US20040266015A1 - Automated sample processing apparatus and a method of automated treating of samples and use of such apparatus - Google Patents

Automated sample processing apparatus and a method of automated treating of samples and use of such apparatus Download PDF

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Publication number
US20040266015A1
US20040266015A1 US10/741,628 US74162803A US2004266015A1 US 20040266015 A1 US20040266015 A1 US 20040266015A1 US 74162803 A US74162803 A US 74162803A US 2004266015 A1 US2004266015 A1 US 2004266015A1
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United States
Prior art keywords
reagent
probe
reagents
mixing
cup
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Abandoned
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US10/741,628
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English (en)
Inventor
John Favuzzi
Marc Key
Lars Winther
Ole Rasmussen
Bob Lathrop
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Dako Denmark ApS
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DakoCytomation Denmark AS
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Priority to US10/741,628 priority Critical patent/US20040266015A1/en
Assigned to DAKOCYTOMATION DENMARK A/S reassignment DAKOCYTOMATION DENMARK A/S ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: WINTHER, LARS, RASMUSSEN, OLE F., FAVUZZI, JOHN, KEY, MARC, LATHROP, BOB
Publication of US20040266015A1 publication Critical patent/US20040266015A1/en
Assigned to DAKO DENMARK A/S reassignment DAKO DENMARK A/S CHANGE OF NAME (SEE DOCUMENT FOR DETAILS). Assignors: DAKOCYTOMATION DENMARK A/S
Priority to US11/338,524 priority patent/US7603201B2/en
Priority to US12/494,542 priority patent/US20100017030A1/en
Priority to US13/345,605 priority patent/US9182324B2/en
Abandoned legal-status Critical Current

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    • G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • G—PHYSICS
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Definitions

  • This application relates to the field of sample processing systems and methods of processing samples.
  • the present invention may be directed to the automated processing, treatment, or even staining of samples arranged on carriers, such as slides, and in some embodiments, directed to the continuous or batch processing of samples and carriers, as well as washing elements of a sampling system.
  • Embodiments may further relate to control systems for sample processing and data acquisition, data maintenance, and data retrieval for sample processing.
  • Applications to which the present invention may especially relate include immunohistochemistry, in-situ hybridization, fluorescent in-situ hybridization, special staining, and cytology, as well as potentially other chemical and biological applications.
  • the present invention relates to an automated sample processing apparatus for treating samples arranged on carrier means, such as microscope slides or sample tubes, located at defined positions close to or in the apparatus by aspirating a portion of selected reagent from a station containing a plurality of reagents and thereafter dispense the reagent to a sample, e.g. a tissue, organic cells, bacteria etc., arranged on the carrier means.
  • carrier means such as microscope slides or sample tubes
  • the invention also relates to a method of automated treating of samples by mixing reagents and dispensing the mixture to the sample.
  • a sample is in this application to be understood as a biological sample such as histological samples, e.g. tissue and cell specimens, including cell lines, proteins and synthetic peptides, tissues, cell preparations, blood, bodily fluids, bone marrow, cytology specimens, blood smears, thin-layer preparations, and micro arrays, and specifically biological samples on microscope slides.
  • histological samples e.g. tissue and cell specimens, including cell lines, proteins and synthetic peptides, tissues, cell preparations, blood, bodily fluids, bone marrow, cytology specimens, blood smears, thin-layer preparations, and micro arrays, and specifically biological samples on microscope slides.
  • Sample processing in immunohistochemical (IHC) applications and in other chemical and biological analyses may require one or a number of various processing sequences or protocols as part of an analysis of one or more samples.
  • the sample processing sequences or protocols may be defined by the individual or organization requesting an analysis, such as a pathologist or histologist of a hospital, and may be further defined by the dictates of a particular analysis to be performed.
  • a biological sample may be acquired by known sample acquisition techniques and may comprise, for example in IHC applications, tissues generally or even in some applications one or a plurality of isolated cells, such as in microarray samples, and may be presented on a sample carrier such as a microscope slide. Furthermore, the sample may be presented on the carrier variously and potentially in some form of preservation. As one example, a sample such as a layer or slice of skin may be preserved in formaldehyde and presented on a carrier with one or more paraffin or other chemical layers infiltrating the sample.
  • Immunologic and histological applications may require processing sequences or protocols that comprise steps such as deparaffinization, target retrieval, and staining, especially for in-situ hybridization (ISH) techniques.
