WO2021069121A1

WO2021069121A1 - Device and method for wetting biological material

Info

Publication number: WO2021069121A1
Application number: PCT/EP2020/069878
Authority: WO
Inventors: Xaver Einsle
Original assignee: Prime23 GmbH
Priority date: 2019-10-10
Filing date: 2020-07-14
Publication date: 2021-04-15
Also published as: DE102019215577B3

Abstract

A device for wetting biological material with at least one liquid, comprising a cartridge (1) with a cartridge bottom (2) and a cartridge top (3), wherein the cartridge (1) has at least two receiving regions (6), each of them for a respective holding device (7), wherein the holding device (7) serves to receive a slide (8) which carries the biological material, wherein the biological material arranged on the slide (8) can be wetted with the liquid inside the closed cartridge (1), and wherein the slide (8) inside the closed cartridge (1) can be at least partially lifted from the holding device (7) so that liquid located between the slide (8) and the holding device (7) can run off. Furthermore, a method for wetting biological material with at least one liquid is proposed.

Description

DEVICE AND METHOD FOR WETTING BIOLOGICAL MATERIAL

The invention relates to a device for wetting biological material with at least one liquid.

The invention also relates to a method for wetting biological material with at least one liquid.

Devices of the type in question are used, for example, for the automated processing of biological materials which are attached to a large number of microscopic slides and which are individually processed in incubation chambers. The main area of application is the diagnostic field of anatomical pathology.

A corresponding device is known, for example, from DE 102016225 885 A1. This describes a special holding device for a specimen slide, in which the specimen slide is tilted several times during processing in accordance with a protocol developed for this purpose, at fixed times and work steps in a continuous movement around the three coordinate axes. Thus, liquid that is located between the specimen slide and the holding device can drain off.

The problem with this device, however, is that the user has to open and close individual lids via a hinge on the multiblock in order to insert the specimen slide into the device. For example, in the case of an instrument with 36 individual holding devices, the user has to insert a slide 36 times individually. The advantage of this system is that an individual holding device and thus the respective specimen slide always lies correctly on the temperature control unit, which ensures good temperature transfer. In addition, the suction nozzle fastened in the lid is connected to the vacuum or suction unit and the gap between the holding device and the underside of the specimen slide, in which there is When raised, collect the reagents to be aspirated at the lower end of the slide. The disadvantage of this system, however, is that the user still has to do a lot of manual work by inserting and, after finishing the processing of the specimen slide, just as much manual work when removing, for example, 36 individual specimen slides. As a result, this device is relatively time consuming to operate.

A device for wetting biological material is also previously known from US Pat. No. 8337786 B2. This also has the disadvantages described above or similar.

The present invention is therefore based on the object of designing and developing a device for wetting biological material with at least one liquid of the type mentioned at the beginning in such a way that handling is facilitated with structurally simple means. Furthermore, a method for wetting biological material with at least one liquid is to be specified.

According to the invention, the above object is achieved by the features of claim 1. According to this, the device in question for wetting biological material with at least one liquid, a cassette with a cassette bottom and a cassette lid, wherein the cassette has at least two receiving areas for each holding device, the holding device serving to receive a slide that holds the biological Carries material, wherein the biological material arranged on the slide is wettable with the liquid within the closed cassette and wherein the slide within the closed cassette can be at least partially lifted off the holding device in order to allow liquid that is between the slide and the holding device to drain off located to enable.

A particular advantage of this embodiment is that the cassette is portable, ie can be handled as such by the user. The user can place the specimen slide on the respective holding devices and pass the cassette through Close the merging of the cassette lid and the cassette base. The closed cassette can then be operatively connected to a console or to another device, for example placed on it or inserted into it.

Advantageously, pressure points, in particular pressure points made of a flexible material, preferably made of a plastic, in particular silicone, can be arranged in or on the cassette cover in such a way that they each press the holding devices in the direction of the cassette bottom. The pressure points can be implemented as point-like elevations on the cassette lid, which are each in contact with a region of the holding devices when the cassette is closed by connecting the box bottom and the cassette lid to one another. This has the advantage that the holding devices can be pressed onto the temperature control devices of a console via the pressure points, so that the various required temperature degrees can be transferred directly to the holding devices, regardless of an exact, linearly shaped cassette base. Alternatively or additionally, it is conceivable that the holding devices are movably arranged within the cassette.

In the manner according to the invention, it has been recognized that the underlying object can be achieved in an amazingly simple manner by arranging several holding devices together in a cassette. In the simplest case, the cassette can have two holding areas. However, it is conceivable that the cassette can accommodate, for example, four, eight or even considerably more holding devices, so that a large number of specimen slides can be handled at the same time. The “multiple system” according to the invention simplifies the entire process for the user, since the cassette can first be equipped with the specimen slides and then operatively connected to a console or to another device.

The holding devices can advantageously be removed from the cassette. This simplifies the handling even more, since the Slides can initially be placed on a holding device and can then be placed in the cassette together with them.

In a particularly advantageous manner, at least a part of the receiving area can be formed by a passage in the cassette base. In other words, the cassette bottom can be designed like a grid, the openings serving as receiving areas for the holding devices.

According to a further advantageous embodiment, a console can be arranged, the cassette base being able to be placed on the console and removed from it. It is conceivable that the console has a deflection device so that each specimen slide can be lifted, preferably individually, within the closed cassette. Alternatively or additionally, a temperature control device can be arranged on the cassette and / or on the console for each receiving area. Thus, the console could be used for temperature control, i.e. for heating or cooling, as well as for lifting the slide.

