Automated high volume slide staining system

ES 2 425 689 T3

DESCRIPTION

Automated slide staining system with high production speed

The present invention relates to medical diagnostic equipment. The invention has particular utility in connection with automated staining of biological samples on microscope slide plates, and will be described in connection with this use, although other uses are envisaged.

Many tissues do not retain enough color after processing to make their components visible under a bright field microscope. Accordingly, it is a common technique to add a color or colorants to fabrics for coloring. Hematoxylin and eosin ("H and E") stain is probably the most widely used histological stain. Its popularity is based on its relative simplicity and ability to clearly reveal a huge number of different tissue structures. Hematoxylin can be prepared in multiple ways and has wide applicability to tissues from different locations. Essentially, the hematoxylin component colors the cell nuclei blue / black with satisfactory intranuclear detail, while the eosin colors the cell cytoplasm and most connective tissue fibers with different shades and intensities of pink, orange and red. .

An accurate diagnosis depends on the examination by a pathologist or cytologist of colored microscope slides, usually H and E paraffin sections, with H and E staining in bulk having been carried out by an automatic staining machine. The need for consistency is vital to avoid difficult histological interpretation. In general, automated coloring apparatuses allow accurate and uniform coloring, consistent differentiation, and dehydration by adjusting the time of each stage.

Some standard procedures are routinely applicable to tissue staining on slide plates. First, paraffin sections must be deparaffinized, because most of the stains are applied in aqueous or alcoholic solutions that would not penetrate paraffin-infiltrated tissues. After the slide plates have been dewaxed, the slide plates are usually subjected to an oven or other heated chamber to remove dewaxed solvent and to adhere the tissues to the plates. Tissues can be stained using, for example, standard stains such as hematoxylin and eosin. Finally, the coating is accomplished by adhering a thin glass plate to the colored tissue section and then sealing it with a mounting agent, thereby creating a hermetically sealed environment suitable for filing. So far, all these steps have been carried out manually by a histology technician, a profession that is disappearing among laboratory technicians dedicated to the technique of heat treatment and staining of human tissue samples for their reading and interpretation by a pathologist. .

US 6,296,809 B1 discloses apparatus and methods for automatically staining or treating multiple tissue samples mounted on microscope slide plates. The individualized control of the temperature of the slide plates is achieved by means of a heating system that has thermal platforms mounted radially on the carousel for heating the slide plates and detecting the temperature of each one of them. The heating system also allows, if necessary, the automated removal of waxes.

WO 01/42796 A1 discloses a method and apparatus for molecular definition on a large scale and, with large throughput of tissue samples, collecting a tissue sample from a donor among a set of donor samples, placing a sample of the donor specimen at a designated location on a recipient device, providing substantial copies of the set, performing a separate biological analysis on each copy, and storing the analysis results. The results can be compared to determine if there are correlations or discrepancies between the results of different biological analyzes in each assigned location and also making the comparison with the clinical information of the human patient from which the tissues were obtained. Similar analysis results on corresponding sections of the assembly can be used as quality control devices, for example by subjecting the devices to a single, simultaneous investigative process. The uniform interpretation of the sets can be obtained, and can be compared with interpretations of different observers.

US 6,017,495 discloses a coloring apparatus, for coloring tissue samples, placed on microscope slide plates, comprising a series of coloring stations and other work stations, in which the coloring stations receive containers containing liquid baths to receive cuvettes containing microscope plates with the topical samples, and a transport mechanism having a lifting device arranged to move over the containers and to place or collect trays from the containers, and transfer the trays between workstations in accordance with a program controlled coloring process. Containers are transported and positioned in a series of horizontal rows positioned one above the other on a stepped base such as a rostrum, the rostrum having a gradient that allows visual inspection of individual containers at work stations from one viewing location. ahead of the device.

ES 2 425 689 T3

Therefore, according to one aspect, it is a problem to provide a method for automatically preparing tissue samples on microscope slide plates for pathological analysis and an automated slide processing apparatus that facilitates heat treatment and drying. and at the same time prevents cross contamination of the slide plates during dewaxing and staining.

This problem is solved by the method having the features disclosed in claim 1 and in the automated slide plate processing apparatus having the features of claim 15. Preferred embodiments are defined in the dependent claims.

By supporting all the slide plates in the same plane, heat treatment and drying is facilitated and cross-contamination of the slide plates during dewaxing and staining is also prevented.

The present invention provides an automatic slide staining system for applying stains to sections of biological tissues mounted on microscope slide plates. More particularly, the present invention provides an automated apparatus for deparaffinizing, staining, and protecting a series of slide plates carrying biological samples, in a fully automated and integrated system. In a preferred embodiment, the present invention provides an automated apparatus comprising a series of stacked workstations, in which specimen-bearing slide plates can be dried, heat-treated, dewaxed, and prepared for staining, applying stains. and sealing or coating the plates so that they can be stored for future analysis and study or as a permanent record of the analyzes carried out, and a transport elevator for moving a slide tray carrying a series of tissue sample plates between the series of workstations. In another embodiment, the invention is directed to a method for the automatic preparation of tissue samples on microscope slide plates for pathological analysis, comprising heat treating the tissue sample on the slide plate by causing the instrument to apply heat to the tissue. sufficiently to adhere to the slide plate; dewaxing the tissue sample by contact with it with a dewaxing fluid at a temperature above the melting point of the paraffin and then washing the liquefied paraffin by dragging it, coloring the tissue sample by contact with a coloring reagent and protecting the slide plate with a coating by contacting the colored tissue sample on the plate with a previously glued coating and an adhesive-activating fluid.

