfernungsposition zur Entfernung des dadurch behandelten Reagens von jedem Objektträger (44) umfasst oder zum Entfernen des Objektträgers (44) von dem Objektträger-Trägerelement (40), und wobei das Verfahren den Schritt des Bewegens jedes Objektträger-Trägerelements (40) und jedes Objektträgers (44) darauf zum Auftragen des Reagens darauf an eine Behandlungsposition umfasst.

35. Verfahren nach einem der Ansprüche 22 bis 34, wobei das Reagens aus der Gruppe ausgewählt wird, die einen Antigennachweispuffer, einen Antikörper, ein Enzym, einen Puffer, eine Spüllösung, eine Waschlösung, einen Marker, einen Chromogenverstärker und einen Farbstoff umfasst.

36. Verfahren nach Anspruch 35, wobei kein zusätzlicher Antigennachweispuffer hinzugefügt wird, während der Objektträger (44) während einem Antigennachweisbehandlungsprotokoll erhitzt wird.

37. Verfahren nach einem der Ansprüche 22 bis 36, wobei mindestens einer der Objektträger (44) der Mehrzahl von Objektträgern (44) unter Verwendung eines Behandlungsprotokolls behandelt wird, das sich von dem Behandlungsprotokoll unterscheidet, das für mindestens einen anderen Objektträger (44) der Mehrzahl von Objektträgern verwendet wird.

38. Verfahren nach einem der Ansprüche 22 bis 37, wobei ein zusätzliches Reagens von einem Anschluss oder einer Ausgabeeinrichtung (51), (52) ausgegeben werden, der bzw. die unabhängig von und/oder gleichzeitig zu jedem anderen Anschluss oder jeder anderen Ausgabeeinrichtung (51), (52) betrieben werden kann.

39. Verfahren nach einem der Ansprüche 22 bis 38, wobei die Reaktionskammer über kleine Öffnungen zur Freisetzung von Dampfdruck entlüftet wird.

40. Verfahren nach einem der Ansprüche 22 bis 39, wobei jedes Objektträger-Trägerelement (40) automatisch gleichzeitig im Verhältnis zu jedem anderen Objektträger-Trägerelement (40) beweglich ist.

41. Verfahren nach einem der Ansprüche 23 oder 25, wobei jeder Reagensabgabestreifen (26) unabhängig beweglich ist im Verhältnis zu jedem anderen Reagensabgabestreifen (26).

Revendications

1. Appareil de démasquage et de coloration d'antigènes in situ (10) pour traiter un échantillon biologique sur une lame de microscope (44) sur laquelle a été disposé un réactif, comprenant :

un ensemble formant support de lame (39) comprenant une pluralité d'éléments de support de lame mobiles (40) chacun servant à supporter une lame de microscope unique (44) ;

un élément de chauffage (46) incorporé dans ou à proximité de chaque élément de support de lame (40) pour chauffer la lame de microscope unique (44) sur l'élément de support de lame (40), dans lequel chaque lame de microscope (44) et le réactif se trouvant dessus peuvent être chauffés indépendamment ; et

une pluralité de compartiments de réaction individuels (14) ayant chacun un espace interne dans lequel la lame de microscope (44) peut être positionnée, caractérisé en ce que :

chaque compartiment de réaction (14) peut être sensiblement fermé pendant le chauffage de la lame de microscope (44) située à l'intérieur, et chaque compartiment de réaction lorsqu'il est sensiblement fermé peut être ventilé pour relâcher de la pression, de la vapeur ou des vapeurs pendant le chauffage de la lame de microscope (44), et dans lequel chaque élément de support de lame (40) sur lequel se trouve la lame de microscope (44) peut être bougé de façon indépendante par rapport à un autre élément de support de lame (40) sur lequel se trouve une lame de microscope (44) de telle sorte que chaque lame de microscope (44) sur chaque élément de support de lame (40) peut être automatiquement traitée indépendamment au moyen d'un protocole de traitement différent ; et dans lequel chaque lame de microscope (44) peut être retirée de ou insérée dans le compartiment de réaction (14) correspondant tandis que d'autres compartiments de réaction (14) sont en fonctionnement.

