DE102009015596A1 - Method and device for the treatment of carrier-fixed material - Google Patents

Abstract

The invention relates to methods and apparatus for the treatment of carrier-fixed material, in particular biological material or material interacting with biological material. In one aspect, the invention provides a method of treating carrier-fixed material, particularly biological material or material interacting with biological material, using a plurality of slides arranged vertically above one another in a first carrier cassette, on each of which the material is disposed, and wherein a) in each case a slide is at least partially removed from the first carrier cassette, b) at least one treatment agent is applied to the material on the at least partially removed slide, c) the slide is placed in the first or a second carrier cassette after application of the treatment agent and d) the material on the slide in the first or the second carrier cassette is incubated with the treatment agent, and wherein at least one of the steps a) to c) is carried out automatically. In a further aspect, the invention provides a method in which on the slide manually, semi-automatically or automatically delimited by means of a limiting means a treatment area and the material required or appropriate for treating the material amount of treatment agent automatically on the basis of the material on the ...

Description

The This invention relates to methods and apparatus for treatment of carrier-fixed material, in particular biological Material or material interacting with biological material.

Background and state of the technology

The Treatment and in particular staining of cell or tissue samples has long been an important part of the medical Diagnostics. Accordingly, a whole series of treatment or staining method. These include in particular immunohistochemistry or immunofluorescence, the chromogenic or fluorescent in situ hybridization and the special stains with dyes. A modification of these methods in recent Time through the microarray technology.

at The various treatment / staining procedures are carried out regularly the fixation of an examination subject (eg a tissue section) on a microscope slide and the treatment or staining of the examination subject with different reagents. The microarray technology fixes reagents (eg, proteins such as antibodies, or nucleic acids such as oligonucleotides) on the slide and staining with a subject. The Treatments / stains often include a large sequence of incubation and washing steps, which eventually led to staining of the object under investigation.

For some of the treatment / staining procedures are commercial Automation solutions available. One of the first available automation solutions works after the capillary gap method. This will be slides vertically arranged so that between each two slides a capillary gap is created. The immersion of the head of the slides into a reagent causes the gap to move against gravity filled with reagent. The reagent will be according to the desired Incubation time removed by suction pads from the gap. It will be through the vending machine groups of slides of one Reagent Reservoir moves to the next one.

In currently available staining machines, which are also referred to in particular in connection with the immunohistochemistry as "autostainer", usually horizontally and stationarily arranged slides by means of a movable pipetting head reagent and washing solution is supplied according to a computer-controlled staining protocol. Such staining machines are for example in the WO 99/49295 . WO 2004/074847 or WO 2004/058404 described. The protocol is defined by the user and assigned directly to a slide or defined by a code located on the slide and processed accordingly.

An alternative solution is in the WO 99/04432 described. Here, slides and reagent containers are each arranged on a carousel. By a relative movement of the two carousels, slide and reagent supply are arranged for the respective current staining step so that the reagent can be applied from a cartridge to the slide. The slides are arranged horizontally next to each other on the carousel.

In the US 2003/0099580 A1 a cassette is described in which slides are arranged one above the other. The cassette should be usable in staining machines. However, it is not specified how this can be done. The cassette is equipped with drip holes and can be used for the simultaneous staining of slide bundles in staining trays, as are commonly used in histology. A coloring of individual slides from the cassette is not provided. In the WO 99/34190 an array of vertically stacked, slide containing cartridges is described. The elaborately designed cassettes should also be usable in automated sample treatment devices. However, here too there is no corresponding information as to how this should be done.

Previous automation solutions have various disadvantages. A disadvantage of the known solutions is, for example, that a relatively high space requirement exists for a limited number of slides. With a high dye yield, the limited capacity thus leads to the necessity of several parallel machines to increased costs and again increased space requirements. At the same time, especially in the WO 99/04432 described construction to comparatively long sample processing times.

Another disadvantage is the limited access to a dyeing run. Once a run has been started, it must first be stopped before further slides can be subjected to a dyeing process. To mitigate this drawback, it is known in the art to add a number of slides to a dyeing run for extra free space provided therefor. Furthermore, it is known from the prior art to group slides on drawers. As soon as a drawer has been processed, the slides can be removed and others can be brought into the machine with the drawer. A continuous However, feeding individual slides and their integration into existing dyeing runs is not possible.

A particular disadvantage of the previous methods is the high reagent consumption, which is due to the fact that whole slides or large parts thereof must be wetted with reagent. The WO / 2000/008511 proposes a solution to divide slides into three zones separated by hydrophobic barriers before applying a sample. This solution has the disadvantage that the examination subjects must be positioned accurately and the reagent consumption can not be adjusted individually. In the WO 04/001389 a special slide cover is proposed, with the help of a reaction chamber is formed, the size minimization should also lead to a minimization of the required amount of reagents. Even with this solution, however, costly reagents are also spent on areas that contain no object of investigation. In addition, this solution is technically very complex to realize, since the cover must be moved for each reagent change.

task The present invention is the automated and continuous Treatment of a biological sample while avoiding the disadvantages of State of the art to allow. Furthermore It is an object of the present invention to reduce reagent consumption to minimize in automated sample handling.

The Tasks are solved by the objects of the sibling claims.

• a) jeweils ein Objektträger zumindest teilweise der ersten Trägerkassette entnommen wird,
• b) mindestens ein Behandlungsmittel auf das Material auf dem zumindest teilweise entnommenen Objektträger aufgebracht wird,
• c) der Objektträger nach dem Aufbringen des Behandlungsmittels in der ersten oder einer zweiten Trägerkassette abgelegt wird, und
• d) das Material auf dem Objektträger in der ersten oder der zweiten Trägerkassette mit dem Behandlungsmittel inkubiert wird,

und wobei mindestens einer der Schritte a) bis c) automatisiert vorgenommen wird.In a first aspect, the invention provides a method of treating a supported material, particularly a biological material or material interacting with biological material, using a plurality of slides arranged vertically above one another in a first support cassette, on each of which the material is applied, and where

• a) each slide is at least partially removed from the first carrier cassette,
• b) at least one treatment agent is applied to the material on the at least partially removed slide,
• c) the object carrier is deposited in the first or a second carrier cassette after the application of the treatment agent, and
• d) the material on the slide in the first or the second carrier cassette is incubated with the treatment agent,

and wherein at least one of the steps a) to c) is carried out automatically.