Other features and advantages of the present invention will be appreciated from the following detailed description of the invention, taken in conjunction with the following drawings, in which:

Figure 1 is a simplified plan view, and Figures 1A and 1B are partial front and rear perspective views of a high speed automated coloring apparatus made in accordance with a first embodiment of the present invention;

Figure 1C is a perspective view from the inside, and Figure 1D is a perspective view with parts disassembled from the outside of the tray storage station or "garage" part of the present invention;

Figure 2 is a perspective view and Figure 2A an exploded view showing details of a support tray for a tissue sample carrier plate used in the present invention;

Figure 3 is an exploded perspective view of a portion of a dewaxing module of a first embodiment of the present invention;

Figure 3A is a top plan view of the nozzle and manifold portion of the dewaxing module of Figure 3;

Figure 4 is a view, similar to Figure 3, of the heating or heat treatment module of a first embodiment of the present invention;

Figure 5 is a perspective view showing detail of the dewaxing fluid recovery system of a first embodiment of the present invention;

Figure 6 is a schematic line diagram of the dewaxing fluid system of a first embodiment of the present invention;

Figure 7 is a view, similar to Figure 3, of a coloring applicator station module of the present invention;

Figure 8 is a perspective view of the coating module of the present invention;

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Figure 8A is a perspective view of a coating cartridge portion of the coating module shown in Figure 8;

Figure 9 is an exploded view showing details of the carrier plate tray transport and the elevator portion of the present invention;

Figures 10A and 10B show two different perspective views showing details of the XY table of the tray transport of the carrier plates and the elevator portion of the present invention;

Fig. 11 is a flow chart showing a first embodiment of the present invention; Y

Figure 12 is a view, similar to Figure 1, of a second embodiment of an automated high speed coloring apparatus, made in accordance with the present invention;

Fig. 13 is an exploded view of a combination heat treating and dewaxing module part of the second embodiment of the present invention; Y

FIG. 14 is a view, similar to FIG. 11, of a flow chart showing the second embodiment of the present invention.

The staining system of the present invention carries out all stages of the processing, staining and coating of biological samples mounted on microscope slides efficiently and at high speed. More particularly, the slide plates carrying the biological samples are placed on a slide plate tray, and the plate tray carrying the sample slide plates are loaded into the system according to the present invention, so that the slide plates They are guided through a sequence of steps in which the slide plates are thermally prepared, dewaxed, colored, and finally coated. A method of the present invention is directed to a method for the automatic preparation of tissue samples on microscope slide plates for pathological analysis that comprises, heat treatment of the tissue sample on the slide plate by causing the instrument to apply heat to the tissue sufficiently to adhere to the slide, dewaxing the tissue sample by contact with a dewaxing fluid at a temperature above the boiling point of the paraffin, and subsequent pouring of the liquefied paraffin by removing it; staining the tissue sample by contacting it with a staining reagent and coating the slide plate by contacting the stained tissue sample on the slide with a pre-glued coating and an adhesive activating fluid.

Referring to figure 1 of the drawings, the apparatus 20 of the first embodiment of the invention functions as a component or module of a system 22. The system 22 also comprises bulk fluid containers 32, 34, 36 and the corresponding equipment .

Referring also to Figures 1A and 1B, the apparatus 20 comprises a frame 40 supporting a stack of workstations comprising, for example, one or more modules or stations 120 for drying or heat treatment, waxing station or dewaxing or module 140, one or more coloring stations or modules 160 and a coating station or module 180 arranged in a tower 50. A lift and transport mechanism 52 is disposed adjacent tower 50 to transport a slide plate tray 54 designed to support a series of individual slide plates 56 that support tissue samples from a tray storage station or "garage" 60 via drying, waxing, coloring and coating operations.

Referring in particular to Figures 1B and 1C, the garage or storage station 60 comprises a pair of uprights 62A, 62B carrying a series of vertically spaced shelves 64 for receiving tray 54 of carrier plates. Referring also to Figure 1D, the garage or tray storage station 60 comprises a pivotally mounted door that facilitates access to a first shelf position (for clarity, the outer shell or garage lid 60 has been omitted). A tray drive assembly, indicated generally at 68, includes a pair of rotatably mounted drive wheels 70, driven by a drive motor and transmission 72, being disposed below the first shelf position. to move a tray in and out of gantry 66.

Referring in particular to Figures 2 and 2A, the slide plate tray 54 preferably comprises a slide plate tray or support 80 having a generally rectangular shape in plan, including a bottom wall 82, opposite side walls 84 and opposite end walls 86. The slide tray is typically formed by conventional injection molding using synthetic polymers intended for this use, which are well known in the art.