2. Appareil (10) selon la revendication 1, dans lequel le compartiment de réaction (14) comprend de petits trous pour libérer la pression de vapeur.

3. Appareil (10) selon la revendication 1 ou 2, dans lequel le réactif sur la lame de microscope (44) est fourni à partir d'un élément de distribution de réactif (26), (51), (52).

4. Appareil (10) selon la revendication 3, dans lequel chaque élément de distribution du réactif (26), (51), (52) peut fonctionner indépendamment de et/ou simultanément à un autre élément de distribution de réactif (26), (51), (52).

5. Appareil (10) selon la revendication 3 ou 4, dans lequel l'élément de distribution de réactif (26), (51), (52) est une bande de distribution de réactif (26) comprenant au moins un réactif contenu individuellement (32).

6. Appareil (10) selon l'une quelconque des revendications 1 à 5, comprenant en outre un orifice (51), (52) ou un distributeur (51), (52) pour fournir de façon indépendante et/ou simultanée un réactif supplémentaire à la lame de microscope (44).

7. Appareil (10) selon l'une quelconque des revendications 1 à 6, dans lequel chaque élément de chauffage (46) séparé peut fonctionner simultanément pour chauffer chaque lame de microscope (44) disposée sur l'élément de support de lame (40) correspondant.

8. Appareil (10) selon l'une quelconque des revendications 5 à 7, dans lequel chaque bande de distribution de réactif (26) contient une pluralité de réactifs contenus individuellement (32) positionnés sur une partie supérieure de chaque compartiment de réaction à partir de laquelle un réactif est expulsé sous compression et est ainsi appliqué à l'échantillon biologique sur la lame de microscope.

9. Appareil selon l'une quelconque des revendications 1 à 8, dans lequel la lame de microscope (44) peut être chauffée de telle sorte que le réactif disposé sur celle-ci peut atteindre une température d'environ 90 °C à environ 100 °C.

10. Appareil selon l'une quelconque des revendications 1 à 9, dans lequel la lame de microscope (44) peut être chauffée à une température comprise entre 140 °C et 160 °C.

11. Appareil selon l'une quelconque des revendications 1 à 10, dans lequel chaque élément de support de lame (40) et lame de microscope (44) se trouvant dessus peuvent être déplacés à une position de retrait pour retirer le réactif de chaque lame de microscope (44) et dans lequel chaque élément de support de lame (40) et lame de microscope (44) se trouvant dessus peuvent être remis à une position d'application.

12. Appareil selon l'une quelconque des revendications 1 à 11, dans lequel chaque élément de support de lame (40) peut être automatiquement et simultanément déplacé par rapport à un autre élément de support de lame (40).

13. Appareil (10) selon l'une quelconque revendications 1 à 12 comprenant des moyens (54) pour retirer le réactif de la lame de microscope (44) par l'intermédiaire d'air sous pression, de vide ou d'aspiration, et dans lequel les moyens (54) pour retirer le réactif peuvent fonctionner de manière indépendante et/ou simultanée pour chaque lame de microscope (44).

14. Appareil (10) selon l'une quelconque des revendications 1 à 13 comprenant des moyens (52) pour rincer la lame de microscope (44) avec une solution de rinçage.

15. Appareil (10) selon l'une quelconque des revendications 1 à 14 dans lequel le réactif peut être retiré de chaque lame de microscope (44) de manière indépendante et/ou simultanée avec une solution de rinçage.

16. Appareil (10) selon l'une quelconque des revendications 1 à 15, dans lequel la lame de microscope (44) peut être chauffée à une température suffisante pour induire un démasquage d'antigène de l'échantillon biologique sur la lame de microscope (44).