The inventive method allows a partially and even fully automatic material treatment. Under A "material treatment" is used in the present In particular the treatment of biological material, for example cells, tissue sections, tissue punches, microarrays or the like understood by dyeing. The term "material" includes also material that is not of biological origin, but with Material of biological origin can interact, z. B. synthetic oligonucleotides. "Biological material" is any material of biological origin, d. H. one in a living being naturally occurring or produced by a living being Material, such as cells, tissues, cell extracts, cell lysates, Cell smears, serum, proteins, nucleic acids (eg RNA, DNA), antibodies, lectins and the like. Under a "with biological material of interactive material "becomes Material understood that itself not necessary of biological origin but preferably with material of biological origin specific interaction is received. An example of such Material is a synthetic oligonucleotide. Such with biological Material interacting material, even of biological origin or not, is also referred to as "probe" here. The term "coloring" includes here any Marking a material or a material component by a directly or indirectly visually perceptible means, for example a dye, an antibody or a nucleic acid probe, and therefore includes, for example, both conventional histological staining as well as immunostaining, in situ hybridization and Microarray techniques. Examples of conventional histological Dyeings are known to the person skilled in the art and include, for example hematoxylin-eosin stain, Gram stain, the Giemsa stain, the toluidine stain etc. The application of the material to a microscope slide as well its fixation on the slide is not intended by the Be covered by the term "material treatment", wherein the term "fixation" also the preservation a biological sample. The term "material treatment" includes also not the embedding of a sample, for example in paraffin. He concludes, however, if necessary, the dewaxing a Sample.

Under a slide is here understood any carrier on which a material can be fixed according to the above definition. It may, for example, be a glass slide in the form of a glass plate, as commonly used in microscopy or histology, for example. Such a slide preferably has the according to ISO 8255-2 usual dimensions (26 × 76 mm × 1-1.5 mm), but can also have other dimensions. It may also be a plastic slide, a membrane or the like act, provided that the carriers have sufficient stability. "Carrier fixed" or "fixed on a support" here means that a material, for. As a sample or a probe is applied to a slide, wherein the material preferably does not solve by merely tilting or rotating the slide due to the pure gravity effect of this again. The material is preferably applied to only one of the surfaces of the slide.

Under a "sample" is understood here as any material this, in contrast to the "probe" itself subject of investigation is. Under a "biological sample" becomes a Sample understood from biological material, for example a Cell or tissue sample, in particular a flat cell or tissue sample, for example a tissue section or a cell smear. It can be z. B. a sample of plant, animal or human Act of origin. Preferably, the sample is a human sample Cells or human tissue. If, in the following description, in particular in the embodiments, the invention with reference the use of a "biological sample", e.g. B. a tissue sample is explained in more detail means This does not mean that a limitation is intended is. Rather, the skilled person will recognize that the example of a also describes such methods or devices are applicable to material that is not biological Sample in the narrower sense defined above, but a "material" according to the above Definition, e.g. B. a probe. So if the treatment is on a slide fixed biological sample described will be readily apparent to those skilled in the art that the embodiments according to the microarray technology, z. B. DNA microarrays or protein microarrays, are applicable, in which, unlike classical histological techniques, the actual object of investigation, the commonly used as a "sample" is present, not fixed on the slide, but is added to a carrier-fixed probe, the biological Origin (such as in the case of antibodies) or not (as in the case of synthetic oligonucleotides), however, interacts with biological material, and in the case of the sample or sample components are often labeled. For example can in a DNA microarray a cDNA on the slide fixed and the actual object of investigation, for example a cell lysate with labeled DNA molecules linked to the cDNA interact, in this case can hybridize is placed on the slide to a staining reaction cause.

In another example may be an antigen on the slide be fixed and serum is added.

As well is, if here for the sake of better understanding, exemplary from "sample treatment" is spoken, so no restriction on the treatment of slides intended fixed samples in the above narrow sense, if not expressly stated otherwise. Much more the person skilled in the art will recognize that the corresponding embodiments, even if in it only by "sample treatment" or "treatment a sample "is mentioned, also for example on the Miroarray technology are applicable, and the term "sample treatment" or appropriate terms, unless expressly stated otherwise stated, the treatment of a carrier-fixed Includes materials, such as a probe.

The method according to the invention is not on the treatment of only one material, z. B. a tissue section on limited to a slide. Rather, it is also possible, several materials, eg. B. biological samples like a tissue section, spatially separated on a single To accommodate slides, it being preferred, though it is not necessary for the materials to be on a microscope slide each treated the same.

Under "automated" or "automatic" is in the present application understood that the corresponding process step or the process steps not manually by a person, but completely from a machine, preferably computer-controlled, is or will be carried out. The term "semi-automatic" means here, in an otherwise automatic process at least one step manually, d. H. by a person. For example, at the beginning, at the end or in the course of a series a manual step is provided by automatically running steps be, for example, made by a person or a selection a reagent is pipetted. Under a "semi-automatic" procedure Thus, a method is understood that at least one automatically has expired and at least one manually made step, in the case of multiple automatic and / or manual steps does not matter what proportion of the number have the manual steps to the whole process. This here term "manual" also includes one of a person's act involving a technical device is used.

By a "treating agent" is meant here any means that can be applied to a material, e.g. B. a tissue sample is applied in order to achieve a certain effect there, for example, a means for dyeing the material or a means for Removal of a dye or excess dye. The term therefore includes, for example, coloring or coloring reagents, but also antibodies or nucleic acids, preferably in solution, the z. B. in immunohistochemistry or in situ hybridization can be used, and auxiliary reagents such. As aqueous or organic solutions for pre- and post-treatment, such as buffers and / or washing solutions. However, the term includes, for. As in the case of microarrays, and samples or sample components that may be colored or otherwise marked. It may therefore z. As well as a cell lysate, serum, cells or tissue. As a rule, the treatment agents are available as aqueous or organic solution. The term "treating agent" is used synonymously with the term "reagent" and includes, for example, mixtures or combinations of various agents.

at Previous methods are individually colored materials or samples arranged on slides that a relative have a mutually fixed position in a horizontal plane, to treatment agent by, for example, a pipetting device supply. It has been surprisingly found that such an arrangement in terms of an efficient, cost-effective and automatable material or sample treatment is not required is, and at the same time a space-saving, fast and individual Processing of slides is enabled, if you put the slides vertically one above the other arranges, for example in a suitably designed carrier cassette. "Vertically superimposed" means here that the material bearing surfaces of several Slides each in the horizontal plane, d. H. perpendicular to the direction of gravity, and beyond along the direction of gravity on each other are arranged. In the case of flat slides, z. B. conventional microscope slides, means "vertically stacked" example, that the individual slides with their longitudinal axis essentially horizontally and with the material-loaded surfaces are oriented opposite to the direction of gravity, and several such slides are substantially perpendicular to each other, d. H. are arranged in the direction along the direction of gravity. The term "vertically superimposed" should describe here in particular slide assemblies, in which the slides are aligned with their sides to each other arranged, d. H. without significant horizontal misalignment, in the Usually, however, vertically spaced from each other, one above the other but are also covered arrangements in which the wearer are arranged offset from each other in the horizontal, wherein however, the offset is preferably as small as possible.

at the inventive method is at least one of the steps a) to d) carried out automatically. Preferably the steps a) and c) are automated while Step b), for example, manually. Particularly preferred however, also step b) is done automatically, so the steps a), b) and c) are automated. At the incubation step d) As a rule, neither manual nor automatic intervention takes place.