In a particularly advantageous manner, the cassette lid and the cassette bottom can be connected to one another via magnetic forces, in particular via at least one permanent magnet. Such a connection has the advantage that the cassette lid and the cassette base are precisely aligned with one another when they are brought together by the magnetic forces, so that, for example, tilting is prevented. Accordingly, such a construction is safe and easy to handle even with laboratory gloves. Alternatively or additionally, the cassette lid and the cassette base can be connected to one another in a form-fitting and / or force-fitting manner, in particular via a clamping and / or latching mechanism, for example via a bayonet lock. The advantage of a corresponding configuration is that it is simple and therefore inexpensive to manufacture. Furthermore, it is ensured that the cassette lid and the cassette base are extremely securely and firmly connected to one another. To further simplify handling, it is conceivable that the cassette and the console can be connected to one another via magnetic forces, in particular via at least one permanent magnet. As already stated above, this has the advantage that the elements to be connected are aligned with one another, which considerably simplifies handling. In a further advantageous manner, the cassette and the console can be connected to one another in a form-fitting and / or force-fitting manner, in particular via a clamping and / or latching mechanism, for example via a bayonet lock. A secure connection between the cassette and the console can thus be realized with structurally simple means. Both in the case of a connection via magnetic forces and in the case of a form-fitting and / or force-fitting connection, the holding devices can be pressed against any temperature control devices in such a way that there is a uniform thermal coupling.

In order to be able to remove the liquid or the reagents which are used for treating the samples located on the slide, a suction device can be arranged. This can comprise suction means arranged on the cassette lid and / or on the cassette bottom. It is conceivable that the suction means are connected to a vacuum device, for example a pump, via a preferably common fluid connection. Specifically, the fluid connection could be designed, at least in some areas, as a hose connection or a push-in pipe connection. In a particularly advantageous manner, push-off means could be provided in order to interrupt the fluid connection of a suction means with the vacuum device. The push-off means could be formed by a pretensioned push-off element. The push-off means can be designed in such a way that the pressure exerted by it is only slightly above the pressure required for pushing off or squeezing off, so that the push-off means can be released with as little force as possible. For this purpose, the push-off means could be movable against its bias by a release device, for example an electromagnet or an electric motor, in order to release the fluid connection. According to a further advantageous embodiment, indentations, on which the side wings rest, can be formed on the holding device and on the receiving area. With this construction it can be achieved in a simple manner that the holding devices are movably arranged in the cassette. In a particularly advantageous manner, the depth of the indentations could be greater than the thickness of the side wings. For example, the side wings could have a thickness of 0.2 mm to 1.0 mm, in particular a thickness of 0.4 mm to 0.8 mm, preferably a thickness of 0.6 mm, and the depth of the indentations in the range of 1.0 mm. 1 to 1.9 mm, in particular from 1.3 mm to 1.7 mm, preferably 1.5 mm. In such a configuration, pressure points could be formed on the cassette cover which press the holding devices on the side wings in the direction of the cassette bottom when the cassette is closed. This achieves the advantage that the holding devices can be pressed onto a temperature control device when the cassette is connected to the console, even if the cassette bottom is not flat on its side facing the console, for example it is convex and / or concave due to the manufacturing process.

With regard to the method, the underlying object is achieved by claim 21. According to this, the method for wetting biological material with at least one liquid, using a device according to one of claims 1 to 20, has the following method steps:

- Insertion of slides carrying the biological material on a respective holding device before or after the holding devices have been inserted into the receiving areas of the cassette bottom,

- Merging of the cassette lid and the cassette base to close the cassette,

- Connect the cassette to the console, and

- Introducing the liquid and / or the liquids into the holding devices.

According to the invention, it has been recognized that the wetting of biological material can be considerably simplified by first introducing the slide carrying the biological material into a cassette which is then closed. The specimen slides can initially each be placed on their own holding device and these can then be placed in the holding areas of the cassette base. Alternatively, it is conceivable that the holding devices are first positioned in the holding areas and then the specimen slides are applied. Regardless of this sequence, the closed cassette equipped with the specimen slides can be connected to the console, for example inserted into it or placed on it. The liquid can then be introduced into the holding devices so that it comes into contact with the biological material.

Furthermore, additional steps can be carried out, for example heating and / or cooling the samples, moving the slides, suctioning off excess liquid, etc.

It is pointed out that the described device according to the invention can have a procedural characteristic. The features and advantages described with regard to the device can therefore expressly be part of the method according to the invention.

There are now various possibilities for designing and developing the teaching of the present invention in an advantageous manner. For this purpose, on the one hand, reference is made to the claims subordinate to claim 1 and, on the other hand, to the following explanation of preferred exemplary embodiments of the invention with reference to the drawing. In connection with the explanation of the preferred exemplary embodiments of the invention with reference to the drawing, generally preferred configurations and developments of the teaching are also explained. Show in the drawing

Fig. 1 in a schematic representation a perspective view of a

Embodiment of a device according to the invention comprising a console, 2 in a schematic representation a perspective view of a further exemplary embodiment of a device according to the invention comprising a console,

3 in a schematic representation a perspective view of a further exemplary embodiment of a device according to the invention comprising a console,

Fig. 4 in a schematic representation, a perspective view of a

Cassette of a device according to the invention,

Fig. 5 in a schematic representation, a perspective view of a

Folding device of a device according to the invention,

6 in a schematic representation a perspective view of a further folding device of a device according to the invention,

7 in a schematic representation a perspective view of a further folding device of a device according to the invention,

8 in a schematic representation a perspective view of a further folding device of a device according to the invention,

9 shows a schematic representation of a perspective view of a

Cassette bottom of a device according to the invention,

10 shows, in a schematic representation, a perspective view of a further cassette bottom of a device according to the invention,

11 is a schematic representation of a perspective view of a further cassette bottom of a device according to the invention,

12 is a schematic representation of a perspective view of a cassette cover of a device according to the invention, 13 shows, in a schematic representation, a perspective view of part of a suction device of a device according to the invention,