17. Appareil (10) selon l'une quelconque des revendications 1 à 16, dans lequel un tampon de démasquage d'antigène supplémentaire n'a pas besoin d'être ajouté à la lame de microscope (44) alors que la lame de microscope (44) est chauffée pendant un protocole de traitement de démasquage d'antigène.

18. Appareil (10) selon l'une quelconque des revendications 5 à 17, comprenant un mécanisme de pression (36) pour distribuer l'au moins un réactif contenu individuellement (32) à partir de la bande de distribution de réactif (26) et dans lequel chaque mécanisme de pression (36) pour fonctionner indépendamment de chaque autre mécanisme de pression (36).

19. Appareil (10) selon l'une quelconque des revendications 1 à 18, comprenant un microprocesseur pour commander les opérations au sein de l'appareil (10).

20. Appareil selon l'une quelconque des revendications 1 à 19, comprenant une buse de pression d'air (54) pour appliquer simultanément et/ou indépendamment un flux d'air sous pression au réactif sur chaque lame de microscope (44).

21. Appareil selon l'une quelconque des revendications 5 à 20, dans lequel chaque bande de réactif (26) peut être automatiquement déplacée, de manière simultanée et indépendante, par rapport à chaque autre bande de distribution de réactif (26).

22. Procédé de traitement d'une lame de microscope (44) comprenant les étapes consistant à :

fournir un appareil (10) comprenant une pluralité d'éléments de support de lame mobiles (40) chacun étant destiné à supporter une lame de microscope unique (44), et chaque élément de support de lame (40) ayant un élément chauffant (46) incorporé dans celui-ci ou adjacent à celui-ci pour chauffer la lame de microscope unique (44) sur l'élément de support de lame (40), dans lequel chaque lame de microscope (44) et le réactif se trouvant dessus peuvent être chauffés indépendamment par l'élément chauffant (46) ;

l'appareil comprenant en outre une pluralité de compartiments de réaction individuels (14) ayant chacun un espace interne dans lequel la lame de microscope unique (44) peut être positionnée à des fins de traitement, le procédé étant caractérisé en ce que :

dans l'appareil (10), chaque compartiment de réaction (14) peut être sensiblement fermé pendant le chauffage de la lame de microscope (44), et dans lequel le compartiment de réaction (14) lorsqu'il est sensiblement fermé peut être ventilé pour relâcher de la pression, de la vapeur ou des vapeurs pendant le chauffage de la lame de microscope (44), et dans lequel chaque élément de support de lame (40) sur lequel se trouve la lame de microscope (44) peut être déplacé de façon indépendante par rapport à un autre élément de support de lame (40) sur lequel se trouve une lame de microscope (44) pendant le fonctionnement de l'appareil (10) de telle sorte que chaque lame de microscope (44) sur chaque élément de support de lame (40) peut être automatiquement traitée de façon indépendante ;

fournir une pluralité de lames de microscope (44), un échantillon de traitement étant disposé sur chacune d'elles ;

disposer chaque lame de microscope (44) sur l'élément de support de lame (40) qui lui correspond ;

traiter chaque lame de microscope (44) par l'application d'un réactif dessus, dans lequel le réactif peut être appliqué indépendamment sur chaque lame de microscope (44) ; et

dans lequel chaque lame de microscope (44) peut être retirée de ou insérée dans le compartiment de réaction (14) correspondant alors que d'autres compartiments de réaction (14) sont en fonctionnement.

23. Procédé selon la revendication 22, dans lequel le réactif est appliqué à partir d'une bande de distribution de réactif (26) qui correspond à une lame de microscope unique (44).

24. Procédé selon la revendication 22 ou 23, dans lequel chaque lame de microscope (44) est placée à l'intérieur du compartiment de réaction (14) qui lui correspond.

25. Procédé selon la revendication 23 ou 24, dans lequel chaque bande de distribution de réactif (26) peut fonctionner indépendamment d'une autre bande de distribution de réactif (26).