Prefers is an embodiment of the method in which e) the treatment agent is removed after incubation.

For example The treatment agent can be washed off with water or a buffer become. The treatment agent may also be in other ways, for. B. by compressed air or by simply pouring removed become.

The Treatment or coloring of materials often involves an alternating sequence of treatment and washing steps. Be applied to a slide materials with a treatment agent, e.g. B. a dye solution, a Enzyme solution or the like, overcoated, then incubated for a certain period of time, then washed, to remove residues of the treatment agent, and with another or optionally again overlaid with the same treatment agent. In the method erfindungsge MAESSEN these Steps, for example after specification of a specific treatment protocol, preferably carried out automatically. The steps a) to e) are therefore preferably repeated until the material on the slide completely treated, z. B. dyed, is.

Prefers is in the inventive method to the Treatment of the material required or appropriate Amount of treatment agent automatically determined. Especially preferred this is done on the basis of the material, such. B. a Tissue section on which slide takes. this makes possible it, the necessary or appropriate for the treatment of the material Need to identify treatment agent and the use of the treatment agent automatically limit to this level. This allows the Treatment agent consumption can be reduced to a minimum.

In a particularly preferred embodiment of the method according to the invention is on the slide manually, semi-automatically or automatically, z. B. delimited by a limiting means, a treatment area, wherein the treatment area the material, for. A tissue section or an oligonucleotide array. For example, in the case of an oligonucleotide array, the treatment area includes the area on the Slides in which the oligonucleotide probes are fixed. The treatment area can be predetermined by markings on the slide and can be detected by means of the markings, for example by means of optical, optoelectronic or other methods. In the case of a biological sample, z. As a tissue section, the treatment area comprises the area that the sample, z. B. the tissue section, on the slide occupies. In this way, the area of the slide containing the material can be separated from the rest of the slide. The area referred to as the "treatment area" or optionally also as the "reaction area" or "reaction zone" is applied to the material, optionally with the aid of the above-described markings, on the slide, on or in the treatment agent, for example reagents become. Thus, not the entire slide or an unnecessarily large portion of the slide need be covered with reagent, but essentially only the area containing the material, resulting in significant reagent savings.

Of the Term "material area" refers to the Area or area in which or on the material is on the slide.

The above mentioned markers with which the slide can be provided before or after the application of the material, mark the material area. They preferably mark border points an area containing the material. The of The marked areas can, but must not to be identical with the treatment area.

In In a second aspect, the invention relates to a method of treatment a material applied to a slide, in particular biological material or interacting with biological material Materials, where on the slide manually, semi-automatic or automatically, for example by means of a limiting means or by drying a slide area, a the material delimited and the treatment of the Material required or appropriate amount at treatment agent automatically on the basis of the material the slide occupied surface, based markings on the slide characterizing this area or based on the area of the treatment area and is dosed.

The Method according to the second aspect of the invention allows the use of treatment agents, for example to control in medical diagnostics or histology and / or to minimize. The procedure is both individual and applicable to several material loaded slides and can be completely automated if necessary. The Arrangement of the slide is arbitrary. Several Slides can thus be next to each other horizontally, vertically stacked or otherwise arranged. The treatment of the located on the slide Material used amount of treatment can with the help of inventive method to the required or appropriate, d. H. on the overlooking the desired treatment result necessary or appropriate Measure be limited.

• i) das automatische Erfassen des Materials und/oder von den Materialbereich kennzeichnenden Markierungen auf dem Objektträger,
• ii) das automatische Ermitteln der auf dem Objektträger von dem Material eingenommenen oder der von den Markierungen gekennzeichneten Fläche,
• iii) das automatische Abgrenzen eines Behandlungsbereichs auf dem Objektträger um die von dem Material eingenommene und/oder die von den Markierungen gekennzeichnete Fläche,
• iv) das automatische Ermitteln der zur Behandlung des Materials erforderlichen oder zweckmäßigen Menge an Behandlungsmittel, und
• v) das automatische Aufbringen der ermittelten Behandlungsmittelmenge innerhalb des Behandlungsbereichs auf den Objektträger.

In a preferred embodiment, the inventive methods according to the first and second aspects of the invention comprise the following steps:

• i) the automatic detection of the material and / or markings on the slide characterizing the material region,
• ii) automatically determining the area occupied by the material on the slide or marked by the markings,
• iii) automatically delimiting a treatment area on the slide around the area occupied by the material and / or the area marked by the markers,
• iv) automatically determining the amount of treatment agent necessary or appropriate to treat the material, and
• v) automatically applying the determined amount of treatment agent within the treatment area to the slide.

The Delineation of the treatment area on the slide can, for example, by applying a limiting means the slide or by drying the slide surface those taken from the material and / or from the markings marked area done. To delineate a treatment area the slide comes next to the application of a limiting means, z. As an oil, wax, plastic or the like, also the drying of the corresponding areas or areas in question around the material. This can be done with help, for example an acetone pin, blower or the like happen. The removal of itself on a slide often, z. B. generated by washing steps, forming moisture film also a barrier to the aqueous solutions be withheld. Of course you have to not all located outside the treatment area Area of the slide treated in this way, d. H. covered with a limiting agent or freed of moisture become. It suffices a barrier, for example in shape a contour line, z. B. with a thickness of 0.5-5 mm, which can be interrupted by the way.

the It is obvious to a person skilled in the art that steps i) to iv) generally, but not necessarily, for each slide or each material must be performed only once. Once determined, the amount of treatment agent changes in the course of treatment, which regularly includes several treatment steps, z. As coloring steps, includes, not normally, so that the different treatment agents that make up one material sequentially treated, applied in the same, once determined amount become. It may, depending on the type of treatment and the used Be necessary to repeat step iii), to maintain the barrier function.

ever However, it can also be useful to use it once determined amount of treatment agent in the course of the treatment of a slide to vary, d. H. to increase or to degrade, for example, depending on the treatment agent or depending from the reached treatment stage. In a corresponding application could in a variant an already applied limiting means initially removed in an additional step and with greater or lesser distance to the edge of the material reapplied. This variant would only required, in addition to the additional step of Removing the treatment agent, the repetition of the above steps iii) and iv). In another variant could the amount of treatment agent based on changed specifications simply be redetermined and changed without that Limiting means must be removed. This would only be the repetition of step iv) is required, the determination the amount of treatment agent with changed investigation specifications would be done. Also any combination of both Varian is possible. Although not preferred, the treating agent may also be in excess, i. H. in a more than required amount, to be added, for example in the case of buffer and / or wash solutions.