14 in a schematic representation a perspective view of part of a suction device of a device according to the invention,

15 shows, in a schematic representation, a perspective view of part of a suction device of a device according to the invention,

16 shows, in a schematic representation, a perspective view of a suction means of a device according to the invention,

FIG. 17 shows, in a schematic representation, a further perspective view of the suction means according to FIG. 16,

18 shows, in a schematic representation, a further perspective view of the suction means according to FIG. 16,

19 shows, in a schematic representation, a further perspective view of the suction means according to FIG. 16 with an object carrier arranged for it,

20 in a schematic representation a perspective top view of part of a console of a device according to the invention

FIG. 21 shows, in a further schematic illustration, a perspective view from below of the part of a console according to FIG. 20,

22 in each case in a schematic representation a perspective view of a deflection device and a deflection device with an attached slide of a device according to the invention, 23 shows, in a schematic representation, a perspective view of the deflection device of a device according to the invention,

24 shows, in a schematic representation, a perspective view of a cassette cover for use with the deflection device according to FIGS. 22 and 23,

25 shows, in a schematic representation, a perspective view of a receptacle for a temperature control device of a device according to the invention,

26 shows, in a schematic representation, a perspective view of a further cassette of a device according to the invention,

27 in a schematic representation, a perspective view of part of a suction device of a device according to the invention,

FIG. 28 shows, in a schematic representation, a perspective view of the part of a suction device according to FIG. 27,

29 shows, in a schematic representation, a perspective view of a suction means of a device according to the invention,

FIG. 30 shows, in a schematic representation, a further perspective view of the suction means according to FIG. 29,

31, in a schematic representation, a further perspective view of the suction means according to FIG. 29, and

32 shows a block diagram to illustrate an exemplary embodiment of the method according to the invention. In the figures, the same elements are provided with the same reference symbols. Furthermore, in order to improve clarity, not all elements are provided with a reference number in every figure.

FIGS. 1 and 2 each show, in a schematic representation, a perspective view of an exemplary embodiment of a device according to the invention. This has a cassette 1 consisting of a cassette bottom 2 and a cassette cover 3. In Fig. 1, only the preferably transparent cover disk 4 of the cassette cover 3 is shown to simplify the representation.

The cassette 1 is arranged on the console 5 and has a total of four receiving areas 6 for one folding device 7 each. A slide 8 is placed on each of the folding devices 7.

In a first embodiment according to FIG. 1, the console 5 has a first plate 9 and a second plate 10, which are designed as individual components. In a second embodiment according to FIG. 2, the first plate 9 and the second plate 10 are designed in one piece, for example fused to one another. For the sake of simplicity, the first plate 9 and the second plate 10 are further described below, even if this can be a single plate.

It can be clearly seen here that the first plate 9 is formed at an angle. The slope can be in the range from 1.5 ° to 5 °, in particular from 2 ° to 4 °, preferably 3 °. The inclined course of the first plate 9 can be designed independently of the further construction of the device. It is thereby achieved that liquid which is pipetted into the folding device 7 runs in the direction of the overflow channel 11 formed in the cassette base 2. Furthermore, the incline enables any bubbles in the liquid to rise better.

Furthermore, four electric motors 12 are formed which serve as a drive for a deflection device 13 via which the specimen carriers 8 in the cassette 1 can be deflected. In addition, the suction device 14 can be seen, which comprises the suction means 15 by means of which the liquid can be suctioned off from the individual holding devices 7. For this purpose, the suction means 15 are connected via a fluid connection 16, for example to a pump or another device for generating a negative pressure. The fluid connection 16 is designed as a hose connection in some areas. In order to interrupt the hose connection, push-off means 17 are provided, which here comprise electromagnets 18, any other control elements being conceivable. Control means 19, for example electronic boards, are also arranged.

In the exemplary embodiment shown here, the cassette 1 and the console 5 are connected to one another via magnets 20 (not shown). Furthermore, the cassette bottom 2 and the cassette lid 3 are connected to one another via magnets 21.

3 shows, in a schematic representation, a perspective view of a further exemplary embodiment of a device according to the invention. The device generally corresponds to the device shown in FIGS. 1 and 2, so that reference is made to the description in this regard. The main difference here is that the cassette base 2 and the cassette lid 3 are connected to one another in a form-fitting and force-fitting manner, namely via a first positioning hook 60 which acts in the manner of a bayonet lock. Furthermore, the cassette 2 is positively and non-positively connected to the console 5 via two second positioning hooks 61, which act as latching hooks.

Another difference is that the suction means 15 are each positioned in a corner of the holding devices 7.

4 shows, in a schematic representation, a perspective view of a cassette 1 of a device according to the invention. The cassette 1 has several receiving areas 6 in which suction means 15 designed as suction nozzles are arranged. The cassette base 2 has the functions of “picking up, positioning and holding” a defined number of holding devices 7. Furthermore, because of its magnets 20, which cannot be seen in FIG. 4, the cassette bottom is used for automatic positioning in relation to a console 5.

The cassette lid 3 has the functions of “receiving and holding a certain number of suction means 15 or suction nozzles” corresponding to the number of folding devices 7 in the cassette bottom 2. Furthermore, by means of the pressure points 34 of the cassette lid 3, the folding devices 7, possibly formed with side wings 22, which are loosely suspended or stored in the receiving areas 6, are pressed into the recessed positioning points of the cassette bottom 2 provided for this purpose. If necessary, the cassette lid 3 is used at the same time to press the bottom of the folding devices 7 onto the individual temperature control devices 23, provided that these are arranged. Furthermore, the cassette cover 3 can be covered with a cover disk 4, which can be transparent or partially transparent, in order to reduce the partial evaporation of the same resulting from the heating of the reagents.