Tray 80 comprises a shelf 90 for supporting tissue sample plates to support slide plates in a substantially horizontal position in the same plane. Keeping all the plates

ES 2 425 689 T3 slides in the same plane facilitates drying, as will be described later, and also prevents cross-contamination of the slide plates during dewaxing and staining, as will be described later. The shelf 90 comprises a series of resilient slide plate supports 92 that limit axial, lateral, and vertical movement of the tissue sample slide plates 56 once positioned on the plate tray. The shelf 90 is supported above the bottom 80 of the tray at a sufficient height to prevent or reduce the formation of films or bubbles between the bottom of the tissue specimen slide and the bottom of the tray. Elastic slide supports 92 hold the individual specimen slides in position by exerting force on opposing edges 96 of the specimen slides. The bottom of the pannier tray slopes toward the middle to facilitate drainage to a central location 104 for evacuation of wax and stain removal fluids, as will be described in detail below. Tray 40 allows automated handling of specimen slides through the dewax drying, staining and coating steps. In a preferred embodiment, tray 80 includes splash guides 106 and is arranged to receive 16 sample slides positioned on a generally horizontal rack having a width of two plates and a height of eight.

In the embodiment shown, the coloring system comprises a drying / heat treatment station or module 120, a dewaxing station or module 40, a coloring station or module 160 and a coating station or module 180 arranged vertically in a tower 50 and controlled by a computer.

Referring to FIG. 4, a heat treating / drying station 120 comprises a thermally insulated department into which heat is supplied in a controlled manner for drying the slide samples. The drying / heat treating station 120 preferably comprises a modular unit and includes a convection heater 122, arranged to direct a flow of hot air over the surface of the sample slides. A feature and advantage of the present invention, which results from the horizontal presentation of the slides, is that convection drying is particularly effective.

Referring in particular to Figures 3 and 3A, dewaxing station 140 comprises a modular compartment and includes one or a series of single-use wash nozzles 142 directed downwardly at an angle relative to the sample slides. . Preferably, dewaxing station 140 comprises two arrays 144A, B of ten nozzles 142, each fed via common manifolds 146A, b with a suitable dewaxing fluid from a supply of dewaxing fluid 32, which in a preferred embodiment of the invention comprises hot water and detergent. Alternatively, a pair of nozzles may be mounted on a mobile device, and advanced from one pair of slides to another pair of slides.

Various dewaxing agents can be used and preferably comprise an aqueous fluid, such as that disclosed in US 6,855,559 B1 filed February 15, 2005, and US 6,544. 798, filed April 8, 2003, which include deionized water, citrate buffer (pH 6.0 - 8.0), Tris-HCl buffer (PH 6 - 10), phosphate buffer (pH 6.0 - 8.0 ), FSC buffer, APK wash^TM, buffers or acid solutions (pH 1-6.9) buffers or basic solutions (pH 7.1-14) indicated as an example. If desired the aqueous fluid may also contain one or more ionic or non-ionic surfactants, such as Triton X-100 ™, Tween^TM, Brij, Saponin and sodium dodecyl sulfate. Typically, the dewaxing fluid is heated. For example, if the incorporation medium is paraffin, which has a melting point between 50 and 57 degrees C, the fluid must be heated to a temperature higher than the melting point of paraffin, for example, 60 - 70 degrees C Typically, the fluid is heated in the fluid feed.

Referring also to Figures 5 and 6, the dewaxing station 140 also includes a fluid aspiration probe 150 arranged for pivotal movement of its distal end 152 to a central location 104 of a tray 80, when the latter is arranged in the dewaxing station 140. Probe 150 comprises a hollow tube connected via conduit (not shown) and suction pump 157 to a dewaxing agent separator (not shown) in which the dewaxing fluid is returned to fluid supply 32 where it is heated by A heater, as needed, is filtered in a filter 154 to remove cells that may have been dislocated during the dewaxing process, and reused. If desired, the accumulated paraffin can be removed, for example, by surface cleaning ("skimming"). The probe 150 must have sufficient freedom of movement between a deployed position where the probe is located adjacent to the central location 104 of the tray, and a position parked on top of the slide plate tray so as not to interfere with the movement of the slide. tray and slide plates in and out of dewaxing station 140.

A feature and advantage of the present invention, particularly compared to conventional bath-type dewaxing stations, is that potential cross-contamination between slides, for example, by the possibility of carryover of cells from one plate to another, is eliminated since the sample plate holders are subjected to freshly filtered dewaxing fluid only, and the horizontally spaced, coplanar orientation of the slide plates in the tray, prevents possible cross-contamination by cell carryover

ES 2 425 689 T3 between plates during the paraffinization process. Furthermore, the dewaxing process is made more efficient by the use of the hot dewaxing agent.

Referring in particular to Figure 7, coloring station 160 comprises a modular compartment and includes two or more wash and colorant dispensing nozzles 162. In a preferred embodiment of the invention, the coloring station comprises a pair of colorant dispensing nozzles 162, which are driven in a stepwise fashion along an axis 163 by a drive spindle 165 and a linear motor (not shown). from a pair of slide plates or another slide. The colorant dispensing nozzles 162 are selectively connected via valves and conduits and positive pressure pumps (not shown) to colorant reservoirs 168A, B, C, and alternatively the nozzles are selectively connected to a source of rinse liquid, typically DI optionally including a surfactant.

