WO2014009067A1 - Dispositif et procédé d'incubation de prélèvements de patients - Google Patents

Dispositif et procédé d'incubation de prélèvements de patients Download PDF

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Publication number
WO2014009067A1
WO2014009067A1 PCT/EP2013/061739 EP2013061739W WO2014009067A1 WO 2014009067 A1 WO2014009067 A1 WO 2014009067A1 EP 2013061739 W EP2013061739 W EP 2013061739W WO 2014009067 A1 WO2014009067 A1 WO 2014009067A1
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WO
WIPO (PCT)
Prior art keywords
holder
substrate carrier
liquid
incubation
samples
Prior art date
Application number
PCT/EP2013/061739
Other languages
German (de)
English (en)
Inventor
Winfried STÖCKER
Norbert Rottmann
Original Assignee
Euroimmun Medizinische Labordiagnostika Ag
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Euroimmun Medizinische Labordiagnostika Ag filed Critical Euroimmun Medizinische Labordiagnostika Ag
Publication of WO2014009067A1 publication Critical patent/WO2014009067A1/fr

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L9/00—Supporting devices; Holding devices
    • B01L9/52—Supports specially adapted for flat sample carriers, e.g. for plates, slides, chips
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F31/00—Mixers with shaking, oscillating, or vibrating mechanisms
    • B01F31/20—Mixing the contents of independent containers, e.g. test tubes
    • B01F31/23—Mixing the contents of independent containers, e.g. test tubes by pivoting the containers about an axis
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0822—Slides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0457—Moving fluids with specific forces or mechanical means specific forces passive flow or gravitation
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/54—Labware with identification means
    • B01L3/545—Labware with identification means for laboratory containers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • G01N1/31—Apparatus therefor
    • G01N1/312—Apparatus therefor for samples mounted on planar substrates

Definitions

  • the invention relates to a device and a method for incubating patient samples, which are intended for immunological and / or histochemical examination and which are in the form of tissue or cells.
  • the patient samples are brought into contact with a liquid containing at least one reactant, thus producing staining patterns which are evaluated as part of a study of the sample.
  • tissue sections or cells are first stained and then the stained structures are examined in the context of a report.
  • tissue sections are produced and assessed on the microscope.
  • Specimens used in histological work include, in particular, surgical specimens, specimen excisions and biopsied tissue, the primary aim of which is to examine such stained tissue sections for the reliable detection and typing of tumors.
  • the processing of tissue sections to a stained specimen involves numerous steps, either manual or at least partially automated.
  • the contacting of the samples with the respectively required liquids, especially reactants and / or washing liquids usually takes place by immersion in the liquid, dripping of the liquid, placing the slides with the cut down on the surface of the liquid or passive penetration or active filling of the Liquid in a capillary gap.
  • test preparations arranged in parallel on a slide in particular tissue sections or nitrocellulose membranes with proteins derived from whole antigen extracts, are immersed, as it were, upside down in a liquid patient sample, in particular blood or serum become.
  • a reagent carrier with grooves, into each of which a liquid is introduced is provided and the test preparations arranged in rows on a slide are immersed for incubation in the respective associated channel.
  • the slide is placed in a pivoting motion together with the reagent carrier.
  • test systems known in the field of histology and histopathology are often that relatively expensive reactants are required for the incubation of the samples in many cases.
  • space required for machines is already considerable in the case of a medium sample volume. Therefore, there is a great need in the aforementioned field of laboratory diagnostics for test systems and devices with which as many different samples as possible can be processed flexibly and at least partially simultaneously. It is particularly important that very good and reliable test results can be achieved despite reduced costs of the examination and smaller space requirements of the devices.
  • the invention is based on providing a device and a method, with the patient samples, especially tissue sections, flexible, rapid and highly accurate to the presence of pathogens, antibodies and of selected biological substances.
  • the contacting of the individual samples with a reactant and / or a washing liquid take place in such a way that on the one hand an intimate contact between the liquids and the sample is produced and on the other hand a sufficient convection in these liquids is ensured.
  • the invention first relates to a device for incubating patient samples intended for immunological and / or histochemical examination, which are in the form of tissue and / or cells.
  • the device has at least one substrate carrier for applying at least one of the patient samples and at least one holder that can be equipped with at least one substrate carrier.
  • the device according to the invention has an incubation unit, by means of which the at least one patient sample arranged on the substrate carrier can be incubated with a liquid.
  • the device embodied according to the invention is characterized in that at least one movement means is provided, by which the liquid is movable relative to the patient sample at least at times during the incubation.
  • Incubation in this context means the period of time following a first contact between liquid and patient sample, in which the sample is brought into contact either continuously or at intervals with one liquid at a time, so that it leads to a reaction, in particular to formation an antigen-antibody complex, between the solid-phase patient sample and at least one liquid reactant.