The cassette 2 is closed after the specimen slides 8 to be processed have been inserted into the folding devices 7 outside the device. “Closing” means that the cassette lid 3 is placed on the cassette bottom 2 and then automatically positioned relative to one another and at the same time closed with magnetic force due to the magnetic tensile forces of the magnets 21 in both subcomponents. Alternatively, the specimen slides 8 can also be inserted from above into the already closed cassette 2 when the cover disk 4 of the cassette cover 3 has been removed. This is a realistic option and can be determined by the user. The immune staining process is not affected.

After the dyeing process has been carried out, the cassette 2 is removed from the console 5. Removal means that the forces of the magnets 20, which are fastened in the console 5 and in the cassette 1, for example in the longitudinal struts 24, must be overcome by manual or mechanical gripping. Then the cassette 1, consisting of the cassette lid 3 is opened manually from the cassette bottom 2, whereby the forces of the magnets 21 in the cassette lid 3 and the cassette bottom 2 are overcome in terms of force have to. The slides 8 can then be removed with or without holding devices. In the closed state and equipped with a cover disk 4, an object carrier 8 which is only loosely in the holding device 7 cannot fall out of the cassette 1 even if it overturns.

5 to 8 each show, in a schematic representation, a perspective view of different holding devices 7 of a device according to the invention.

The holding device 7 is one of the central components of the device according to the invention. For this reason, it is advantageous to have a holding device 7 which enables simple handling and at the same time meets the requirements demanded by the specialist staff who assess the immunostaining. These requirements include: as little background noise as possible, high specificity, repeatability, and clarity. The holding devices 7 described in FIGS. 5 to 8 not only make it possible to meet these requirements, but also enable the use of holding devices 7 of different sizes, so that an adaptability to the size of a slide 8 but also an adaptation to the size of the staining area is created on the slide 8. With a short holding device 7, for example only 30 mm long instead of the usual 55 mm to 60 mm, the consumption of reagents is automatically limited. Only about half of the slide 8 is wetted with reagents. The biological material to be stained must therefore be attached to the front, wettable surface of the slide 8. With regard to the consumption of reagents, it is important that the slide 8 does not touch the side walls 26 of a holding device 7 in order to avoid that capillary forces but also other forces of surface tension get reagents into the lateral gaps between the inside of a side wall 26 and the sides of the Slide 8 crawl.

For this reason, various holding devices 7 are shown in FIGS. 5 to 8, each of which contains different concepts of spacers 25. In the case of the holding device 7 shown in FIG. 5, the side walls 26 run obliquely and have elongated side wings 22 which are used for positioning. Furthermore, elongated or punctiform, embossed support shoulders 27 are formed on the rear wall 28 and a support point 29 on the tear-off edge 30 for the specimen slide 8.

The folding device 7 shown in FIG. 6 has inclined side walls 26, with, in contrast to the folding device according to FIG. 5, no side wings 22 being present. Furthermore, lateral spacers 25 are formed on the two inner sides of the folding device 7. In order to accommodate the specimen slide, support points 29 are also provided, which can be implemented as punctiform impressions in the floor. The suction of the liquid takes place ideally from one of the two corners on the rear wall 28.

The folding device 7 shown in FIG. 7 has inclined side walls 26, with, in contrast to the folding device according to FIG. 5, likewise no side wings 22 being present. Furthermore, only a single lateral and only a single rear-wall spacer 25 are arranged. In the interior of the folding device 7, in the corner provided for sucking off the liquid, support points 29 are formed on the base, in particular embossed into it.

The folding device 7 shown in FIG. 8 has at least essentially vertical side walls 26, ie the side walls 26 run essentially at an angle of 90 ° with respect to the bottom of the folding device 7. Furthermore, there are relatively short side wings 22 on the side walls 26 and bulges 31 intended. The bulges 31 are used for pipetting in or for introducing a pipetting needle. In this exemplary embodiment, the liquid is sucked off in the middle or from a corner.

FIGS. 9 to 11 show different configurations of a cassette bottom 2.

The concept of the cassette floor 2 integrates the functions:

- Positioning of the folding devices 7 in the cassette bottom, - Integration of the overflow channel 11, which is at the same time a spacer to the housing of the deflection device 13

- Housing and cavity for the deflection device 13 or for its tip per holding device 7,

- Admission of magnets 20, 21 with the three functions

1. Positioning of the cassette bottom 2 with the holding devices 7 on the console 5,

2. Close the cassette bottom 2 with the cassette lid 3, and

3. Positioning of the cassette 1 in the console 5. Each time the magnetic forces come into effect with corresponding magnets in the other subcomponents.

Individual placement of the holding devices 7, which are mounted in the cassette bottom 2 so that they can move easily, onto the temperature control device 23

The cassette bottom 2 shown in FIG. 9 is adapted to the holding device 7 shown in FIG. 5. Indentations 32 are provided in / on the longitudinal struts 24 for positioning in the xy plane of a holding device 7 by means of the elongated side wings 22. Advantageously, the depth of the indentations 32 can be greater than the thickness of the side wings 22, so that the holding devices are movably supported within the receiving areas 6. As a result, an underside of the cassette bottom 2 that is not perfectly flat can be compensated for, so that the holding devices 7 essentially completely rest on any temperature control device 23 of the console 5. For this purpose, the holding devices 7 can be pressed on their side wings 22 by pressure points 34 of the cassette lid 3 in the direction of the cassette bottom 2. Furthermore, the bevels 33 in the longitudinal struts 24 are adapted to the inclined side walls 26 of the holding device 7 according to FIG. 5. The indentations 32 also serve as pressure-absorbing surfaces that the cassette lid 3 exerts with its pressure point 34. Furthermore, the overflow channel 11 and a recess 35 are shown. A corresponding counterpart of the cassette cover 3 engages in the recess 35 so that the two parts are positioned or aligned with one another. It should be noted that the receiving areas 6 through Recesses are formed so that the bottom of the holding devices 7 is not covered by the cassette bottom.