A fluid aspiration nozzle 170, similar to fluid aspiration nozzle 150, is disposed in coloring station 160 and is pivotally movable between a work station in which the distal end 172 of the nozzle is adjacent to the central location 104 of a tray in a staining station 160, and a parking position above the tray and the slide plates, in order not to interfere with the movement of a tray and slide in and out of the staining station 160. The suction nozzle 170 is connected via tubing (not shown) and a suction pump (not shown) to a waste container 38. As with the dewaxing station, the horizontal, coplanar orientation of the slide plates in the tray prevents cross-contamination of the plates during the staining process.

Coating station 180, which also comprises a modular unit, may comprise a fluid coating dispenser for applying a conventional fluid coating, such as that described in US Patent No. 2004/0092024 A1, filed April 26, 2002, entitled "Automated Coverslipper".

Alternatively, and in a preferred embodiment, as shown in Figures 8 and 8A, the coating station 180 comprises a cartridge or magazine 218 having an open dispensing end 220 '. Warehouse 218 defines a substantially rectangular box 222 ', in which cover glass plates 210 are stacked in a substantially vertical arrangement. A transfer mechanism, generally indicated 224, removes the top, or top cover glass plate 210 from the case 222 ', and places it on the glass slide 56 in the standby position. In a preferred embodiment, the transfer mechanism 224 includes a suction cup 226 suspended from a guide 228 and alternately driven along the guide by a linear motor and drive device 229. A vertically movable movable stem 230 extends through the bottom of the box 222 ', pushes the stack of glass plate liners into contact with the suction cup 226, so that the suction cup 226 engages with the plate of glass. cover glass 210. The pusher 230 is then retracted so that the stack of cover glass plates 210 is separated from the upper cover glass plate which is retained by the suction cup 226. The suction cup 226 is forced to advance, then along the guide 228 above the selected slide plate, and the suction cup is forced to release the cover glass plate onto the slide plate. The suction bowl is then returned to the top of the magazine 218, and the pusher 230 is again activated to push the stack of cover glass plates 210 into contact with the suction bowl 226, and the process is repeated.

The cover glass plates 210 each have a substantially flat top and bottom surface and a substantially rectangular configuration, the length and thickness being slightly less than the sample slide 56.

In an especially preferred embodiment, each of the cover glass plates 210 is coated, in turn on its lower surface, with a dry-activatable adhesive. In this case, a fluid dispensing nozzle 232 is supported by actuator 228 in advance of the cover glass plate 210 to apply an adhesive activating fluid to the surface of the sample slide. Preferred adhesives include Permount^TM (Fisher Scientific, Pittsburgh, PA) or ShurMount^TM (Triangle Biomedical, Durham, NC), which can be activated by a low viscosity fluid, such as toluene or xylene. An advantage of using adhesive coated glass cover plates and low viscosity activating fluid, such as xylene, is that air pockets, i.e. between the slide plates 56 and the cover glass plates 210, are avoided. US Patent No. 6,759,011 B1, filed July 6, 2004, further describes pre-glued liner elements.

The slide plate tray 54 is transported between said workstations by means of an XYZ transport and a lifting mechanism. With particular reference to Figures 1A, 1B, 9, 10A and 10B, the transport lift mechanism comprises a support table for the storage tray 60 comprising a generally rectangular frame 62 mounted slidably on the lift rail. 64. The frame 62 is connected by means of a bracket 74 on the elevator drive assembly 76 driven by one of an elevator transmission and with a drive motor 78. A counterweight 5 is arranged to compensate for the weight.

ES 2 425 689 T3 from the slide tray and smooth the acceleration and deceleration forces on the slide tray.

Referring in particular to Figures 9, 10A and 10B, the slide plate support table 60 also includes an XY loading / unloading mechanism 110 that includes a stepper motor controlled by drive systems 112A, 112B carrying supports. upwardly directed supports 114A, 114B to contact downwardly directed brackets 116A, 116B on a tray 80, to move tray 80 in and out of the lift and transport mechanism and in and out of a selected work station, as will be described in detail below.

In order to ensure that each tray is properly positioned at a work station, the transport mechanism for the elevator includes proximity sensors such as optical sensors 118 or microswitch sensors (not shown). Hall effect type sensors can also be used.

The operation of the apparatus indicated above will be explained below.

Referring to FIG. 11, specimen carrier plates 56 are placed on plate carrier trays 54. Plate carrier tray 54 is loaded into garage 60 through port 66. The transport and lifting mechanism is indexed just below plate tray 54 and the plate tray forced into the garage at a position where the downwardly extending bracket 116A is supported on tray 54 it is displaced past the upwardly extending bracket 114A supported on the lift / transport mechanism. The elevator / transport mechanism is then indexed in the vertical direction to align the elevator floor approximately level with the bottom of tray 54, and the tray is placed on the elevator / transport by retracting bracket 114A. The lift / transport mechanism is then moved vertically in the "Z" direction to a position adjacent to the technical processing station 42. The slide plate tray is then pushed by the holder 114B in an "X" direction into the heat treatment station 42, where the tray has been deposited. The holder 114B is removed, the tray carrying the slide plates with the samples is subjected to heat treatment for a predetermined period of time at a predetermined temperature, that is, to soften the paraffin on the slide plates. The elevator / conveyor mechanism is then adjusted substantially upward to align the elevator floor at approximately the level of the bottom of tray 54 at heat treatment station 42, and bracket 114B is retracted to drag the tray carrying the heat treated slide plates out of the treatment station 42, and then the tray is transported, as before, by the conveyor / elevator mechanism to the waxing or dewaxing station 44, where the tray is deposited at the station 44, and the slide plates are sprayed with heating water or a dewaxing fluid to remove the paraffin. Typically, alternating groups of slide plates are flooded with dewaxing fluid from nozzles 142 in a timed sequence. The dewaxing fluid is collected at the bottom of tray 80, where it is withdrawn by suction nozzle 150, and filtered through a 1 micron filter and recycled. To prevent excessive foaming of the aspirated dewaxing agent, the waste container 34 is preferably arranged in communication with the atmosphere.