  • a reaction in particular to formation an antigen-antibody complex
  • the liquid is thoroughly mixed with the reactants therein, and, on the other hand, a particularly intimate incubation of the patient sample is achieved.
  • the generation of a relative movement between the patient sample and the fluid used as needed is carried out according to a first specific embodiment by the substrate carrier, the holder and / or the incubation unit with the aid of a suitable motion is pivoted.
  • the pivoting movement causes a movement of the liquid such that it flows at least partially along the surface of the substrate carrier and wets the patient sample located thereon.
  • the holder is designed as part of the incubation unit, wherein the holder is pivoted together with the substrate carrier to produce the relative movement between sample and substrate carrier.
  • the swiveling device preferably has an electric motor which, either directly or in a preferred embodiment, indirectly initiates the pivoting movement of the at least one substrate carrier via a gear.
  • the incubation unit preferably has a fluid intake, in which the respective fluid can flow in different directions. It is conceivable to carry out the liquid absorption depending on the shape and size of the patient sample or of the substrate carrier as a groove or as a trough.
  • the substrate carrier is for incubation, preferably with the patient sample a surface of the liquid facing, so to speak head first, immersed in the liquid-filled liquid intake. Regardless of how the substrate carrier is aligned relative to the liquid intake, it is also conceivable to insert the substrate carrier either in the liquid receptacle or to attach by at least one fastening means to the receptacle.
  • flow obstacles in particular in the form of elevations, can be provided within the liquid intake, preferably at the bottom.
  • the liquid receptacles are designed such that the liquid therein does not leak during the pivoting movement. This is achieved either by the fact that the grooves or the tub have an additional volume, in the excess medium can flow at least temporarily, or that at least one sealing element is provided on the circumference of the gutter or tub rim.
  • a corresponding seal between the peripheral edge of the trough-shaped or trough-shaped liquid receptacle and the substrate carrier on which the sample to be incubated is preferably carried out.
  • the moving means as a nozzle, blower and / or as a pressure or suction pump, by means of which the liquid in the fluid intake is at least temporarily moved. Furthermore, it is conceivable that with the aid of the pumping unit, the addition of a liquid in the liquid intake or the suction takes place.
  • the technical solution according to the invention can be used particularly advantageously if tissue sections are arranged on the substrate carrier and at least two, preferably a plurality, in particular ten, such substrate carriers are mounted on a holder.
  • the holder has means for releasably securing at least one substrate carrier.
  • the substrate carrier is non-destructively detachable from the holder and can be connected to this at a later date again.
  • a holder in which at least one substrate carrier, preferably a plurality of substrate carriers can be arranged flexibly, enables an optimal, in particular needs-based combination of different samples for their provision, processing, incubation, visual examination and / or archiving.
  • the development of the invention described above is characterized in particular by the fact that, depending on the examination requirement and the time available, different substrate carriers, which in turn can be equipped with individual patient samples in different ways, can be fastened in a holder. It is particularly advantageous that such a holder can be transferred together with the substrate carriers and the samples arranged thereon at least temporarily during the incubation in a pivoting movement.
  • the invention thus provides a flexibly configured device which on the one hand enables the simultaneous incubation of a large number of different samples and on the other hand ensures a particularly effective incubation of patient samples which are in the form of tissue sections or cells.
  • the substrate carriers are arranged within the holder such that their main extension direction extends transversely to the longitudinal axis of the holder. In this context, it makes sense to carry out the substrate carrier significantly smaller than the standard slides used. Such substrate carriers are sometimes referred to as magnetic chips.
  • the substrate carriers preferably have a surface area of 110 to 120 mm 2 , with a preferred substrate carrier having a surface area of approximately 6 x 19 mm 2 .
  • At least two of the described substrate carriers are preferably arranged in a holder, with a plurality of such substrate carriers being arranged side by side in a very preferred manner.
  • On a substrate support is in turn depending on the planned investigation, one or a plurality of samples in the form of tissue or Cells arranged.
  • a patient sample arranged on a substrate carrier is surrounded at least in some areas by a web.
  • the substrate carrier it is likewise conceivable for the substrate carrier to have, as an alternative or in addition, at least one recess within which the sample is located.
  • the web and / or the area surrounding the recess preferably projects beyond the patient sample in such a manner that bleeding of a medium, in particular a mounting medium, from the sample to the surface of the substrate carrier surrounding the sample is reliably prevented.
  • the samples arranged on a substrate carrier can have substantially the same size or, depending on the respective analysis, be of different sizes.
  • the sample is arranged on a carrier substrate and is applied to the substrate carrier together with the carrier substrate.
  • the carrier substrate is a glass or plastic carrier, in particular a fragment of a conventional cover glass or a plastic film.
  • a carrier substrate preferably has a surface area of 10 to 12 mm 2 , wherein a carrier substrate according to a special development of the invention is made square, in particular has a surface of 3.33 x 3.33 mm 2 .