The cassette bottom 2 shown in FIG. 10 is adapted both to the holding device 7 shown in FIG. 6 and to the holding device 7 shown in FIG. 7. The bevels 33 formed in the longitudinal struts 24 serve as a positioning aid and prevent the holding devices from slipping through the receiving areas 6. The positioning of a holding device 7 can be variable in the longitudinal plane. The suction can take place from one of the two corners of the holding device 7.

The cassette bottom 2 shown in FIG. 11 is adapted to the holding device 7 shown in FIG. 8. Indentations 32 are formed in / on the longitudinal struts 24 for positioning in the x-y plane of a holding device 7 by means of the side wings 22. The indentations 32 are adapted to the respective side wings 22 in such a way that they also serve as a pressure-absorbing surface of the cassette lid 3. Additional indentations 36 are also provided at the points of the longitudinal struts 24 at which indentations 31 for a pipetting needle are present in the holding device 7 on the left and right. Advantageously, the depth of the indentations 32 can be greater than the thickness of the side wings 22, so that the holding devices are movably supported within the receiving areas 6. As a result, an underside of the cassette bottom 2 that is not perfectly flat can be compensated for, so that the holding devices 7 essentially completely rest on any temperature control device 23 of the console 5. For this purpose, the holding devices 7 can be pressed on their side wings 22 by pressure points 34 of the cassette lid 3 in the direction of the cassette bottom 2.

12 shows, in a schematic representation, a perspective view of a cassette lid 3 of a device according to the invention, in particular the side of the cassette lid 3 facing the cassette bottom 2. The cassette lid 3 integrates the functions:

Exerting a pressure on the side wings 22 of the holding devices 7 by means of an in particular soft and / or flexible pressure point 34. This can preferably be made of silicone or rubber and / or be implemented by a spring, in particular a compression spring, and be arranged in / on the cover struts 37. The two side wings 22 are thus pressed into the indentation 32 of the cassette base 2 on the one hand and the base of each holding device 7 is pressed individually onto the temperature control devices 23 on the other hand.

- Picking up and holding the suction means 15 or suction nozzles

- Receiving an in particular transparent or partially transparent cover plate 4

- Receipt of a positioning screw or a positioning pin in order to be able to rotate a specimen slide 8 in a holding device 7 around its longitudinal axis (for suctioning off the liquid in a corner).

13 to 19 show the individual components of a suction device 14 of a device according to the invention.

In Fig. 13, the pressing means 17 and the suction means 15 are shown. The push-off means 17 serve to close off a deformable hose 16 or a deformable tube by being squeezed together from the vacuum suction. This makes it possible to replace a vacuum valve, which is more expensive and complicated to manufacture. However, the invention also allows the use of a vacuum valve.

The push-off means 14 shown here have a push-off element 14 designed as a lever 41, which is anchored at one end by means of a pin. The other end of the lever 41 is connected to the armature 42 of an electromagnet 18, for example. For example, on the underside of the lever 41, just in front of the armature 42, a compression spring 43, alternatively also a tension spring, is attached, for example, which constantly presses the lever 41 upwards (or alternatively in one direction). Above the lever 41, preferably at a 90 ° angle, lies a deformable hose section 16 or pipe section which is intended to be compressed by the lever 41. For this purpose, it is necessary, if the lever 41 has a pressing effect on the underside of the deformable hose 16 / tube, and a device causing a counterpressure to be attached to the opposite side of the hose 16 - in the case described above. This is a Cover 44 is provided for the housing 45 of the suction device 14, which has a projection 46 or a nose which presses from above onto the deformable hose 16 or the tube and slightly squeezes the upper side of the hose 16 downwards in the idle state. The lever 41, which presses from the bottom upwards, thus closes the hose 16 constantly in the idle state and is opened by activating the electromagnet 18, as a result of which the lever 41 is pulled downwards. Ideally, the compression spring 43 is slightly stronger in pressure than the resistance of the deformable hose 16 or the pipe, so that a relatively small or weak electromagnet 18 is sufficient to open the hose 16 and enable suction by means of vacuum.

An alternative or additional push-off means 17 is shown in FIGS. This has a motor 38, for example an electric motor 38, for raising or lowering a lever 41, a holder 39 for fastening the motor 38 to a plate 9, 10 and a static or movable abutment 40. The motor 38 can act on the lever 41 via a thread, so that the latter can be moved linearly by a rotary movement of the motor 38. A rotation caused by the motor 38, for example a left-hand rotation, allows the lever 41 to be extended and squeezes the flexible hose 16 against the abutment 40. Thus, in the completely squeezed state, the possibly constantly acting vacuum is blocked and unwanted suction is prevented. If the lever 41 is retracted by causing the motor 38 to rotate in the opposite direction, for example a clockwise rotation, the hose 16 opens into its original shape and the vacuum comes into effect for suction.

The housing 45 of the suction device 14 shown in FIGS. 14 and 15 together with the cover 44 (not shown here) that closes the housing contains the functions:

- Reception and fastening of the lever 41 of the push-off means 41, - Reception of the lateral, deformable tube 16 of the suction device 14,

- Provision of a passage 47 for receiving and sealing off the outlet of the suction means 15 implemented as suction nozzles, Integration of the compression spring (s) 43, alternatively also by expanding the construction of the tension spring (s),

- Attachment of the part of the fluid connection 16 designed as a suction rod to the rear,

- Shot in a shaft-like cavity.