  • steps such as deparaffinization, target retrieval, and staining, especially for in-situ hybridization (ISH) techniques.
  • ISH in-situ hybridization
  • Previous efforts to automate sample processing may be deficient in several aspects that prevent more robust automated sample processing, such as: the lack of sufficient computer control and monitoring of sample processing; the lack of information sharing for processing protocol and processing status, especially for individual samples; the lack of diagnostic capabilities; and the lack of real-time or adaptive capabilities for multiple sample batch processing.
  • Sample processing apparatuses for staining and treating samples by means of probes normally comprises a first station for containing one or more reagent vials; a second station for mounting slides, a probe arranged to aspirate a portion of reagent from a selected reagent vial and dispensing the reagent to a slide on which the sample is arranged and a drive means for moving the probe between the various stations.
  • U.S. Pat. No. 5,948,359 discloses an apparatus of the above mentioned type, wherein the first station comprises a vial holder for holding 40 or more vials in order to provide a wide range of different reagents adapted for different staining purposes, and thereby the possibility of automatically staining a large number of slides requiring different staining processes.
  • the apparatus facilitates that many different staining processes can be performed at the same time in the apparatus, because this avoids the necessity of batching samples requiring the same procedure or other treatment with reagents, and processing each batch individually.
  • U.S. Pat. No. 5,723,092 discloses a sample dilution well for an immunoassay analyser in which a sample and a diluent is mixed by rotating the sample dilution well.
  • Samples such as plasma or urine are loaded into the immunoassay analyser in a number of sample containing tubes, and a probe is arranged for collecting a portion of the sample in a selected sample carrying tube and transferring the sample to the dilution well, adding a portion of diluent, such as water to the sample portion and mixing the two by rotating the dilution well, after which a portion of the diluted sample is aspirated from the dilution well for examination.
  • a probe is arranged for collecting a portion of the sample in a selected sample carrying tube and transferring the sample to the dilution well, adding a portion of diluent, such as water to the sample portion and mixing the two by rotating the dilution well, after which a portion of the diluted sample is as
  • the staining procedure is laborious and uses many different reagents.
  • the staining protocol may include the following steps: deparaffination, washing, antigen retrieval, endogenous biotin or enzyme blocking, incubation with immunological reagents, molecular probes, secondary visualization reagents and various chromogen reagents, washing steps and counterstaining.
  • An on-board mixing device should be able to mix a multiple of reagents and mixtures.
  • Non-limiting examples include:
  • Dilution of chromogens concentrates, mixing and dilution of two, three or four component enzyme chromogen reagents, dilution of buffer concentrates, dilution of immunological reagents with dilution buffer, dilution of visualization reagents with dilution buffers, mixing several visualization reagents with dilution buffer or dilution of enzyme blocking reagents, dilution of biotin blocking reagents or mixing and dilution of counterstaining reagents.
  • the chromogen reagents e.g. DAB, AEC, fast red etc
  • DAB dibenzoic acid
  • AEC adenosine triphosphate
  • fast red adenosine triphosphate
  • Chromogens like the Fast Red alkaline phosphatase chromogen are made ready for use by mixing and dilution of two or three reagents, which are very different in nature with regard to salt content, viscosity and density. Furthermore, the resulting mixtures are unstable over time and need to be used within a short time.
  • Some chromogens suitable for e.g. horseradish peroxidase, like DAB and AEC, are easily oxidized when exposed to air during e.g. vigorous mixing or dilution.
  • the enzyme chromogens and counterstain reagents like e.g. hematoxylin, are semi oxidized and can contain precipitates and solids. By further oxidation or slight change in pH, the reagents can further precipitate.
  • Antibody and enzyme containing reagents often contain stabilizing proteins and or detergents, which causes the solution to foam when being shaken or stirred. Many proteins cannot easily tolerate to be exposed to the hydrophobic air in foam.
  • Wash buffers can contain detergents, which can foam when shaken or stirred. The foam can spread to other compartments of the instrument in an unwanted and unpredictable way.
  • Spill over/carry over must be avoided.
  • the staining process is characterized by using many, complex and very different reagents and buffers and in many different dilution ratios and mixtures. Some of the reagents or buffers are incompatible with each other's. In the event of cross contamination due to e.g. carry over, the reagents may be ruined within seconds or solids can precipitate, making the staining unsuccessful.
  • enzyme containing reagents can not be mixed with the corresponding chromogens, or high salt concentrates may not be mixed with e.g.