Of the future treatment area is automatically detected by using the material on the slide with appropriate Means, such as an image processing system, automatically recorded and automatically determines the area that the material occupies the slide. This can too by doing that marks on the slide intended to identify the area containing the material contains, are recorded and based on these markings the area to be treated is determined. This is then advantageous if the material itself is not or only with difficulty the slide can be detected. The detection The material on the slide may optionally also with the help of sensors, z. B. turbidity sensors, scattered light sensors and the like, be made or supported. Subsequently the treatment area on the slide through automatic application of a limiting agent to the slide or by drying the slide surface around the area occupied by the material and / or of area marked as marked. The actual Treatment area, d. H. the area where that later added treating agent is to act, is limited. The necessary or expedient for the treatment of the material Amount of treatment agent is then automatically determined, wherein preferably the surface of the generated Treatment area is used. The determined amount of treatment agent is then automatically within the treatment area on the Slide applied. Since the treatment area the comprises the area occupied by the material, the Material covered in this way by the treatment agent.

By The automatic material area recognition can be customized Reaction zones are generated on the slide, the preferably include the material area as closely as possible. The treatment area may be larger than that of the material the slide or markers marked area, but preferably corresponds to Essentially that of the material on the slide occupied or marked by marks area. This makes it possible to add only the amount of treatment use that is actually needed to that Treat material, for example, to color. Even though preferred, it is not necessary that the course of the limiting means or the drying zone formed by the boundary exactly follows the edge of the material or corresponds. Especially for materials with irregular Edge may be appropriate, here a smoothing provide the boundary means limit and the limiting means on an envelope enclosing the irregular edge of the material applied. For materials that are difficult or impossible due to optical or other methods are detected automatically, preferably electronically can, is provided, the treatment area based on determine markings applied to the slide. For example, these marks can be four vertices define a rectangular treatment area. The marks can be present in any number and arrangement, so that different treatment areas can be realized can. The markings may be appropriate also encoded information, eg. As coordinates, location information, size information in formations etc., included. The area bounded by the marks may correspond to that of the treatment area or, if appropriate also be smaller.

The Limiting agent is preferably a hydrophobic agent, as the Treatment of biological material usually with the help of aqueous Solutions done. The hydrophobic agent is preferably a natural or synthetic oil, fat, resin or wax or a plastic. It can also be anything other agent applied to a slide, is able to prevent a treatment agent from a given Area on the slide leaves. It can Depending on the application, it may also be a hydrophilic agent. It is also possible, if necessary, for the limiting means to replace it with another, if any, in the course of treatment of a material may be necessary or appropriate should.

Prefers encloses the limiting means or the dried surface, which is also referred to here as a "dry zone", that occupied by the material on the slide Area completely. "Completely enclose "means here that the limiting means or the drying zone a continuous, d. H. uninterrupted, Bound around the material around. This is not required. The limiting means or the drying zone can do this Even partially enclose material on the slide. For example, in the case of slides as they are commonly used for The microscopy used, possible, limitations in each case apply only to the narrow side of the slide out and the other pages, d. H. the long sides, free to to let. Due to the precisely calculated and dosed amount of treatment takes place with substantially horizontal storage of the slide no running down of the treatment agent on these sides. One Applying limiting agent or a drying zone is attached to the Long sides of the slide therefore dispensable, causing Time and limits can be saved.

The Identification of the treatment of the material on the slide required or appropriate amount of treatment agent is preferably made on the basis of the treatment area. she However, it can also be based on the material on the slide or the area marked by the markings become. This applies both in the case of the treatment area essentially that of the material on the slide occupied or marked by the marks area corresponds to what is preferred as well as in the event that the treatment area is greater than that of the material taken on the slide or the Area marked by the marks.

The Processes according to the invention furthermore preferably vi) automatically controlling the successful application of the treatment agent within the treatment area on the slide. This is advantageous because in this way the possibility is given to determine whether treatment agent has been dosed or Not. This may be the case, for example, when the reagent supply is exhausted or a constipation or another Defect is present, which prevents a treatment agent dosage. In such a case, for example, a warning and / or abort the further processing. The control can example, by means of the above-mentioned image processing system done and z. B. on the determination of optical changes which result from dosing a treatment agent. Other means, e.g. B. conductivity measurements, etc., are of course also possible.

In In a third aspect, the present invention relates to a device for carrying out the inventive Method, wherein provided an electronic image processing system is that designed as an automatic material detection system is.

The For example, an electronic image processing system can be a digital one Camera or other image capture unit and an electronic Include image analysis unit. Preferably, the electronic comprises Image processing system a digital camera and associated hardware and software technically equipped staff Computer (PC). In addition, the electronic image processing system also special sensors, z. B. Turbidity or scattered light sensors include, to capture the material on the slide too enable or facilitate. That of the digital Camera is generated digital image of the slide from the image analysis unit automatically to the presence, the Location and extent of a material, such as a biological Sample such as a tissue section, or optionally for the presence analyzed by the treatment area characteristic markers. The term "automatic material detection" should here also include the case that only slide markings, which mark the material area are automatically detected. From the image data, possibly taking into account encoded information contained in the markings can determine the area that the material, z. As a tissue section, on the slide occupies. From this, the amount of treatment agent can be determined, the Treatment of the material required or appropriate is. The electronic image processing system can also be used determine if any treatment agent and, at all, if so, if sufficient treatment was dosed.

In In a preferred embodiment, the device comprises a limiting agent metering station for automatic application of restricter or dry Surface (dry zone) on the material loaded slide. The delimiter dosing station doses automatically, and preferably computer controlled, a limiting means on the slide or creates a corresponding drying zone to a slide on the slide Create treatment area.

The Begrenzungsmitteldosierstation is preferably so with the electronic Image processing system connected to the Begrenzungsmittososierstation based on data provided by the electronic image processing system be determined, is controllable. The data can do this be used to control the delimiter dosing station so that with the help of the dosed by the Begrenzungsmittosumierstation Limiting a treatment zone on the slide is delimited by the material taken on the slide Includes surface and this preferably as possible tightly encloses.

Prefers the device has an input station, which is configured and arranged to a first carrier cassette to receive, wherein the first carrier cassette designed and is set up that the slides in it vertically one above the other can be arranged. Preferably, the slides are in the Carrier cassette can be arranged flush with one another, d. H. so that they do not or only slightly in the horizontal direction offset from each other. A carrier cassette can for the inclusion of, for example, at least 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 50, 100 or more slides designed be without the space requirement of the device increased significantly becomes.