The suction nozzle 150 is retracted, the slide plate tray 54 supporting the dewaxed specimen slide plates is removed from the dewax station 140, and transported, as in the previous case, by the transport / elevator device to staining station 160, where a selected stain is applied to individual slide plates. Selected stains include hematoxylin, eosin, and any other chemical stain usable to enhance the morphology of the tissue sample. Excess coloring and washing or draining is removed from the bottom of the tray by means of a suction nozzle, which is lowered into the center of the tray, and directed to the waste. Thus, fresh dye is always used, so that the problems that previously appeared in conventional bath-type staining devices, including cross-contamination of the slide plates, oxidation of stains and / or depletion of staining activity, are removed.

The suction nozzle is retracted and the colored plates are removed from the coloring station 160, and the tray can be transported back to the drying / heat treating station 120 for drying for a controlled period of time at a controlled temperature. Thereafter, the colored plates are removed from the treatment station 120, and transported as before, by the conveyor / elevator system to the coating station 180, where a glass coating is attached to the top surface of the plates. The lift / transport system then moves the coated plates to a storage position in garage 60, or the tray can be returned to the portal position where the tray is removed.

Referring to Figure 12 of the drawings, the apparatus 220 of the second embodiment of the invention, like the apparatus of the first embodiment, functions as a component or module of a system 222. The system 222 also includes fluid containers 232 , 234, 236 and corresponding equipment.

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As in the case of the first embodiment, the apparatus 220 comprises a frame that supports a stack of workstations comprising, for example, one or more drying or heat treatment stations or modules, a wax removal station or module. or dewaxing, one or more coloring stations or modules 260 and a coating station or module 280 arranged in a tower 250. However, in the second embodiment, the heat treating station and the dewaxing station are combined into a single module 220. A lift and transport mechanism 252, similar to the lift and transport mechanisms 52 discussed above, is disposed adjacent to tower 250 to transport a slide tray 54 (see Figure 13) designed to transport a series of individual specimen slide plates 56 from a tray storage station, through drying / heat treatment, waxing, coloring and coating operations.

In the embodiment shown, the combined drying / heat treating and dewaxing station or module 240, station or module 260, and coating station or module 280 are arranged vertically on tower 250 and are controlled by a computer.

Referring to FIG. 13, the combined drying / heat treating and waxing / dewaxing station or module 220 comprises a technically isolated compartment into which a controlled amount of heat is supplied for drying the sample plates. Station or module 220 preferably comprises a modular unit and includes a pair of radiant heating panels 300, 302, arranged to direct radiant heat toward the surfaces of the sample plates. Heating the plates serves to dry the plates, soften the paraffin on the plates, and heat the dewaxing fluid applied to the slide plates, as will be described in detail below. Station or module 220 also includes one or a series of dewaxing fluid dispensing nozzles 242 directed downwardly at an angle relative to the sample carrier plates. Preferably, the dewaxing station 240 comprises two arrays of ten nozzles 242, each fed through a common manifold 246 with a suitable dewaxing fluid from a supply of dewaxing fluid 232 (FIG. 12). Alternatively, a pair of nozzles can be mounted on a movable device and advanced from one pair of plates to another pair.

Various dewaxing agents can be used and preferably comprise concentrated solutions of aqueous fluids, such as Collaterge.^TM (Colonial Chemical, S. Pittsburg, TN). Collaterge can be used as an effective dewaxing agent in a wide range of concentrations, but preferably it is used in a concentration of approximately 3-30 volume percent. If desired, the concentrated aqueous solution can also contain one or more ionic surfactants or non-ionic, such as Triton X-100^TM, Tween^TM, Brij, Saponin and sodium dodecyl sulfate. To facilitate removal of the incorporation medium, i.e., the wax, the slide plates and the dewaxing fluid must be heated. For example, if the incorporation medium is paraffin, which has a melting point between 50-57 degrees C, the slide plates should be heat treated or preheated to a temperature of approximately 85 degrees C. A feature and advantage of the second embodiment of the invention is that preheating the slide plates to sufficient temperature eliminates the need to separately preheat the dewaxing fluid. Preheating the slide plates, that is, softening the paraffin, improves the efficiency of the dewaxing step. Depending on the conditions and environment of the amount and type of wax, it may be sufficient to squeeze the dewaxing fluid onto the preheated slide plates, let the fluid work for a few seconds or minutes, and then wash off the fluid and the Wax the plates using, for example, deionized water from the water nozzles 248. If necessary, the slide plates covered with a dewaxing fluid can be heat treated, eg, for several minutes, eg about 5 minutes, before being washed. In this way, the paraffinization process is reinforced. Furthermore, less dewaxing fluid is required and it is not necessary to filter and recycle said dewaxing fluid. Instead, the dewaxed, depleted fluid can be passed directly to the drain, or it can be filtered and then passed to the drain.