  • a functional coating is provided between the sample and the substrate carrier.
  • a functional coating ensures that the sample, in particular a section of human tissue or a covering glass fragment with a tissue section located thereon, adheres particularly securely to the substrate carrier. It is essential here that no detachment takes place between the sample and the substrate carrier during the incubation.
  • a further preferred embodiment of the invention provides that the substrate carrier and the patient sample are arranged and designed in such a way that at least temporarily a cover can be arranged above the patient sample.
  • a cover is usually a glass cover, for example a cover glass or a part of a cover glass, which is placed over the sample before a visual examination, in particular before a microscopic examination.
  • a mounting medium is located between the cover and the patient-mounted on a substrate support.
  • the mounting medium can be designed as a UV-curable adhesive.
  • the cover rests cleanly and preferably at a defined distance above the sample on a stop surface.
  • the cover is located on a Patient sample at least partially surrounding web or on a peripheral surface of the substrate carrier, which projects beyond the sample on.
  • the patient sample is preferably a tissue section, in particular an at least temporarily paraffinized tissue section.
  • tissue section in particular an at least temporarily paraffinized tissue section.
  • Such cuts are well known in the field of histopathology. As a rule, this is a thin slice of a few micrometers thickness, which originates from a tissue sample of a patient to be examined.
  • the incubation and / or cleaning or washing steps are carried out in the apparatus described above.
  • samples, in particular tissue sections, of the same and / or different size can be arranged on a substrate carrier. Likewise, it is conceivable, depending on the planned examinations or test runs, to arrange substrate carriers variably within the holder.
  • variable means, for example, that a different number of substrate carriers can each be equipped, as required, with one or a plurality of samples originating from different patients and / or having the distinguishing properties, and arranged in a holder.
  • a control is provided according to a specific embodiment of the invention.
  • an identification code in the form of a 2D barcode, a 3D code or an RFID tag is provided on each substrate carrier and / or holder so that each sample can be uniquely identified and localized in a laboratory.
  • an association is generated by means of a laboratory software and / or in the control unit depending on the planned processing of a sample, at least one of the steps to be performed the investigation and / or depending on a property of the investigation to be performed, the at least a patient sample and / or a placement of the holder is based on at least one substrate carrier.
  • the invention further relates to a method for incubating patient samples intended for immunological and / or histochemical examination.
  • the at least one patient sample is provided as a tissue section and / or in the form of cells and applied to a substrate carrier.
  • a holder is provided which is connected directly or indirectly to the substrate carrier.
  • the patient sample is incubated with a liquid while the patient sample is indirectly received in the holder over the substrate carrier.
  • the method according to the invention is characterized in that during the incubation the liquid is at least temporarily stored. is moved relative to the patient sample. Preferably, the liquid is moved so that it flows over the liquid. It is essential that the sample is sufficiently wetted with the fluid and the liquid is constantly moved during the incubation, so that this remains in a mixed state.
  • the at least one substrate carrier is attached non-destructively releasably in the holder and the holder is pivoted together with the substrate carrier.
  • the at least one sample is immersed, at least temporarily, in a liquid receptacle of an incubation unit.
  • the liquid intake of the incubation unit is preferably filled with a reactant and / or a washing liquid, so that the sample is finally brought into contact with the liquid during the liquid movement.
  • the holder, to which at least one substrate carrier can be releasably secured is designed as part of the incubation unit and has at least one fluid intake in the form of a channel or trough.
  • a needs-based placement of the substrate carrier with the individual samples and the holder with the individual substrate carriers takes place.
  • a method according to the invention is characterized in that, depending on the intended processing of the sample, at least one of the method steps of the examination to be performed and / or depending on a property of the examination to be performed, an association is generated which corresponds both to a mounting of the holder with one or more Substrate carriers as well as the substrate carrier with one or more samples is based.
  • the holder can be grasped, positioned and / or moved directly, or that in turn a further device is provided, to which the holder is fastened at least temporarily.
  • additional devices can be either receptacles, in particular casings, or else racks which serve to store and plan to provide the samples arranged inside the holder.
  • a device designed according to the invention and a method based on the essential features of the invention are preferably usable in the field of histology, histopathology and histochemistry, in particular immunohistochemistry. It is essential that a plurality of tissue sections can be flexibly arranged in a holder having a substrate carrier, wherein the arrangement can always be optimized with regard to the method steps to be performed and the time required for this purpose.
  • the witnesses a relative movement between the liquid used in each case and the samples during the incubation allows on the one hand a particularly intensive contact between the liquid and a patient sample and on the other hand ensures a good mixing of the liquid.