  • proteins containing mixtures, or organic solvents can not be mixed with protein containing mixtures, or highly pH buffered wash buffers can not be mixed with low buffered mixtures without significantly altering the properties of the reagents. Accordingly the cleansing and washing of the mixing device need to be very efficient.
  • the mixing device should ideally be able to mix very different volumes of reagents in both small and large volumes ratios.
  • the degree of dilution and mixing ratios of reagents may vary from small to high ratios.
  • the mixing device should ideally allow:
  • the object of the present invention is to improve the known apparatuses for sample processing as well as the method for automatic sample processing by facilitating a wider range of available processes of treatment, so as to increase the number of different staining and/or treatment processes that may be performed automatically, alternatively or additionally to provide an increased quality of some specific staining processes.
  • staining or treatment processes requiring a mixture of unmixable reagents such as a water based and an oil based reagent, or insoluble reagents
  • a mixture of unmixable reagents such as a water based and an oil based reagent, or insoluble reagents
  • the quality of the staining process may be improved as a desired degree of mixing of reagents may be provided or an optimal application time window for a deteriorating mixture may be reached.
  • sample processing apparatus comprising a reagent mixer having
  • a mixing cup for receiving or more broadly adapted to receive two or more reagents and mixing means for mixing or more broadly a mix element adapted to mix the reagents in the mixing cup, and
  • An apparatus may comprise an automated sample processing system comprising a plurality of drawers, a plurality of sample carrier retainment assemblies each removably configured with one of the drawers, and an adaptive sample processing control system to which the drawers and the sample carrier retainment assemblies are responsive.
  • the sample carrier retainment assemblies may comprise slide retainment assembly and may be removably configured with the drawers to provide sample processing with the drawers of the system.
  • the adaptive sample processing control system may automate the sample processing system such that one or more batches of samples may be processed according to one or more protocols, potentially indicated by information on the slides that may be automatically identified by the adaptive sample processing control system.
  • Sample processing may comprise one or more sampling protocols and steps, such as deparaffinization, target retrieval, and staining.
  • staining is used for the end product of the process, by which certain parts of the sample may be stained, i.e. have obtain a different colour, either in the optic range or in another electromagnetic range, such as ultra violet, or the staining may be an detectable, preferably automatically detectable, change in properties, such as fluorescent properties, magnetic properties, electrical properties or radioactive properties.
  • the sample normally have to undergo a series of treatment steps, such as—but not limited to—washing, binding of reagents to the specific parts of the sample, activation of the reagents, etc. and each treatment step may include a plurality of individual treatments.
  • reagents prepared from two or more separate reagents which may be somewhat incompatible e.g. unmixable, such as a water based and an oil based reagent, or insoluble, and therefore requires that the two or more reagents are manually prepared and introduced into a reagent vial shortly before starting the staining process in order to obtain the best possible staining result for the selected examination purposes.
  • different staining process steps require a mixture of the same two reagents but in different dissolution ratios.
  • Some process step may require a mixture of two or more reagents that, when mixed, becomes unstable, e.g.
  • the present invention relates to a sample processing apparatus for treating samples arranged on carrier means or carrier, comprising
  • a vial station for containing at least two reagent vials, a carrier means station or more broadly a carrier station arranged for intermediate storage of a plurality of carrier means or carriers,
  • probe drive means or more broadly a probe drive arranged for moving or more broadly adapted to move a probe
  • the probe drive means is arranged to aspirate a portion of reagent from a selected reagent vial of the vial station by means of or even with a probe and to apply reagent to a selected carrier means of the carrier means station,
  • the apparatus comprises a mixing station with a reagent mixer having a mixing cup for receiving or adapted to receive two or more reagents and mixing
  • [0055] means for mixing or more broadly a mixer element adapted to mix the reagents in the mixing cup, and
  • the vial station is a collection of a plurality of vials, at least two, but often 20-60 vials or more, which may or may not be physically arranged in close proximity to each other.
  • the term station does not indicate that the vials must be located within one, confined area; rather it indicates the existence of a plurality of vials.
  • the probe drive means or even probe drive may be a robot arm with two or three degrees of freedom, such as an articulated arm or one track or a set of perpendicular tracks along which a probe retainer of the probe drive means may be displaced, wherein the probe retainer may be moved in a direction normal to the track or tracks.
  • the skilled person may readily design other types of probe drive means, e.g. combinations of the above described.
  • the carrier means or even carrier may be provided to the apparatus in a two-dimensional array, e.g. constituted by individual rows of carrier means as discussed in the example below, or the carrier means may be provided in any manner known in the art, e.g. arranged in a carrousel or as a row of carrier means.