Station 240 also includes a fluid aspiration probe 250 similar to 150 of the first embodiment, and pivotally disposed at its distal end to a central location of the tray when the latter is disposed at station 240. The probe 250 comprises a hollow tube connected via tubing (not shown) and a suction pump 257, so that the spent dewaxing fluid can be filtered on a filter (not shown) to remove cells that may have been dislocated. during the dewaxing process, and the fluid can be sent to waste. The probe 250 must have sufficient freedom of movement between the deployed position, in which the probe is located adjacent to the central location of the tray, and a parking station above the tray and the slide plate so as not to interfere with the movement of tray and plates in and out of station 240.

A feature and advantage of the above-described second embodiment of the present invention, particularly compared to conventional bath-type dewaxing stations, is that potential cross-contamination between slide plates, for example, by the possibility of the one-plate cell carrier to another, it is eliminated since the sample plates are subjected only to fresh deparaffinizing fluid, and the horizontal, coplanar orientation, The spaced spacing of the specimen slides on the tray prevents possible cross-contamination by dragging cells between plates during the paraffinization process. In addition, the

ES 2 425 689 T3 dewaxing process is made more efficient by preheating the plates and / or heating the dewaxing agent on the plates.

As in the case of the first embodiment, the second embodiment includes a coloring station 260, which is similar in construction and operation to the previously described coloring station 160, and a coating station 280 is similar to the coating station 180. previously described. The coloring system according to a second embodiment also includes an XYZ elevator and transport device similar to the previously described XYZ elevator and transport mechanism. Of course, in the case of the second embodiment, the apparatus may have one less station or module, and therefore, the timing and sequencing of movements between the various modules will be different from that described.

The operation of the above-described apparatus according to the second embodiment will be provided below.

Referring to FIG. 14, the specimen-bearing slide plates are placed on the plate tray, and the plate tray is loaded into the transport and elevator mechanism 252. The transport and elevator mechanism is then vertically displaced in a "Z" direction to a position adjacent to the heat treating and dewaxing station 242, at which station the tray is deposited. The tray carrying the sample carrier plates is subjected to heat treatment for a predetermined period of time at a predetermined temperature, that is, to soften the paraffin on the plates, remove water from the samples and adhere the tissues to the plate. . Once sufficiently heated, for example, to a plate surface temperature of 85 degrees C, the plates are covered with concentrated dewaxing agent, and heated for 5 minutes. If desired, alternating arrays of plates can be sprayed with fresh concentrated dewaxing agent from nozzles 242 in a timed sequence. The slide plates are then washed with deionized water to remove dewaxing agent, wash water, dewaxing agent and paraffin, which collect at the bottom of the tray can be removed by suction nozzle 250, filtered to remove solids of the resulting filtrate is passed to waste.

The aspiration nozzle 250 is retracted, the tray 54 carrying the dewaxed specimen slide plates is then removed from the heat treatment and dewaxing station 240, and is transported as before, by the conveyor / elevator device to the station. coloring tool 260, in which a selected color is applied to individual plates, as described above.

The colored plates are removed from station 260, and the tray can be transported back to the heat treatment station or module 220 for drying for a controlled period of time at a controlled temperature. After that, the colored plates are removed from the heat treatment station or module 220, and are transported, as before, by the conveyor / elevator system to the coating station 280, where a glass coating is attached. to the upper surface of the plates. Coated plates can be sent to the heating / drying station to accelerate curing. The elevator / transport system then moves the coated plates into storage or the tray can be removed from the system.

The software for operating the system is designated as “Run Time Executive”. One of the responsibilities of the Run Time Executive (“RTE”) application is to sequence and schedule the operations carried out by the different functional workstations on each microscope slide tray. The system can handle 25 of these trays at a time, each tray requiring operations carried out by one or more workstations and perhaps multiple visits to the same workstation. Trays are moved within an instrument by a single elevator and shuttle table. Together, this elevator in combination with the table can move a tray in the X and Z directions as needed. The instrument also contains a "parking garage" in which trays can be positioned while they wait for a workstation to become available or when all operations on them have been completed. The maximum number of trays, 25, is adapted to the number of parking slots in the garage.

The basis for all actions carried out on a tray is a user-selected protocol that, among other things, designates the required workstation operations and tray priority as "STAT" or normal. Using this protocol, the RTE prepares an ordered sequence of workstations to visit. Since there is only one elevator per table, it can be considered as a single server with multiple tasks to perform. When the program for this problem can be calculated, it is necessary to know that the arrival of trays to the instrument cannot be predicted. Similarly, users can change the priority of a tray at any time. With these factors in mind, the schedule is dynamically determined each time the lift / table becomes available for work. The lift / table's “job” is to move a tray from point A to point B. Thus, after completion of the movement, the lift / table is available. At this point, the RTE examines each system tray and creates a list of possible moves. The process is the following:

ES 2 425 689 T3

1. First, it will be determined whether a tray can be moved. Moving a tray can be done from either a workstation, set aside and ready for the next workstation, set aside and listed for removal, or listed to be set aside due to an abnormal situation.