Further Preferably, a Trägermanipulator is provided, the so designed and set up to be a slide individually and automatically, preferably computer-controlled, the first Removed carrier cassette and a treatment, preferably in a treatment station, can be supplied, and the slide after treatment in the first or a second carrier cassette can be stored.

at the device according to the invention may be the treatment the individual slide or the material fixed thereon each carried out so that a single slide by means of of the carrier manipulator either partially or completely taken from the carrier cassette and the treatment station is supplied. The treatment station, for example one or more reagent containers and one or more May include pipettes, may be stationary or movable, but is preferably stationary. Also a combination stationary and movable components is possible. For example, a component to which the slides be washed while being stationary the components that help the actual coloring takes place, are designed movable. The electronic image processing system and the delimiter dosing station are preferably in the Integrated treatment station. The treatment station can be spatially and / or structurally separate units, each for identical or different treatment steps are responsible. For example, a detergent dosing unit that only Detergent dispensed, and a reagent dosing unit containing only reagents, with the exception of detergent, dosed, be provided. The detergent dosing unit can while stationary while the reagent dosing unit can be mobile. The reagent dosing unit may be, for example be configured as a mobile pipette. The units can spatially and / or structurally separate from or to Be summarized processing groups. It can too several treatment stations be present.

Of the can be treated by the treatment station slides then returned to the carrier cassette again from which it was taken. But it is also possible the slide after a treatment or recognition step, z. B. for incubation of the material with the treatment agent or to form slide groups with the same or similar Way to be treated materials, in another carrier cassette to spend. In one embodiment, therefore, are a first carrier cassette and a second carrier cassette provided, wherein the device is set up and configured is that the slides by means of the carrier manipulator the first carrier cassette taken individually, the Treatment station supplied and then the second carrier cassette can be supplied.

The Number of carrier cassettes simultaneously from the device can not be processed on one or two carrier cassettes limited. Rather, it is intended and preferred that two, three, four, five or more carrier cassettes simultaneously can be processed independently of each other. The supply of the carrier cassettes is preferably done via an input station, whereby the inventive Device allows almost any number of slides at almost any time of a running dyeing process to integrate into the treatment process.

Particularly preferably, the carrier manipulator is electronically controlled. Preferably, the Trägerermanipulator along at least one coordinate axis movable, so that a movement of the slide along at least one coordinate axis can take place. The carrier manipulator can also be designed such that a movement of the object carrier can take place along all three coordinate axes. A coordinate axis is understood to mean the axis of a Cartesian coordinate system with abscissa (x-axis), ordinate (y-axis) and applicate (z-axis). Terms used herein, such as "x-direction", "y-direction" and the like, refer to the respective axes of the coordinate system.

the One skilled in the art will readily appreciate that carrier handling can be done by any mechanism. In a particularly preferred Embodiment, the carrier manipulator is a vacuum pipette, a gripping tweezers or a robotic arm.

the It is also obvious to a person skilled in the art that it is also possible to design the carrier manipulator stationary and for example the carrier cassette with the aid of appropriate Moving transport facilities. Basically, the One skilled in the art will recognize that relative movements, provided they are in the invention are of importance, can be generated by either one component stationary and the other mobile or conversely configured.

The The invention will be described below for the purpose of illustration of figures to preferred embodiments of the invention explained in more detail. Even if in the embodiments, the Treatment, in particular staining of a biological sample is in the foreground, a restriction on this is not intended. Rather, the examples are also applicable to the case, that is not the actual subject of investigation on the slide is fixed, but the appropriate reagent for treatment / staining, such as For example, in the microarray technology. It shows:

1 A simple scheme of an embodiment of the invention.

2 A simplified view of another embodiment of the invention.

3 A simplified view of the embodiment of 2 in a different operating state.

4 A simplified view of the embodiment of 2 and 3 with other components.

5 . 6 A top view of a part of in 4 illustrated embodiment.

7 Another view of part of the in the 2 - 4 illustrated embodiment.

8th A simplified view of another embodiment of the invention.

9 A simplified view of part of another embodiment of the invention.

10 A schematic plan view of a part of the embodiment of 9 ,

1 shows a highly simplified schematic representation of an embodiment of the invention, which can be used for example for the automated immunostaining of tissue sections. Shown is the top view of an arrangement containing a pretreatment module 21 and a treatment module 22 , for example, an immunostaining module. In the pretreatment module 21 For example, dewaxing or dewaxing of sample-loaded slides is done 2 as well as heating tissue sections for antigen recovery, in the treatment module 22 For example, the immunostaining of the samples takes place in several steps. The pretreatment module 21 and the treatment module 22 each comprise an input station 3 one with slides 2 loading carrier cassette 1 (here, unlike the other components, shown in side view), and an output station 4 , From the entrance station 3 to the starting station 4 go through the slides 2 various pretreatments. In the pretreatment module 21 At various stages, the embedding medium, e.g. As paraffin removed. It can be different bathrooms 5 act in which the carrier cassette 1 with the slides 2 immersed in succession. The carrier cassette 1 goes through the pretreatment module 21 in the embodiment shown here in a substantially linear movement, one in the x-direction 51 and z direction 52 , ie in the horizontal plane from left to right and from front to rear movable transport unit 6 the transport of the carrier cassette 1 takes over. The slides 2 are usually not the carrier cassette for pretreatment 1 taken.

Upon completion of the pretreatment (s), the carrier cassette becomes 1 at the exit station 4 of the pretreatment module 21 output and the input station 3 of the treatment module 22 to hand over. Here too, a linear transport in the x-direction takes place through the treatment module 22 through to its starting station 4 , For transporting the carrier cassette 1 takes care of a trans port unit 7 , Within the treatment module 22 a single treatment of the dewaxed slides is carried out 2 or the sample (s) thereon, z. B. a tissue section. The slides 2 are doing the carrier cassette 1 , For example, with the aid of a vacuum pipette, not shown here, individually at least partially removed, usually subjected to several staining steps, and after each staining step back into the carrier cassette 1 spent. In this case, the carrier cassette 1 in the feed direction 9 either on different stationary reagent pipetting units designed as pipetting units 10 Moving past, the different reagents on the slides 2 Apply or the carrier cassette 1 stays in the treatment module during their stay 22 stationary and a movable pipetting unit 10 moves by means of a transport unit 8th each with reagents on the carrier cassette 1 to, or the carrier cassette 1 moves linearly to the starting station 4 to and one or more moving pipetting units 10 transport reagents. In the treatment module 22 are corresponding (not shown here) reagent reservoir 30 , z. B. antibody reservoirs provided. After immunostaining the carrier cassette 1 with the colored samples at the exit station 4 output.