  • FIG. 1 substrate carrier with tissue samples arranged thereon;
  • Fig. 3 Holders with populated substrate carriers in different views
  • Fig. 4 representation of different types of equipment of a holder
  • Fig. 5 Top view of a tray with a plurality of holders
  • FIG. 6 shows various views of an incubation unit with channel-shaped liquid receptacles
  • Fig. 7 Schematic representation of the incubation of substrate carriers with tissue samples, which are arranged in trough-shaped fluid receptacles of a reagent carrier;
  • FIG. 8 shows different views of an incubation unit with a trough-shaped fluid receptacle
  • Fig. 10 Detail view of a fastener between the holder and the substrate carrier.
  • FIG. 1 shows a substrate carrier with samples arranged thereon, as it can be used in a preferred manner with the apparatus according to the invention and the corresponding method.
  • the substrate carrier is designed in such a way that it can be applied to or attached to a holder alone or together with others as required, in order to set the substrate carrier together with the holder in a pivoting movement. If no holder is used, it is likewise conceivable to incubate such a substrate carrier individually and / or to feed it to further method steps.
  • a preferred possibility of bringing the samples arranged on the substrate carrier into contact with a liquid, in particular with a liquid which contains at least one reactant or with a washing liquid, is to pass the samples adhering to the substrate carrier quasi upside down into the corresponding fluid immerse.
  • the liquid is in this case in a gutter or tub of an incubation unit, which is pivoted during incubation together with a substrate carrier, which is optionally attached to a holder.
  • the illustrated substrate carrier 1 is characterized in that it is significantly smaller than conventional microscope slides.
  • such substrate carriers 1, which are also referred to below as magnum chips are preferably equipped with more than one sample 2 and / or with more than one biochip, that is to say a carrier substrate 3 coated with biological material, in particular a cover glass fragment.
  • the substrate carriers 1 described can have different sizes, the size always being adapted to the requirements.
  • 1 a shows a plan view and an oblique view of the substrate carrier 1, while FIG.
  • FIG 1 b shows an enlarged detail view of the detail "A"
  • Magnum chip has a size of 6 ⁇ 19 ⁇ 0.6 mm 3.
  • Four samples 2 of biological material, which are paraffinized tissue sections which are applied to cover glass fragments 3, are arranged on the substrate carrier 1 according to FIG Cover glass fragments 3 with the paraffinized tissue sections represent so-called biochips 2 which allow preferential provision, handling and examination of biological material
  • the biochips 2 have a surface area of 3.3 x 3.3 mm 2.
  • the cover glass fragments 3 of the biochips 2 have a thickness of 0.15 mm, while the tissue sections have a thickness of about 0.004 mm, each on the sub stratif 1 biochip 2 represents an independent sample, which can be omitted depending on the examination needs or replaced by another sample and / or another biochip.
  • four biochips 2 are arranged at least approximately in series on the substrate carrier 1, wherein the cover glass fragments 3 with the tissue sections are in turn arranged on the substrate carrier 1 or magnum chip.
  • an identifier 4 in the form of a 2D barcode is provided at one end of the substrate carrier 1.
  • This identifier 4 contains all the information about the sample 2 required for the examination, in particular the type of tissue and the origin of the tissue, in order to identify the samples 2.
  • a corresponding identification code is stored in a central control unit for each sample 2. Both in the assembly of the substrate carrier 1 and in the incubation, examination, diagnosis and archiving of the samples 2, this identification code is taken into account.
  • the control unit is designed in this context such that at least the incubation and the delivery of the samples to an examination unit are automatically controlled.
  • 1 b shows in a greatly enlarged detail view the detail "A" -shown here is a biochip 2 arranged on the substrate carrier 1.
  • Biological material in the form of a tissue section is arranged on the actual chip 3, which is designed as a cover glass fragment b illustrates the size ratio of the thickness of the Substratträ- gers and the cover glass fragment 3 on the one hand to the thickness of the tissue section 5 on the other.
  • devices and devices which are used for the manual or automatic processing of the samples 2 mounted on the substrate carriers 1 or on magnum chips according to FIG. 1 a require significantly less space, energy and reactants if the design is suitable in relation to all steps performed with a sample, in particular incubation, examination, diagnosis and archiving.
  • the theoretically usable surface of a standard slide corresponds approximately to seventeen times the surface of the substrate carrier 1 or magnum chip used.
  • the capacity of four samples 2 on a comparatively small, bar-code carrying substrate carrier 1 is particularly well suited for the typical requirements of a histological or histopathological diagnostic laboratory, eg in tumor diagnostics.
  • a large number of samples 2 of different patients can be dyed rationally, effectively and flexibly;
  • the individual biochips 2 of 3.33 ⁇ 3.33 mm 2 are still large enough to be evaluated by one or two in most questions small number of biochips to ensure optimal diagnosis.
  • FIG. 2a shows a substrate carrier 1 with the dimensions according to FIG. 1, on which not a plurality of samples 2 but rather a sample is arranged.