2. If the tray can be moved, its next destination must be identified with planned sequence and must be checked for availability. A workstation is considered available if it is simultaneously empty and ready for operation. If more than one of the card workstations are available, the workstation that has been waiting the longest is the one chosen. If the tray's target workstation is not available, it will be directed to the parking garage. In these cases, the RTE is always resumed by the empty tray closest to the next target station on the tray.

Once a list of all possible movements has been prepared, the RTE selects the movement to be carried out. This selection is based on tray priority and in the event of a tie, the time of arrival (TOA) from the tray to the system (ie time of entry is the portal) is determinable. The rules that control the priority of a tray are as follows:

1. A tray is assigned the highest priority if it is currently in the barcode reading / plate detection station. The highest priority is assigned because the reciprocating table is involved with this station operation and until you have completed and moved the tray to its next station, no other movement can be assigned to the elevator / table.

2. The second highest priority is assigned to a tray with a user designated STAT priority.

3. The third highest priority is assigned to a tray that is either in the portal waiting to enter the system, or is in the garage waiting to be removed from the system. The priority is adapted to the cases in which the user is waiting for the instrument.

Four. The lowest priority is assigned to any tray that meets the other three criteria.

The software mechanics of this selection consists of a register in a dynamic assembly structure performed for each tray that can be moved. This record contains the tray identification, their assigned priority, and their tOa. The set is selected by priority and then TOA and the entry at the top of the list is the tray provided to the elevator / table for its work.

It is appreciated, therefore, that the present invention will provide an integrated system capable of high throughput of staining of biological samples on slide plates. Among the advantages of the present invention is the elimination of inversion and emptying baths, dewaxing and / or coloration, which tend to be displeased by oxidation and / or contamination by dislocated biological cells during the dewaxing process. Rather, the present invention uses clean, fresh, or constantly filtered dewaxing agent, or coloring agent, thus eliminating the possibility of cell carryover from one plate to another.

Additionally the reagent utilization is approximately the same based on the utilization per plate (350 µl) compared to the inversion and bath device, which is a surprising fact. Furthermore, the present invention discloses for the first time a high-performance system, integrated complement, for staining of microscope slides from the heat treatment stage passing through the coating stage, the procedure of which is not carried out by any other system. commercially available today.

Different changes can be made to the above-described embodiments without departing from the spirit and scope of the invention. For example, the apparatus may include two or more coloring modules or stations, two or more heat treatment modules or stations, two or more dewaxing modules or station, and / or two or more combined heat treatment and dewax modules or stations. , which can further increase performance. A particular characteristic and advantage of the present invention is that additional modules or stations can be added vertically without increasing the plant or base of the system. Alternatively, two or more additional towers or stacks or workstations 50A, shown in Figure 1, can be served by a single elevator / transport system. Other reagents can be used on the instrument to perform other tests, including those used for in situ hybridization (typically DNA / RNA probes), or immunohistochemistry (typically antibodies). However, other changes may be made to the invention, without departing from the spirit and scope thereof, the scope of the invention being defined by the appended claims which will be interpreted in light of the foregoing description. An automated apparatus for slide plate processing comprises a series of workstations arranged vertically in a stack and a transport / lift device for transporting slide plates between said workstations.

An automated apparatus for slide plate processing further comprises a computer for controlling said stations and said transport / elevator device.

ES 2 425 689 T3

According to another aspect, in an automated apparatus for the slide plate process, at least one of said stations comprises a heat treatment or drying station.

In another aspect, in an automated apparatus for slide plate processing, said heat treating or drying station comprises a convection heater.

In another aspect, in an automated apparatus for slide plate processing, at least one of said workstations comprises a dewaxing or dewaxing station.

In another aspect, in an apparatus for automated slide plate processing, said dewaxing or dewaxing station comprises a series of fluid dispensing nozzles arranged to deliver a dewaxing or dewaxing fluid onto said slide plates.

In another aspect, in an automated apparatus for slide plate processing, said dewaxing or waxing fluid comprises an aqueous fluid.

In another aspect, in an automated apparatus for slide plate processing, at least one of said stations comprises a coloring station.

In another aspect, in an automated apparatus for slide plate processing, said coloring station comprises a series of nozzles for applying selected colorants or wash fluids to said slide plates.

In another aspect, an automated apparatus for slide plate processing further comprises a series of coloring supplies selectively connected to said nozzles.

In another aspect, in an automated apparatus for slide plate processing, said work station comprises a coating station.

In another aspect, in an automated apparatus for slide plate processing, said coating station is adapted to apply a glass coating to said slide.

In another aspect, an automated slide plate processing apparatus comprises a slide plate tray for supporting a plurality of plates as they are transported through said apparatus.

In another aspect, in an automated apparatus for slide plate processing, said conveyor / lift device comprises an XYZ conveyor mechanism.

In another aspect, an automated apparatus for slide plate processing comprises a filter and recirculator for filtering and recirculating the waxing or dewaxing fluid.

In another aspect, an automated apparatus for slide plate processing comprises a series of stacks of work stations operatively disposed adjacent to a common conveyor / elevator device.

In another aspect, an automated apparatus for slide plate processing further comprises a heater for heating said aqueous fluid.

A dewaxing or dewaxing station for processing specimen slide plates comprises a container containing an aqueous dewaxing or dewaxing fluid; a heater for heating said aqueous fluid; a sprayer for spraying said hot aqueous fluid onto said slide plates; a manifold to collect and return the aqueous fluid to the receptacle and a filter to filter the aqueous fluid for reuse.