The storage of slides within the carrier cassette 1 can be done in different ways. For example, the slides become 2 on jetties from the two side walls 24 . 25 the carrier cassette 1 protrude into the interior, filed. The webs may be continuous or interrupted. Also possible is a storage on only four points, for example, four each lying on a plane protrusions within the carrier cassette 1 ,

2 schematically represents a further embodiment of the invention. In this embodiment is a turntable 11 provided, which is rotatable in a horizontal plane. The turntable has at least one bracket 12 which is designed to be a carrier cassette 1 can record. At the in 2 illustrated bracket 12 it is one on axes 15 (S. 3 ) in the vertical direction movable lifting table, on the bottom of a substantially U-shaped to the outer edge of the turntable 11 open web is applied, which is the basis of a carrier cassette 1 encompasses and so slipping the carrier cassette when moving the turntable 11 prevent. Of course, other types of brackets 12 possible.

In the illustrated embodiment, a carrier cassette 1 from an entrance station 3 , which is designed here like a ramp, with the help of a transport device 13 to the turntable 11 transported and there in one of the brackets 12 added. The turntable 11 and the holder 12 have mutually associated recesses 18 . 19 It's the transport unit 13 facilitate the carrier cassette 1 over the ramp 23 in the holder 12 on the turntable 11 to transport. The turntable 11 with the carrier cassette thereon 1 rotates in a horizontal plane, here counterclockwise, by a predetermined angle x, which may be for example about 90 °, and brings the carrier cassette 1 with the slides contained therein 2 in the engagement area of a carrier manipulator 14 , which may be, for example, a vacuum pipette, gripping tweezers or the like.

The carrier cassette can pass through the holder 12 moved in the vertical direction, so raised and lowered again (s. 3 ). In this way, each becomes a slide 2 placed in a position that it the carrier manipulator 14 allows the slide 2 to capture and out of the carrier cassette 1 out move. In the illustrated embodiment, access to a slide occurs 2 from the back of the carrier cassette 1 out. The carrier cassette 1 is therefore in this embodiment both to the front 16 as well as to the back 17 at least partially open.

As in 3 shown, becomes a single slide 2 from the carrier cassette 1 taken by the carrier manipulator 14 detects the slide on a narrow side and advances in the horizontal direction.

By means of an electronic image processing system 26 becomes the surface of the carrier manipulator 14 from the carrier cassette 1 removed slide 2 with the sample on it 20 detected. The electronic image processing system 26 includes a digital camera and other electronic components, for example, for storing the image taken by the camera, as well as an image analysis unit, with the help of which the captured images can be automatically analyzed. The electronic image processing system 26 can z. B. from a digital camera and a properly equipped personal computer (PC) exist. By means of the electronic image processing system 26 will determine the location of the sample 20 detected on the slide surface and determines the area that the sample 20 on the slide surface occupies. In addition, a sample is automatically added 20 determined comprehensive treatment area to be surrounded by the limiting means.

Alternatively, the treatment area can also be determined manually or semi-automatically. This can be done, for example, that a user, e.g. B. with the help of an appropriately equipped computer, generated by the camera and transmitted to a screen image of the sample 20 traverses with the cursor and thus creates the treatment area. In a semi-automatic method, it may be, for example, that an image generated by the camera and analyzed by the image analysis unit and supplemented by a suggestion for the treatment area is displayed to the user on a screen and the user is given the opportunity to change the automatically proposed boundary area to redefine or confirm.

After the area bounded by limiting means on the slide 2 is determined, limiting means is applied along the edge of this area. This is done by means of a Begrenzungsmitteldosierstation not shown in this figure 43 that are either separate or as part of the treatment station 27 can be trained. The delimiter dosing station 43 can from the electronic image processing system 26 determined data are electronically controlled.

4 shows the in 3 illustrated embodiment of the invention, but here additionally the treatment station 27 and a drain device 28 are shown. For a better overview, only reference numbers not yet introduced are used here. 5 additionally shows schematically a plan view. The treatment station 27 has at least one pipetting unit 29 which is movable in all three dimensions, ie in the x, y and z directions. The drainage device 28 is positioned below the site at which the treatment of the slides 2 takes place, so that, for example, in a washing process, from the drain device 28 Liquid collected and over the drain 31 , z. B. in a collection container, can be derived. On one or possibly several shelves 38 that z. B. can be configured as drawers are reagent reservoir 30 accommodated with appropriate reagents by means of the treatment station 27 be dosed. The footprint (s) 38 and / or reagent reservoir 30 are removable.

6 schematically shows in greater detail a section of the plan view according to 5 , Shown is the turntable 11 on which three mounts 12 are visible. The fourth bracket 12 is located in the illustrated operating state below the carrier manipulator 14 and can not be seen for this reason. The brackets are offset with their directed towards the center of the turntable longitudinal axes each offset by 90 ° on the turntable 11 arranged, to the edge. The holder 12a right in 6 is exemplified here as a kind of bracket, the z. B. made of sheet metal, plastic or the like can be made. The turntable 11 is rotatable both clockwise and anticlockwise. A carrier cassette 1 becomes a holder 12 . 12a transported, the turntable 11 turns about 90 ° about its axis and brings the carrier cassette 1 in the engagement area of the carrier manipulator 14 , This is capable of a single slide 2 in the carrier cassette 1 from the back 17 to be grasped and in the horizontal direction to the front 16 push out. The now exposed slide 2 can now be treated and after the treatment step for incubation in the carrier cassette 1 be withdrawn. Subsequently, the carrier cassette can be raised or lowered and another slide 2 the carrier cassette 1 taken for treatment. After completion of all treatment steps, the carrier cassette 1 by means of the turntable 11 transported further and the starting station 4 fed. As shown in the figure, the slide can also be used during the treatment 2 at the treatment station 27 another carrier-loaded carrier cassette 1 supplied and an already finished carrier cassette 1 be dissipated.

7A shows a side view of the in the 2 - 6 illustrated embodiments in section. 7B shows a plan view of the in 7A ramp shown in a side view 23 , A carrier cassette 1 is in a holder 12 , As in 7B shown is the carrier cassette 1 at the bottom with a slot 32 equipped, in which a transport plunger 35 the transport unit 13 intervenes. The transport tappet 35 is with a motor 33 connected, with the help of the transport plunger 35 can be moved in the vertical direction to allow the transport ram 35 with the slot 32 the carrier cassette 1 reversible engagement. A carrier cassette 1 so can at the entrance station 3 by lifting the transport ram 35 grabbed and over the ramp 23 for mounting 12 on the turntable 11 be transported, where the transport tappet 35 is lowered to the carrier cassette 1 release and the rotational movement of the turntable 11 to enable. The motor 33 is on a transport rail 34 arranged. Along this transport rail 34 the carrier cassette is over the ramp 23 in the holder 12 transported. The recesses 18 . 19 in turntable 11 and bracket 12 serve to the transport ram 35 to allow a free movement.