26. Procédé selon l'une quelconque des revendications 22 à 25 dans lequel, à l'étape d'élimination du réactif, le réactif est éliminé par de l'air sous pression de chaque lame de microscope (44) de manière indépendante et/ou simultanée.

27. Procédé selon l'une quelconque des revendications 22 à 25 dans lequel, à l'étape d'élimination du réactif, le réactif est éliminé par vide ou aspiration à partir de chaque lame de microscope (44) de manière indépendante et/ou simultanée.

28. Procédé selon l'une quelconque des revendications 22 à 27, dans lequel l'étape d'élimination du réactif à partir de chaque lame de microscope (44) comprend l'étape consistant à rincer de façon indépendante et/ou simultanée la lame de microscope (44) avec une solution de rinçage.

29. Procédé selon l'une quelconque des revendications 22 à 28 comprenant l'étape supplémentaire consistant à chauffer au moins l'une desdites lames de microscope (44) au moyen de l'élément de chauffage séparé (46).

30. Procédé selon la revendication 29, dans lequel ladite lame de microscope (44) est chauffée à une température au moins comprise entre 140 °C et 160 C.

31. Procédé selon la revendication 29 ou 30, dans lequel la lame de microscope (44) est chauffée de telle sorte que le réactif est chauffé à une température comprise entre environ 90 °C et 100 °C.

32. Procédé selon l'une quelconque des revendications 22 à 31, dans lequel le réactif sur la lame de microscope (44) est chauffé à une température d'environ 98 °C pendant environ 10 minutes.

33. Procédé selon l'une quelconque des revendications 22 à 32, dans lequel la lame de microscope (44) est chauffée à une température appropriée pour induire un démasquage d'antigène de l'échantillon biologique sur la lame de microscope (44).

34. Procédé selon l'une quelconque des revendications 22 à 33, comprenant l'étape consistant à déplacer chaque élément de support de lame (40) et lame de microscope (44) se trouvant dessus à une position de retrait pour retirer le réactif de chaque lame de microscope (44) ainsi traitée ou pour retirer la lame de microscope (44) de l'élément de support de lame (40) et comprenant l'étape consistant à déplacer chaque élément de support de lame (40) et lame de microscope (44) se trouvant dessus à une position de traitement pour y appliquer le réactif.

35. Procédé selon l'une quelconque des revendications 22 à 34, dans lequel le réactif est choisi dans le groupe constitué par un tampon de démasquage d'antigène, un anticorps, une enzyme, un tampon, un rinçage, un lavage, une étiquette, un amplificateur chromogène, et une coloration.

36. Procédé selon la revendication 35, dans lequel un tampon de démasquage d'antigène supplémentaire n'est pas ajouté pendant que la lame de microscope (44) est chauffée pendant un protocole de traitement de démasquage d'antigène.

37. Procédé selon l'une quelconque des revendications 22 à 36, dans lequel au moins une des lames de microscope (44) de la pluralité de lames de microscope (44) est traitée au moyen d'un protocole de traitement qui est différent de celui utilisé pour au moins une autre lame de microscope (44) de la pluralité de lames de microscope.

38. Procédé selon l'une quelconque des revendications 22 à 37, dans lequel un réactif supplémentaire est distribué à partir d'un orifice ou d'un distributeur (51), (52) qui peut fonctionner indépendamment de et/ou simultanément à un autre orifice ou distributeur (51), (52).

39. Procédé selon l'une quelconque des revendications 22 à 38, dans lequel le compartiment de réaction est ventilé par de petits trous pour libérer la pression de vapeur.

40. Procédé selon l'une quelconque des revendications 22 à 39, dans lequel chaque élément de support de lame (40) est automatiquement déplacé de manière simultanée par rapport à un autre élément de support de lame (40).

41. Procédé selon l'une la revendication 23 ou 25, dans lequel chaque bande de distribution de réactif (26) est indépendamment mobile par rapport à chaque autre bande de distribution de réactif (26).

Drawing