  • FIG. 2b shows, by way of example, special combinations of differently equipped substrate carriers 1 in a holder 6, wherein the type of a sample, the dimensions of the biochips and / or the size and number of samples per substrate carrier 1 can always be adjusted as required to use the principal advantages of the invention optimally.
  • FIG. 2 a shows a substrate carrier 1 on which a 2D barcode is attached as identification 4 and on which a tissue section is located.
  • the substrate carrier 1 has the dimensions 48 ⁇ 19 ⁇ 0.6 mm 3 .
  • the use of large-area tissue sections in addition to smaller tissue samples is quite useful for certain applications in the field of medical diagnostics.
  • the substrate carrier 1 in turn has suitable fastening means, so that it can be fastened in a holder 6.
  • FIG. 2 b shows an arrangement of various substrate carriers 1, which can be assembled in a preferred manner and fastened in a holder 6.
  • the substrate carriers 1 have either tissue sections of different number and size or an arrangement of biochips 2.
  • the combination of the required substrate carrier 1 with the samples 2 biological material thereon is made as needed and can always meet the requirements of the investigation and / or the work progress, in particular Dependence of the next required work step to be adapted in the laboratory.
  • the substrate carriers have suitable fastening elements, so that either a substrate carrier or a plurality of substrate carriers, for example ten, can be fastened to a holder as required.
  • the holder is connected directly or indirectly with a pivoting device such that the holder 6 with the substrate carriers 1 attached to it, on which the samples are located, can be pivoted about a longitudinal axis.
  • the holder is pivoted 6 as soon as the samples with a liquid, in particular with a liquid which has at least one reactant, or with a washing liquid, have been incubated.
  • the pivoting device has a pivotable tray on which the holder is placed for pivoting, or the pivoting device in turn has fastening elements and / or recordings on which the holder 6 can be releasably attached.
  • a fastening and / or releasing the holder on the pivoting device is destructive and preferably without tools.
  • FIG. 3 a shows a top view of a holder 6 with ten substrate carriers 1 fastened thereto.
  • the holder 6 is designed in the form of a frame to which the substrate carriers 1 are releasably secured by means of suitable fastening means.
  • the substrate carriers 1 are arranged transversely to the longitudinal direction of the holder 6 and have an identifier 4 which is embodied as a 2D code and in each case via four samples 2 of biological material which are arranged on corresponding adhesion surfaces.
  • an identifier 7 is provided in the form of a 2D code, so that the holder 6 with the substrate carriers 1 and samples therein can be identified at any time unambiguously and is thus reliably supplied to the required operation and / or archiving ,
  • a clip mechanism or via an adhesive connection, which is optionally also designed to be easily detachable.
  • FIGS. 3b to 3d show the holder 6 according to FIG. 3a with the substrate carriers 1 fastened thereto in sectional or detailed views.
  • 3b shows a view along the section "A - A" which runs centrally in the longitudinal direction of a substrate carrier 1 and transversely to the longitudinal axis of the holder 6.
  • Four samples 2 are arranged on the substrate carrier 1 in the longitudinal direction of the substrate carrier 1.
  • an identifier 4 is applied at one end, which permits unambiguous identification of the substrate carrier 1 and the samples 2 located thereon.
  • the samples of biological material are tissue sections which are based on the presence of specific tumor cells and / or tumor markers to be examined.
  • FIG. 3c shows the detail "A" in an enlarged detail view, clearly showing the substrate carrier 1 with the biochip 2 arranged thereon, on which the tissue section 5 intended for an examination is located each biochip 2 carrying a tissue section is covered by a cover 8, which is adapted in shape and size to the surface of the holder 6 equipped with substrate carriers 1. Between the cover 8, which rests on its outer periphery in the edge region of the holder 6, and the samples 2 there is a mounting medium, which is needed for the visual examination of the samples 2. Usually, used in either a water soluble or an organic mounting medium. Alternatively, in some cases, an adhesive film coated with an appropriate medium is used.
  • FIG. 3 shows the detail "B" also in an enlarged detail view: While the biochip 2 rests on the substrate carrier 1 with its supporting glass substrate 3, a sample 2 of biological material in the form of a tissue section is located on the surface Tissue section is covered by a cover 8, wherein a mounting medium is provided between cover 8 and biochip 2.
  • the holder 6 provided according to the invention offers the advantage that for the substrate substrate 1 after staining with the samples 2 or biochips thereon again in others Thus, in a holder 6, the substrate carriers 1 can be reassembled with the samples of a patient, which, however, have passed through different staining protocols and thus were assigned to other functional groups during the staining process.
  • the substrate carriers 1 are fixed at least temporarily and detachably, e.g. through the use of clips.
  • the holder 6 is an aluminum frame, on which a cover 8 in the form of a size-matched cover glass is applied.
  • the size of the holder 6 shown corresponds to the size of a conventional normalized slide, so that this holder 6 can be used together with commercially available microscopes, which are aligned on conventional slides.