8th shows a simplified schematic view of another embodiment of the invention. In this embodiment, a pretreatment module 21 and a treatment module 22 intended. Both modules 21 . 22 are arranged side by side and functionally connected to each other the. The treatment module 22 has an entrance station 3 on, the one with slides 2 loaded carrier cassette 1 can record. In the illustrated embodiment, a transport device, not shown, for example, a corresponding rail system transports the carrier cassette 1 to the turntable 11 , The slides 2 , for example, in the edge region with a preferably machine or electronically readable label 48 (S. 10 ), z. As a bar code or the like, are provided, after the carrier cassette 1 on the turntable 11 was spent using the electronic image processing system 26 analyzed. First, if available, are those in the label 48 encoded information. The marking 48 contains, for example in coded form information about how the sample 20 is to be treated, for example, if a pretreatment in pre-treatment module 21 is provided. Optionally, the slides can 2 now in the same or in different carrier cassettes 1 grouped or arranged according to the intended treatment. slides 2 , which are to undergo a pretreatment, for example, in a common separate carrier cassette 1 be sorted. This can be done by means of the carrier manipulator 14 happen.

The device includes two transfer stations 36 , at which a transfer of a carrier cassette 1 from one module to another, here from the treatment module 22 to the pretreatment module 21 , and vice versa, can take place. The transfer stations 36 correspond to combined input / output stations 3 . 4 , The transfer station 36 of the treatment module 22 is a corresponding transfer station 36 of the pretreatment module 21 across from. A carrier cassette 1 , their slides 2 Pretreatment is to be subjected by a transport unit not shown nä forth to the transfer station 36 of the treatment module 22 transported and the transfer station 36 of the pretreatment module 21 to hand over. In the pretreatment module 21 becomes the carrier cassette 1 optionally from a transport unit 37 transported. The carrier cassette 1 is then from a transport unit 6 captured and with the slides 2 in different, preferably heatable, dipping baths 5 incubated. Here are three dipping baths 5 shown. The number of possible dipping baths 5 but is not limited. The dipping baths 5 are preferably closed with a cover or with covers, which are not shown for clarity. After pretreatment, the carrier cassette becomes 1 with the pretreated, z. B. dewaxed, slides 2 back to the transfer station 36 of the pretreatment module 21 transported back, the transfer station 36 of the treatment module 22 pass and from there to the turntable 11 transported and the treatment station 27 fed.

At another, in the right part of the 8th illustrated transfer station 36 of the treatment module 22 can another module, eg. B. for automatic Eindeckelung the preparations or an automatic microscopy system connected. A module can in turn with a second transfer station 36 be equipped. This makes it possible to interconnect different modules.

The device can also have a separate output station not shown here 4 dispose of a carrier cassette 1 with completely treated slides 2 or non-treatable slides 2 (eg if not readable) is returned to the user.

Reagent storage 30 be in this embodiment on a designed as a drawer, a plurality of drawers or the like footprint 38 provided. The footprint 38 is designed so that the device missing reagents can be supplied in a simple manner. Reagents are added with a pipetting unit 29 taken and on slides 2 applied.

In the 8th illustrated embodiment has a Trägermanipulator 14 , which is designed here as a gripping mechanism. The carrier cassette 1 is here, z. B. by means of the turntable 11 or by means of the transport unit, not shown, movable in the vertical axis, so that the Trägermanipulator 14 the slides 2 remove and at the treatment station 27 or can lead past their components. Of course it is also possible, instead of the turntable 11 the treatment station 27 and the carrier manipulator 14 to move in the vertical axis (s. 9 ), z. B. by means of a liftable table 41 ,

The treatment station 27 In this embodiment, a plurality of components, which are structurally partly combined and partly distributed to different units or stations (see also FIGS 9 and 10 ). The electronic image processing system 26 and the carrier manipulator 14 form in the embodiment, which in 8th is shown, for example, each a structural unit. Assembled into a structural unit here are a delimiter dosing station 43 and a washing station 39 , Of course, the individual components can be arranged arbitrarily. It is preferred, however, to arrange them on one or both sides along a line which is of the preferably linear direction of movement of the carrier manipulator 14 is defined.

Another arrangement of the components of the processing station 27 is in the 9 and 10 shown. So here are the wash station 39 and the carrier manipulator 14 realized as each separate structural units. The electronic image processing system 26 and the delimiter dosing station 43 are in contrast structurally summarized. The washing station 39 comprises a detergent dosing unit 44 and a fan 49 ,

The on the slide 2 attached marking 48 as well as the sample 20 be through the electronic image processing system 26 by means of a digital camera 42 detected. The from the label 48 For example, extracted information is used to determine which treatment of the slide 2 or the sample thereon 20 should be subjected. The sample 20 collected image information is used to sample 20 with a hydrophobic boundary layer to edge. The embodiment of the Begrenzungsmitteldosierstation 43 is shown here by way of example and comprises a metering unit 50 in the direction of movement of the slide 2 vertical horizontal axis can be moved. By coordinated movement of the slide 2 and the dosing unit 50 The limiting means may be in any form on the slide 2 be applied. The success of applying, for example, automatically with the help of the digital camera 42 be checked.

The in the 9 and 10 Washing station shown in detail 39 includes a blower 49 to reagents from the slide 2 to remove. The fan 49 is preferably designed stationary. By the movement of the slide 2 is a directed blowing off the reagents possible. The washing station 39 further includes a detergent dosing unit 44 , by means of which a slide 2 Wash with washing buffer or other liquids to remove remaining reagent residue. By a correspondingly designed valve different wash buffer can be supplied. By moving the slide back and forth 2 between detergent dosing unit 44 and blowers 49 Of course, successive washing and blowing cycles are possible. The washing liquid is via a trained as a collecting tray drainage device 28 derived. The sequence 31 the drip tray 28 is preferably provided with a device that allows to separate harmless from harmful waste. In 10 For example, a switchable valve is provided in the drain 31 integrated so that harmful waste can take a different route than non-hazardous waste. During the washing cycles it is provided that the pipetting unit 10 , which is here designed as reagent dosing unit, the next required reagent from a reagent reservoir 30 takes and after completion of the wash cycle on the slide 2 dosed. This can be done, for example, in a pipetting area 47 the treatment station 27 happen, which is shown hatched here. The pipetting unit 10 can be close to the location with the help of a pipette rinsing station 40 getting cleaned. This can for example be done so that the pipetting unit 10 Rinsing fluid in an inner washing container 46 the pipette rinsing station 40 metered, which are in the inner washing 46 ansam melnde and from there into the surrounding outer rinsing container 45 Overflowing rinse liquid at the same time also the outside area of the pipetting unit 10 flushes. From the outer washing container 45 that with a separate expiration or with the expiration 31 can be connected, the flushing liquid is connected to dissipate. Of course, the outer washing can 45 also be omitted and the discharge of the rinse liquid directly over the drain 31 respectively. When returning the slide 2 in the carrier cassette 1 for incubation is preferably by means of the electronic image processing system 26 controls that the reagent is the sample 20 completely and sufficiently covered. After completion of the process, the dyed cut can be made with the aid of the electronic image processing system 26 to be appraised and handed over to a documentation system. For this purpose, it may be advantageous to use a high-resolution camera system.