14 and 15 show that the housing 45 has a receptacle 48 for the armature 42 of the electromagnet 18 and the other components of the forcing means 17 and the fluid connection 16.

FIGS. 16 to 19 show the suction means 15 of a device according to the invention, implemented as suction nozzles.

The subcomponent of the suction means 15 designed as a suction nozzle or suction lip represents the last subcomponent of the entire suction device, consisting of

- vacuum device,

- if necessary reagent collecting vessel (s),

- transversely lying flaup tube 16,

- Forcing means 17 on the lateral, deformable suction tubes 16 or hoses 16 over the suction means 15 or suction nozzle with the reagents to be sucked off.

In order to ensure a safe and reliable, complete suction of the reagents from the reagent gap, a special design of the suction nozzle 15 is required. In particular, three formats of such a suction nozzle are conceivable 15:

According to a first embodiment, the used reagents are sucked off in the middle of the slide 8, which is only raised but not rotated about the longitudinal axis, and in a second version over a corner of the slide 8. This second version is already described in DE 10 2016 225 885 A1, where express reference is made to the relevant disclosure thereof, in particular paragraphs [0077], [0078] and [0084] as well as FIG. 1, and these form part of the disclosure provided here. The first embodiment A shown in FIGS. 16 to 19 relating to a central suction nozzle 15 has a gap 49 at the lower end, which can be coupled to a vacuum device via the connection opening 63 and a further fluid connection 16 and thus the vacuum suction to the end of the Slide 8 diverts so that the reagents can be completely sucked out of the opened reagent gap in an angled state. The complete suction of a used reagent is the decisive feature, because this is the only way to ensure that there is no mixing with the subsequent reagent. To ensure this, the following design and process features are particularly advantageous:

A step lip 50 should touch the back of the specimen slide 8 lying on the two spacers 27, 29 at about 0.1 mm fleas. The total fleas from the bottom of a folding device 7 and the upper surface of a slide 8 is preferably about 1.1 mm, defined by the fleas of the spacers 27, 29 and the thickness of the slide 8. A stepped lip 50, which is directed forwards, for example approximately 1-3 mm long, protrudes over the rear side of the specimen slide 8 and touches its upper side slightly. This direct leaning of the front wall of the step lip 50 on the back of the slide 8 creates a tight connection through which the vacuum suction is passed directly to the reagent gap. The step lip 50 does not touch the bottom of the folding device 7, but ends at a distance from the bottom of the folding device 7 of preferably at least 0.1 mm fleas and thus corresponds to the fleas of the spacers 27, 29 and in particular a maximum of 1.0 mm above the floor of the Folding device 7, ie approx. 0.1 mm before the upper edge of the slide 8 ends on its rear side. A normally standardized slide 8 has a thickness of 1.0 mm, so that the entire flea consisting of spacer 27, 29 and the top of slide 8 is in particular 1.1 mm.

The part of the fluid connection 16 into which the reagents are sucked in by the step lip 50 can be clearly seen in FIG. 19 in particular. The fluid connection 16 can be from contact with the outlet of a connection port 63 to the outlet of the fluid connection 16 consist of a single material, for example a specially designed silicone tube composite or, for example, a deformable piece of tubing that can be deformed by the push-off means 17 and a transverse tube made, for example, of a plastic material with an outlet for the suction means 15 The version of the fluid connection 16, which consists of only one material, has the advantage that a particularly stable and clean component is used which is not restricted in its effectiveness by possible leaks caused by imprecise contacts at the connection points of, for example, two materials (silicone and hard plastic).

An alternative embodiment B of a suction means 15 is shown in FIGS. 29 to 31. In contrast to embodiment A (FIGS. 16 to 19), this does not have a step lip 50 with a gap 49, but an at least substantially round suction opening 62. The suction opening 62 can be coupled to a suction device via the connection opening 63 and a fluid connection 16. The suction opening 62 can be formed in the suction means 15 or can be formed, for example, by the end of a hose, for example a silicone hose. The suction from the gap between the folding device 7 and the slide 8 works extremely reliably if the suction opening 61 has an outside diameter of 5 mm ± 2 mm and an inside diameter in the range from 2 mm to 4 mm, preferably from 3 mm to 3.5 mm . In a further advantageous manner, the suction opening 62 is placed on the specimen slide 8 with no more than half its internal diameter, so that at least the other half of the suction opening 62 remains “free” behind the specimen slide 8. The suction created by the vacuum described above "pulls" the used reagents out of the reagent gap in an excellent way.

Furthermore, the suction means 15 or suction nozzles shown in FIGS. 16 to 19 and in FIGS. 29 to 31 can optionally have a filling passage 64 in order, for example, to introduce a liquid through this filling passage 64 into the folding device 7. 20 and 21 show, in a schematic representation, a perspective view of the first plate 9 of a console 5 of a device according to the invention. It is pointed out once again that the first plate 9 and the second plate 10 can be formed in one piece.

In the following, the first plate 9 is referred to as the base plate 9 and is used to fasten several technical components. A base plate 9 is assigned to each folding device 7. An electromagnet 18, the function of which is to open and close the hose connection 16 of the suction device 14, an electric motor 12 with a plunger 51, which is required for raising and lowering the specimen slide 8, the control means 19 and an attachment option for the temperature control device are located on the underside of this 23 attached to the upper side of the base plate 9. It is essential that a positioning aid 52 is formed on the upper side of the base plate 9 and engages with a corresponding geometry on the cassette bottom 2 so that the two elements are positioned in alignment with one another.