  • the size of the holder 6 predetermines the size of the substrate carrier 1, in particular how many magnum chips 1 equipped with biochips 2 and / or macrochips with large-area tissue sections can be arranged together.
  • the dimensions are preferably selected such that ten substrate carriers 1 or magnum chips equipped with biochips 2 are arranged in a holder 6.
  • FIG. 6 The format of this holder 6 allows containers for storage of conventional slides to be used for storage of the holder 6, optionally together with or separately from conventional slides.
  • Figures 4a to 4c show in an illustrative form in each case a holder 6, as it is used for the visual examination of the samples by means of a microscope, with substrate carriers 1 mounted therein.
  • the arrangement of the samples 2 on the substrate carriers and the substrate carrier 1 within the holder 6 was in this case made such that a visual evaluation of the samples 2 can be made particularly effective, in particular the required travel paths between the individual samples 2 and the focal plane of optics Examination unit, in particular the lens of a microscope can be minimized.
  • At least one assignment is generated in a controller on the basis of the required examinations and of an analysis plan, according to which the placement of the substrate carriers 1 and the holder 6 takes place.
  • an assignment is created specifically for each working step during the processing and examination of the samples and taken into account in the execution of the individual examination steps.
  • FIG. 4a While in the holder 6 shown in FIG. 4a ten substrate carriers 1 are fixed with samples 2 arranged in the form of biochips, in the holders 6 according to FIGS. 4b and 4c, larger substrate carriers 1 are provided in addition to substrate carriers 1 with biochips 2, have been applied to the large-area tissue sections.
  • the substrate carriers 1 in the holder 6 according to FIG. 4a each have a control 9 and two or three biochips 2 with tissue sections.
  • the arrangement of the tissue sections P1 to P4 on the substrate carriers 1 and with respect to the holder 6 was carried out depending on the patient affiliation and the colorations to be carried out.
  • the assignment of the individual tissue sections to individual discrete examination sites on the substrate carriers 1 and within the holder 6 was generated by means of a control unit.
  • FIG. 5 shows a tray 10, in which in turn a plurality of holders 6 are combined with the substrate carriers 1 fastened therein.
  • a tray 10 may be preferably fed to the microscope stage of a microscope for the visual inspection of the individual samples 2.
  • it is suitable for an advantageous, especially space-saving archiving of the samples 2. Due to the identifiers 4, 7, 1 1, which are provided on the substrate carriers 1, the holders 6 and the tray 10, at any time a unique identification of the individual Samples 2 possible.
  • FIG. 6 shows an incubation unit 12 with which the samples 2 of biological material, in particular tissue sections, fixed to the substrate carriers 1 or magnum chips are effectively and readily reproducible with the required reagents and / or rinsing liquids the liquid phase to bring into contact, so are incubable.
  • the holder 6, which serves to accommodate different substrate carriers 1, represents the central component of the incubation unit 12.
  • Figure 6a shows a holder 6 of the incubation unit 12 in an oblique view
  • Figure 6b provided the same for the incubation Holder 6 in a plan view and a longitudinal "A - A" and a cross section "B - B" of the holder 6 shows.
  • the holder 6 of the incubation unit 12 has a plurality of, in the example shown, five fluid receptacles 13 in the form of grooves.
  • the grooves are in this case arranged parallel in the body of the holder 6 and respectively provided for receiving a stocked with samples 2 substrate carrier 1 and a Magnumchips.
  • individual Magnumchips for incubation with the samples arranged thereon facing upwards or the gutter floor, so to speak, upside down, are inserted.
  • the holder 6 of the incubation unit with the liquid receptacles provided therein is designed in such a way that the holder 6 can be set into a pivotal movement about its longitudinal axis, in order to move the liquid in the channel-shaped liquid receptacles into a flow movement along the grooves offset.
  • the incubation unit 12 which has a holder with five grooves 13 for receiving substrate carriers 1, represents an advantageous embodiment of the invention. In this case, a group of five substrate carriers 1, together with the samples 2 thereon, undergoes a multiplicity of working steps in the laboratory, especially the incubation.
  • the holder 6 of an incubation unit 12 five substrate carriers 1 each having four biochips 2 are arranged so that the holder has a total of 20 independent samples 2. These samples 2 undergo time-synchronous incubation with a descending alcohol series medium, immunohistochemical staining and incubation with an ascending alcohol series medium.
  • the incubation is carried out by first filling the respectively required liquid 14 into the grooves 13 of the holder and then displacing the holder 6 with the substrate carriers arranged in the grooves in a pivoting movement about its longitudinal axis.
  • the liquid 14 reciprocates during the incubation in the individual grooves 13, whereby a good mixing of the liquid 14 is ensured and an intimate contact between the sample 2 and the liquid 14 takes place.