1

medium cartridge

2

slides

3

entrance station

4

starting station

5

bath

6

transport unit

7

transport unit

8th

transport unit

9

feed direction

10

Reagenziendosiereinheit

11

turntable

12

bracket

13

transport unit

14

carrier manipulator

15

axis

16

front the carrier cassette

17

back the carrier cassette

18

recess

19

recess

20

sample

21

pretreatment module

22

treatment module

23

ramp

24

Side wall

25

Side wall

26

electronic Image processing system

27

treatment station

28

off device

29

pipetting

30

Reagent storage

31

procedure

32

slot

33

engine

34

transport rail

35

transport plunger

36

Transfer station

37

transport unit

38

footprint for reagent reservoir

39

washing station

40

Pipettenspülstation

41

table

42

digital camera

43

Begrenzungsmitteldosierstation

44

Waschmitteldosiereinheit

45

Outer rinse tank

46

inner rinse tank

47

pipetting

48

Labelling

49

fan

50

dosing

51

x-direction

52

z-direction

QUOTES INCLUDE IN THE DESCRIPTION

This list The documents listed by the applicant have been automated generated and is solely for better information recorded by the reader. The list is not part of the German Patent or utility model application. The DPMA takes over no liability for any errors or omissions.

Cited patent literature

• - WO 99/49295 [0005]
• WO 2004/074847 [0005]
• WO 2004/058404 [0005]
• WO 99/04432 [0006, 0008]
• US 2003/0099580 A1 [0007]
• - WO 99/34190 [0007]
• WO 2000/008511 [0010]
• WO 04/001389 [0010]

Cited non-patent literature

• - ISO 8255-2 [0015]

Claims (23)

Process for the treatment of carrier-fixed material, in particular biological material or with biological material interacting material, a plurality of vertically stacked arranged in a first carrier cassette slides is used, on each of which the material is applied, and where a) each slide at least partially the first carrier cassette is removed, b) at least a treatment agent on the material on the at least partially removed slide is applied, c) the Slides after application of the treatment agent stored in the first or a second carrier cassette will, and d) the material on the slide in the first or the second carrier cassette with the treatment agent is incubated, wherein at least one of the steps a) to c) is automated. Method according to claim 1, characterized in that that steps a) and c), particularly preferably steps a), b) and c) are automated. Method according to claims 1 or 2, wherein e) the treatment agent after the incubation, preferably automatically, Will get removed. Method according to claim 3, characterized that steps a) to e) are repeated until the material is completely treated on the slide. Method according to one of the preceding claims, characterized in that required for the treatment of the material or appropriate amount of treatment agent automatically, preferably based on the material taken on the slide Area, is determined. Method according to claim 5, characterized in that that on the slide manually, semi-automatically or automatically a treatment area is delimited, the treatment area the material includes. Method of treating a person on a microscope slide applied material, in particular biological material or with biological material interacting material, characterized that on the slide manually, semi-automatically or automatically delimited a material covering the treatment area and the materials required or appropriate for the treatment of the material Amount of treatment agent automatically based on the material on the slide occupied area, based markings on the slide characterizing this area or based on the area of the treatment area and is dosed. The method of claim 6 or 7, comprising the following Steps: i) the automatic detection of the material and / or of markings on the slide characterizing the area of the material, ii) automatically detecting the on the slide of material used or marked by the markings Area, iii) automatic delineation of a treatment area on the slide around the material occupied by the material and / or area marked by the marks, iv) the automatic determination of what is required to treat the material or appropriate amount of treatment agent, and v) the automatic application of the determined amount of treatment agent within the treatment area on the slide. Method according to one of claims 6 to 8, characterized in that the delimitation of the treatment area on the slide by applying a limiting means on the slide or by drying the slide surface around the marks taken by the material and / or from the markings marked area takes place. Method according to one of claims 6 to 9, characterized in that the treatment area substantially that occupied by the material on the slide Area corresponds. Method according to claim 9 or 10, characterized in that a hydrophobic agent is used as the limiting agent, preferably natural or synthetic oil, Fat, wax or plastic. Method according to one of claims 9 to 11, characterized in that the limiting means or the dried Area the of the material on the slide occupied area partially or completely encloses. Method according to one of claims 6 to 12, characterized in that the determination of the treatment of the Material required or appropriate amount to treatment agent based on the treatment area or based that occupied by the material on the slide or the area marked by the marks becomes. Method according to one of claims 6 to 13, further comprising vi) automatically controlling the successful application of the treating agent within the treatment area on the slide. Method according to one of the preceding claims, characterized in that the material is biological material or material interacting with biological material that is is preferably selected from the group consisting of cells, Tissues, cell extracts, cell lysates, serum, proteins, nucleic acids, Antibodies and lectins. Device for carrying out the method according to one of Claims 1 to 15, characterized by an electronic image processing system (FIG. 2) designed as an automatic material recognition system ( 26 ). Apparatus according to claim 16, characterized by a limiting agent metering station for automatically applying the limiting means or for producing the dried surface on the material-loaded carrier ( 2 ). Apparatus according to claim 17, characterized in that the Begrenzungsmitteldosierstation is connected to the electronic image processing system, that the Begrenzungsmitteldosierstation based on data provided by the electronic image processing system ( 26 ) is controllable. Device according to one of claims 16 to 18, characterized in that the device via an input station ( 3 ) designed and adapted to provide a carrier cartridge ( 1 ), the carrier cassette ( 1 ) is preferably configured and arranged such that the slides ( 2 ) are vertically above one another arrangeable. Device according to one of claims 16 to 19, characterized in that a carrier manipulator ( 14 ), which is designed and set up so that a microscope slide ( 2 ) individually and automatically the carrier cassette ( 1 ) and a treatment station ( 27 ), and the slide ( 2 ) after treatment in the carrier cassette ( 1 ) or a second carrier cassette ( 1 ) can be stored. Device according to claim 20, characterized in that the carrier manipulator ( 14 ) is electronically controlled. Device according to one of claims 20 or 21, characterized in that the carrier manipulator ( 14 ) is a vacuum pipette, gripping tweezers or a robot arm. Device according to one of claims 20 to 22, characterized in that the slide ( 2 ) with the aid of the carrier manipulator ( 14 ) is movable along at least one coordinate axis.