At one end of the base plate 9, the rear end, the housing 45 of the suction device 14 is attached to the upper side of the base plate 9 by the armature 42 of the electromagnet 18 pressing the push-off means 40, which continuously compresses the deformable hose 16, and only opens it for suction when necessary becomes. The temperature control device 23 is attached in the center of the upper side of the base plate 9, and at its other end, the front end, the end of the plunger 51 of the motor 12 with stepped rocker 54 or another end piece protrudes. On the underside of the base plate 9 shown in FIG. 21, magnets 20 are provided which interact with the magnets 20 of the cassette bottom 2.

The cassette bottom 2 can be arranged on both sides of the temperature control device 23 and in the further front area of the base plates 9.

FIG. 22 shows, in a schematic representation, a perspective view of a deflection device 13 and a deflection device 13 with an attached slide 8 of a device according to the invention. A slide 8 lies in the resting state on the spacer 27, 29 resting parallel to the floor in a holding device 7. In the, for example, approximately 0.1 mm high gap between the top of the bottom of the holding device 7 and the underside of the slide 8 with the biological attached Material (tissue section, cells, smears, etc.) contains the reagents necessary for processing, which must be exchanged depending on the procedure protocol. For an accelerated biochemical reaction it can be advantageous to mix a reagent in the gap. This can be done by raising and lowering the slide 8 without suction. For the exchange of a used reagent with a new reagent, the slide 8 is raised and, if it remains briefly in the raised position, is sucked off by activating the suction device 14. The new reagent is then pipetted back into the holding device 7 and the specimen slide 8 is lowered again. Alternatively, pipetting in can also be carried out in lowered, parallel to the spacers 27, 29 of the bottom of the holding device 7, since the capillary forces in the capillary gap created by the lowering sucks in the reagents, regardless of the pipetting point around a slide 8.

In the present invention it is of essential importance that a specimen slide 8 can be lowered completely parallel from the “angled position” onto the spacers 27, 29 of the bottom of the holding device 7. A special deflection device 13 is therefore necessary for raising and lowering, which guarantees that a specimen slide 8 is lowered precisely with its own weight of just a few grams.

The deflection device 13 shown in FIGS. 22 and 23 has the special feature of protruding slightly from the side wall of the cassette bottom 2 in the receiving area 6 in order to prevent the front edge 53 of the specimen slide 8 from sticking to the frame wall. The force for moving the specimen slide 8 (lifting, lowering, rotating) can be generated by an electric motor 12, an electromagnet, an air or water pressure device, a hydraulic device, a spring with a return device, etc. In FIG. 22, the tappet 51 of a motor 12 can be seen, on which a step rocker 54 is pivotably arranged. The step rocker 54 touches the slide 8, lifts this and tilts forward. The slide 8 is thus pressed in the direction of the suction means 15.

The above-described effect that the step rocker 54 can be inclined in the direction of the suction side can be seen in FIG. 23. For this purpose, it is advantageous if the inner wall 58 of the cassette base 2 is designed to be inclined at the points at which the step rocker "rubs" due to the extension of the plunger 51. A corresponding configuration of the cassette bottom 2 is shown in FIG. The bevels 59 to support the movement of the step rocker 54 can be clearly seen therein.

FIG. 25 shows, in a schematic representation, a perspective view of a receptacle 55 temperature control device 23 from obliquely below, the magnets 20 for connection to the console 5 being visible. The receptacle 55 has a cavity 56 for the temperature control device 23 and openings 57 for fastening means, via which the receptacle 55 can be attached to the base plate 9. The temperature control device 23 can be designed differently. The following are conceivable and applicable: a) A small aluminum plate with a resistance heater attached to it, which is passively cooled by the ambient temperature. b) An aluminum plate with a resistance heater attached, which is cooled by active air cooling, for example with an air flow that is directed past the outside of the aluminum plate and, for the sake of simplicity, is taken from the air that is normally already cooled in the laboratory. c) A Termo electric module (Peltier element) that actively heats and cools. d) Water heating and water cooling.

In FIG. 26, the cassette 1 shown in FIG. 3 is shown again. The second positioning hooks 61, with which the cassette 1 can be fastened to the bracket 5 in a form-fitting and force-fitting manner, can be clearly seen. It should be noted that regardless of the specific design of the cassette 1, the overall concept of the invention presented here includes three components within the scope of ease of use, namely the first plate 10 (also called the base plate), the cassette bottom 2 and the cassette lid 3, the " arrange yourself ". This is, firstly, a precise positioning of the removable component, ie the cassette 1 on a defined number of individual base plates 9 and, secondly, the preferably magnetic closure of the cassette bottom 2 and the cassette lid 3 to form a cassette 1. Looking at these three components, individual base plates 9, cassette bottom 2 and cassette lid 3 from bottom to top, it becomes clear that one or more permanent magnets 20, 21 are embedded in the three components, which are directly above one another for optimal use of the magnetic forces are appropriate. The cassette bottom 2 is equipped with permanent magnets 20, 21 at two points. To form the cassette 1, the magnets 20 are attached in the three longitudinal struts 24 both in the cassette bottom 2 and in the cassette lid 3. The magnets 21 for locking the cassette bottom 2 or the entire cassette 1 are let in next to the plunger 51 behind the overflow channel 11. The complementary magnets 21 are embedded in the base plates 9 at the same point, that is, next to the plunger 51. The user therefore only has to move a cassette 1 into the “relative proximity” of the intended area of the console 5 in which the cassette 1 is to be used, which comprises several base plates 9. Relative proximity is to be understood as meaning, for example, a distance of 10 mm to approx. 20 mm from the magnets 20 of the base plates.

In FIG. 32, an exemplary embodiment of a method according to the invention is shown on the basis of a block diagram.