According to another aspect, in a waxing or dewaxing station, said filter is located between said manifold and said container.

According to another aspect, a waxing or dewaxing station further comprises a tray for supporting a series of spaced-apart sample slides in a substantially horizontal orientation.

According to another aspect, a waxing or dewaxing station further comprises a series of spray nozzles for spraying said waxing or dewaxing fluid from said slide plates.

ES 2 425 689 T3

According to another aspect, in a waxing or dewaxing station, said tray has an inclined bottom wall and further comprises an aspirator for drawing in the consumed waxing or dewaxing fluid from an adjacent low point of said inclined bottom wall. .

According to another aspect, in a waxing or dewaxing station said tray comprises a shelf to support said series of slide plates, spaced apart, above the bottom wall of the tray.

According to another aspect, in a waxing or dewaxing station said shelf comprises a series of supports that limit the axial, lateral and vertical movement of the slide plates in the tray.

According to another aspect, in a waxing or dewaxing station said rack is adapted to support a series of slide plates in separate alignments with each other.

Automated apparatus for slide plate processing comprising:

a series of workstations arranged vertically in stacks; and a transport / elevator device for transporting said slide plates between said workstations, wherein one of said workstations comprises a combined heat treatment or drying and dewaxing or dewaxing station.

In another aspect, an automated apparatus for slide plate processing further comprising a computer for controlling said stations and said transport / elevator devices.

In another aspect, in an automated apparatus for slide plate processing said combined heat treating or drying and dewaxing or dewaxing station comprises at least one radiant heater.

In another aspect, in an automated apparatus for slide plate processing, said combined heat treatment or drying and dewaxing or dewaxing station comprises a series of fluid dispensing nozzles arranged to supply a dewaxing or waxing fluid to said preheated microscope slides.

In another aspect, in an automated apparatus for slide plate processing, said dewaxing or waxing fluid comprises an aqueous type fluid.

In another aspect, in an automated apparatus for slide plate processing said dewaxing or dewaxing fluid comprises from about 3 to about 30 volume percent Collaterge^TM.

In another aspect, an automated apparatus for slide plate processing further comprises a coloring station.

In another aspect, in an automated apparatus for slide plate processing said coloring station comprises a series of nozzles for applying selected colorants onto said slide plates.

In another aspect, an automated apparatus for slide plate processing comprises a series of coloring supplies selectively connected to said nozzles.

In another aspect, an automated apparatus for slide plate processing further comprises a coating station.

In another aspect, in an automated apparatus for slide plate processing said coating station is adapted to apply a glass coating to said slide plate.

In another aspect, an automated slide plate processing apparatus further comprises a slide plate tray for supporting a plurality of plates as they are transported through said apparatus.

In another aspect, in an automated apparatus for slide plate processing said elevator / transport device comprises an X, Y, Z transport mechanism.

In another aspect, an automated slide plate processing apparatus comprises a series of work station stacks operatively disposed adjacent to a common elevator / transport device.

A combined heat treating or drying and waxing or dewaxing station for the sample slide processing comprises a radiant heater for heating the slide plates and a

ES 2 425 689 T3 sprayer for spraying waxing or dewaxing fluid from a reservoir onto said heated slide plates.

In another aspect, a combined heat treating or drying and dewaxing or dewaxing station further comprises a sprayer for spraying a washing fluid onto said slide plates.

In another aspect, a combined heat treating or drying and dewaxing or dewaxing station further comprises a tray for supporting a series of spaced apart sample slides in substantially horizontal orientation.

In another aspect, a combined heat treating or drying and dewaxing or dewaxing station further comprises a series of spray nozzles for spraying said dewaxing or dewaxing fluid onto said slide plates.

In another aspect, in a combined heat treating or drying and dewaxing or dewaxing station, said tray has a sloping bottom wall and further comprises an aspirator for aspirating spent dewaxing or dewaxing fluid from an adjacent low point of said sloping back wall.

In another aspect, in a combined heat treating or drying and dewaxing or dewaxing station said tray comprises a shelf for supporting said series of spaced apart slide poles above the bottom wall of slide plates.

In another aspect, in a combined station for heat treatment or drying and dewaxing or dewaxing said shelf comprises a series of supports that limit the axial, lateral and vertical movement of the slide plates in the tray.

In another aspect, in a combined heat treating or drying and dewaxing or dewaxing station said rack is adapted to support a series of slide plates in spaced apart alignments.

A procedure for the automatic preparation of tissue samples on microscope slides for pathological analysis comprises:

heat treating the tissue sample on the plate by causing the instrument to apply sufficient heat to the tissue for adherence to the slide plate;

dewaxing the tissue sample by contacting it with a dewaxing fluid at a temperature higher than the melting point of the paraffin and then washing the liquefied paraffin by dragging it; coloring the tissue sample by contacting it with a coloring reagent; and coating the slide plate.

In another aspect, in one method, said slide plate is provided with contact coating of the colored tissue sample on the slide plate with a pre-glued coating and an adhesive activating fluid.

In another aspect, in one process, said dewaxing fluid comprises water and a surfactant.

In another aspect, in one method, said coloring reagent comprises hematoxylin and eosin.