  • the grooves 13 of the holder 6 shown in Figure 6 protrude on both sides beyond the end of the substrate carrier 1, so that in this area an additional reservoir 15 is located in which excess liquid 14 collects before it flows back to the other side.
  • the channel bottom is inclined in the longitudinal and in the transverse direction, so that a slight trough shape is created.
  • the inclination in The filling and / or removal of a liquid 14 into or out of the grooves 13 is usually carried out in the area of the additional reservoirs 15.
  • the longitudinal direction is shown in section "B-B"
  • twenty slides would have to be picked at least 20 times to perform the corresponding work steps, whereas according to the illustrated embodiment, an entire group consisting of 20 samples 2 and one holder 6 would be handled, and liquids would be handled, for example
  • the outlay for the processing of the individual samples is thus considerably reduced.
  • FIG. 6 the incubation of the samples 2 is shown in detail in FIG.
  • the illustrated holder 6 of an incubation unit 12 as well as the substrate carriers 1 correspond to those which have been explained in connection with FIG.
  • the incubation of the samples 2 takes place in that the holder 6 is pivoted with the substrate carriers 12 contained in the channels 13 or trays in such a way that the liquid 14 alternates in the two longitudinal directions of the channels 13 or 13 Tubs moved.
  • the reactions in the tissue sections incubated in this way are remarkably uniform because the reactants in the liquid are constantly mixed.
  • FIG. 8 shows an alternative embodiment of an incubation unit 12.
  • the holder 6 of the incubation unit 12, as a fluid receptacle 14, does not have individual grooves, but the over a comparatively large tub.
  • Substrate carriers 1 are preferably incubated with a holder 6 designed in this way, on which large-area tissue sections are located in comparison to the biochips 2. Incidentally, the incubation is effected here by pivoting the holder 6 of the incubation unit 12.
  • each of the patient samples for each of the requested antibodies would have to be reared at least one slice on each slide:
  • the biopsy of the potential tumor is sufficiently large (larger than 7 ⁇ 7 mm) and the biopsate in the HE overview stain is sufficiently homogeneous, it may be sufficient to use only one incision to obtain a biochip therefrom 2 to produce
  • Figure 9 shows a holder 6 which is attached to a receptacle or placed on a tray and with the aid of the receptacle or the tray about its longitudinal axis 17 is pivotable.
  • the longitudinal axis 17, by which the holder 6 is pivoted at least temporarily in the direction of the arrow during an incubation, is shown in phantom in FIG.
  • Attachment elements 16 are again provided on the frame-shaped holder 6, by means of which a plurality of substrate carriers 1 with the tissue sections arranged thereon can be detachably fastened fixedly to the holder 6. It is possible to arrange only one or more, in the illustrated embodiment, up to eight, substrate carrier 1 side by side and to attach to the holder 6.
  • the substrate carrier 1 are arranged transversely to the pivot axis 17, so that after contacting the sample 2 with a liquid 14, the liquid 14 due to the pivotal movement parallel to the longitudinal axis of the substrate carrier 1, the sample 2 flows over, thus it to a forced relative movement between the fluid 14 and the respective sample 2 comes.
  • the incubation takes place, as shown for example in FIG.
  • the intended for the incubation or washing liquid 14 is within a designed as a trough or gutter liquid receptacle 13 of the holder 6 of an incubation unit 12 and is finally put together with the holder 6 and the substrate carriers 1 attached thereto in a pivoting movement.
  • a suitably suitable holder 6 of an incubation unit 12 with grooves is shown in FIG. 6 and with a trough in FIG.
  • the substrate carriers in each case individually into the grooves of an incubation unit 12, as shown in Figure 6, or the holder 6 according to FIG. 9 for incubation of the samples 2 in the tub of the incubation holder according to FIG. 8, or even releasably attached thereto.
  • the liquid 14 flows back and forth due to the pivoting movement within the trough or the channels of the incubation unit 12, so that on the one hand the liquid 14 is always thoroughly mixed and on the other hand intimate contact between the samples 2 adhering to the substrate carrier 1 and the liquid 14 , in particular a liquid containing reactants, or a washing liquid comes.
  • the substrate carriers 1 can be equipped very flexibly with regard to the number and the design of the samples 2. In the illustrated embodiment, there are either up to five tissue sections drawn on carrier substrates 3, which occupy a square base area, or larger tissue sections on a substrate carrier 1. It is clear that the illustrated arrangement of flexibly stockable substrate carriers 1 in a holder 6 allows a plurality of different mounting variants of the holder 6. Depending on the need for the planned examination, the individual samples 2 are arranged in such a way that in particular the demand for the reactants required for the incubation is minimized. In this connection, the swiveling movement carried out during the incubation ensures that, on the one hand, there is always an intimate contact between the reactants and the samples 2 and, on the other hand, that the reactants used are well mixed.