In a first step 65, the specimen carriers 8 carrying the biological material are each placed on a holding device 7 before or after the holding devices 7 have been inserted into the receiving areas 6 of the cassette base 2. In a second step 66, the cassette lid 2 and the cassette base 3 are brought together in order to thus close the cassette 1. Then, in a third step 67, the cassette 1 is connected to the console 5. In a fourth step 68, the liquid or the Liquids introduced into the holding devices 7. If necessary, the samples are then processed further, for example the slide 8 is lifted and the liquid is sucked out of the reagent gap, after a defined exposure time.

It should be noted that further steps can be carried out between the individual steps 65 to 68, for example heating the slide. Furthermore, steps 65 to 68 do not necessarily have to take place in the specified sequence.

With regard to further advantageous embodiments of the device according to the invention, reference is made to the general part of the description and to the appended claims in order to avoid repetition. Finally, it should be expressly pointed out that the above-described exemplary embodiments of the device according to the invention are only used to discuss the teaching claimed, but do not restrict them to the exemplary embodiments.

List of reference symbols

1 cassette

2 cassette bottom

3 cassette covers

4 cover disc

5 console

6 Recording area

7 holding device

8 microscope slides

9 first plate 0 second plate 1 overflow channel 2 electric motor 3 deflection device 4 suction device 5 suction means 6 fluid connection 7 pressure means 8 electromagnet 9 control means 0 magnet 1 magnet 2 side wing 3 temperature control device 4 longitudinal strut 5 spacer 6 side wall 7 support shoulder 8 rear wall 9 support point 0 tear-off edge 1 bulge indentation

Slope

Pressure point

deepening

indentation

Lid strut

engine

bracket

Abutment

lever

anchor

feather

cover

casing

head Start

Passage

admission

gap

Step lip

Plunger

Positioning aid

Edge

Step seesaw

admission

cavity

openings

Inner wall

Inclined first positioning hooks, second positioning hooks, suction opening, connecting opening, filling passage, first step 66 second step

67 third step

68 fourth step

Claims

Expectations

1. A device for wetting biological material with at least one liquid, comprising a cassette (1) with a cassette bottom (2) and a cassette lid (3), the cassette (1) having at least two receiving areas (6) each for a holding device (7) ), wherein the holding device (7) serves to receive a slide (8) which carries the biological material, the biological material arranged on the slide (8) being wettable with the liquid within the closed cassette (1) and wherein the The slide (8) within the closed cassette (1) can at least partially be lifted off the holding device (7) in order to enable the liquid located between the slide (8) and the holding device (7) to drain off.

2. Device according to claim 1, characterized in that the holding devices (7) can be removed from the cassette (1).

3. Device according to claim 1 or 2, characterized in that at least part of the receiving area (6) is formed by a passage in the cassette base (2).

4. Device according to one of claims 1 to 3, characterized in that a console (5) is arranged, wherein the cassette base (2) can be placed on the console (5) and removed from it.

5. The device according to claim 4, characterized in that the console (5) has a deflection device (13) so that each slide (8), preferably individually, can be raised within the closed cassette (1).

6. Apparatus according to claim 4 or 5, characterized in that a temperature control device (23) is arranged on the cassette (1) and / or on the console (5) for each receiving area (6).

7. Device according to one of claims 1 to 6, characterized in that the cassette lid (3) and the cassette bottom (2) can be connected to one another via magnetic forces, in particular via at least one permanent magnet (21).

8. Device according to one of claims 1 to 7, characterized in that the cassette lid (3) and the cassette bottom (2) form-fitting and / or force-fitting, in particular via a clamping and / or latching mechanism (60), for example via a bayonet lock, are connectable to each other.

9. Device according to one of claims 4 to 8, characterized in that the cassette (1) and the console (5) can be connected to one another via magnetic forces, in particular via at least one permanent magnet (20).

10. Device according to one of claims 4 to 9, characterized in that the cassette (1) and the console (5) positively and / or non-positively, in particular via a clamping and / or latching mechanism (61), for example via a bayonet lock , are connectable to each other.

11. Device according to one of claims 1 to 10, characterized in that a suction device (14) is designed for sucking off the liquid from the holding device (7).

12. Device according to one of claims 1 to 10, characterized in that the suction device (14) has suction means (15) which are arranged on the cassette lid (3) or on the cassette base (2) in order to remove liquid from the holding devices (7 ) suction.

13. The device according to claim 12, characterized in that the suction means (15) are connected to a vacuum device, for example a pump, via a, preferably common, fluid connection (16).

14. The device according to claim 13, characterized in that the

Fluid connection (16) is at least partially or entirely designed as a hose connection.

15. Device according to one of claims 12 to 14, characterized in that pressure means (17) are provided in order to interrupt the fluid connection (16) of a suction means (15) with the vacuum device.

16. The device according to claim 15, characterized in that the

Forcing means (17) is formed by a pretensioned forcing element (41).

17. The device according to claim 16, characterized in that the

Forcing means (17) can be moved against its bias by a release device (18), for example an electromagnet or an electric motor, in order to release the fluid connection (16).

18. Device according to one of claims 1 to 17, characterized in that side wings (22) are formed on the holding device (7) and that indentations (32) are formed on the receiving area (6) on which the side wings (22) rest .

19. The device according to claim 18, characterized in that the depth of the indentations (32) is greater than the thickness of the side wings (22).

20. Device according to one of claims 1 to 19, characterized in that pressure points (34) are arranged on the cassette cover (2) in order to press the holding devices (7) in the direction of the cassette bottom (2).

21. A method for wetting biological material with at least one liquid, using a device according to one of claims 1 to 20, with the following method steps:

- Insertion of slides (8) carrying the biological material onto a respective holding device (7) before or after the holding devices (7) have been inserted into the receiving areas (6) of the cassette base (2),

- bringing together the cassette lid (3) and the cassette base (2) to close the cassette (1), - connecting the cassette (1) to the console (5), and

- Introducing the liquid and / or the liquids into the folding devices (7).