  • a device for the flexible attachment of substrate carriers 1, which is embodied in the manner of the holder 6 shown in FIG. 9, furthermore offers the advantage that a combination of substrate carriers 1 which can be selected as required can be moved, provided, incubated relatively in their entirety relatively an examination unit can be positioned and / or archived. Nevertheless, the system is characterized by a particular flexibility, since the composition of substrate carriers 1 in a holder 6 can be changed easily and at any time and adapted to changing needs.
  • both the individual substrate carriers 1, the so-called magnum chips, and the holder 6 have an identification 4, 7 in FIG Shape of a barcode, preferably a data matrix code.
  • an identification 4, 7 in FIG Shape of a barcode preferably a data matrix code.
  • the samples 2 can be clearly identified, located and fed to the desired method step at any time with the aid of laboratory software and the information stored in a laboratory control.
  • 9 shows in detail the detail "A" marked in FIG. 9. It can be clearly seen that at least one fastening element 16 is provided by means of which a substrate carrier 1 can be simply and reliably connected to the holder 6.
  • the fastening element 16 has a coupling point to which, for example by means of a latching or clip element, a secure and yet non-destructively releasable connection between the holder 6 and a sample carrier 2 equipped with substrate 2 can be produced
  • the fastening element 16 is designed in such a way that unintentional release of the connection, in particular during the pivoting operation and also after a relatively long time, is reliably precluded thus sure that a u Different number of substrate carriers 1 in a simple manner, quickly, safely and still be arranged with very high flexibility as a whole in a holder 6.

Abstract

L'invention concerne un dispositif et un procédé d'incubation de prélèvements de patients, destinés à un examen immunologique et/ou histochimique et se présentant sous la forme de tissus ou de cellules. Typiquement, notamment dans le domaine du diagnostic tumoral, tous les prélèvements ou des zones partielles choisies de ces derniers, dont l'examen s'avère d'un intérêt particulier, sont tout d'abord disposés individuellement ou en groupes sur des lamelles. Ensuite, une incubation est effectuée avec un liquide contenant au moins un agent réactif, et/ou avec un liquide de lavage. Ainsi les prélèvements de patients sont colorés et les motifs de coloration générés peuvent être exploités dans le cadre d'un examen des prélèvements. La solution technique selon l'invention est caractérisée en ce que, d'une part, les lamelles (1) peuvent être disposées dans le support respectivement individuellement ou conjointement avec d'autres lamelles et, d'autre part, notamment pendant l'incubation, un mouvement relatif se produit entre les prélèvements de patients (2) et le liquide utilisé pour l'incubation. Le mouvement du liquide par rapport au prélèvement garantit tant une incubation intime des prélèvements qu'un bon mélange du liquide.
PCT/EP2013/061739 2012-07-11 2013-06-06 Dispositif et procédé d'incubation de prélèvements de patients WO2014009067A1 (fr)

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DE102012013680.3A DE102012013680A1 (de) 2012-07-11 2012-07-11 Vorrichtung sowie Verfahren zur Inkubation von Patientenproben
DE102012013680.3 2012-07-11

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EP3459636A1 (fr) * 2017-09-22 2019-03-27 Euroimmun Medizinische Labordiagnostika AG Module d'incubation
EP3599013A1 (fr) * 2018-07-25 2020-01-29 Tecan Trading Ag Appareil de mélange
EP3838415A1 (fr) * 2019-12-16 2021-06-23 Universidade de Évora Support de protection pour sections minces, procédés et utilisations associés
US11125735B2 (en) 2017-03-30 2021-09-21 Championx Usa Inc. Dynamic wax deposition testing systems and methods

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DE20317944U1 (de) * 2003-11-18 2004-03-11 Heiser, Volker, Dr. Probenträger für Moleküle
EP1970747A1 (fr) * 2007-03-16 2008-09-17 MetaSystems Hard & Software GmbH Dispositif de maintien pour porte-objet
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Publication number Priority date Publication date Assignee Title
US11125735B2 (en) 2017-03-30 2021-09-21 Championx Usa Inc. Dynamic wax deposition testing systems and methods
EP3459636A1 (fr) * 2017-09-22 2019-03-27 Euroimmun Medizinische Labordiagnostika AG Module d'incubation
WO2019057900A1 (fr) * 2017-09-22 2019-03-28 Euroimmun Medizinische Labordiagnostika Ag Dispositif d'incubation et système équipé d'un dispositif d'incubation et table basculante
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EP3599013A1 (fr) * 2018-07-25 2020-01-29 Tecan Trading Ag Appareil de mélange
EP3932532A1 (fr) * 2018-07-25 2022-01-05 TECAN Trading AG Appareil de mélange, installation de mélange et procédé de mélange de substances dans des récipients fermés
EP3838415A1 (fr) * 2019-12-16 2021-06-23 Universidade de Évora Support de protection pour sections minces, procédés et utilisations associés

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