  • the carrier means may also be arranged movably with respect to the probe drive means, such as in an endless row that is advanced automatically past the operating area of the probe drive means or as a two-dimensional array that may be moved in a direction perpendicular to a travel direction of the probe drive means, so that the probe may reach any carrier means by the combined movement of the probe and the array.
  • the carrier means are preferably arranged in groups or series on trays or the like, so that a plurality of carrier means may be removed from or inserted into the apparatus simultaneously, and the apparatus preferably also comprises means for performing the intermediate storage of the carrier means with samples thereon and the removal of the carrier means from the apparatus automatically.
  • control means typically a computer having a central processing unit and one or more memory unit associated therewith, means for controlling or more broadly a control element the various operations of the apparatus by controlling step motors, solenoids, valves and/or other drive or control parts of the apparatus.
  • the control means may have one or more data communication ports for enabling data communication with external computers by wire or wireless.
  • the control means does not have to be physically arranged within the apparatus itself but may be a computer external to the sample processing apparatus and connected to the apparatus via a data transmission port thereof.
  • the probe drive means is arranged to apply the reagent mixture from the mixing cup to selected carrier means.
  • the mixed reagents from the mixing cup may be applied to the samples by separate means, such as a separate probe and probe drive means.
  • the carrier with the sample in question may be moved to a location at an outlet from the mixing cup.
  • the probe drive means for the task.
  • the mixing cup may receive the two or more reagents to be mixed from a separate set of vials or other reagent sources, but it is for rationalisation of the apparatus preferable that at least some of the reagents to be mixed may come from the same vials as are used for containing reagents to be applied to the samples.
  • the probe drive means is therefore arranged to aspirate portions of reagents from at least two selected reagent vials of the vial station and apply said portions of reagents to the mixing cup.
  • the mixing means or more broady a mix element of the reagent mixer may advantageously be constituted by cup drive means or more broadly a cup drive arranged for cyclic movement of the mixing cup, e.g. shaking or rotation in a horizontal or a vertical plane, so as to mix reagents contained in the mixing cup.
  • the mixing of the reagent may be further improved by arranging mixing elements, such as blades or edges within the cup and stationary with respect to the cup.
  • Other known mixer types may instead be preferred, such as shaft-driven impellers or magnetically driven impellers.
  • the cyclic movement is preferably a rotation of the mixing cup, advantageously about a substantially vertical axis.
  • the rotation is in a preferred embodiment an intermittent rotation in a clockwise and in an anticlockwise direction.
  • the rotation speed may be varied through control means acting upon a driving motor in such manner that the fluid content inside the mixing cup is agitated, but at so low speed that substantially all reagents remain in the mixer cup.
  • the speed may be controlled depending on the type of reagent in the mixer cup.
  • the speed is controlled according to the processing protocol controlling the processing of the sample(s) to be processed in the apparatus.
  • the rotation pattern is controlled by controlling means or more broadly a control element, such as a computer, according to the sample processing protocol, thereby optimising the quality the mixing in the mixer cup in respect to the reagents present in the mixer cup.
  • a control element such as a computer
  • sample processing protocol is meant a sequence of processing steps defined for the actual sample treated in the sample processing apparatus.
  • the mixing cup has inner walls extending upwardly and outwardly from a bottom part of the mixing cup, and the cup drive means is capable of rotating the mixing cup at an angular speed sufficient to fling all reagents contained therein out of the mixing cup.
  • the inner walls are smooth and may terminate in an upper rim at the widest inner diameter of the mixing cup, in which case the high-speed rotation, preferably about a symmetry axis of the cup, will cause the waste reagent or cleansing liquid to be flung out over this upper rim.
  • the inner walls of the cup extend upwards and inwardly above the level of the widest inner diameter, at which one or more exit openings are provided in the inner wall as outlets for the waste reagent or the cleansing liquid.
  • the top may be open for the probe to enter the mixing cup, or it may be closed to facilitate that the cyclic movement of the mixing cup for mixing the reagents may have a vertical component.
  • the probe may in this case enter the mixing cup through one of the exit openings.
  • the reagent mixer comprises a waste reagent collecting chamber having a sidewall part laterally surrounding the mixing cup and arranged to collect reagents flung out of the mixing cup by rotation of the cup.
  • the reagent mixer further comprises a mixing cup holder for receiving and supporting the mixing cup in a releasable manner, so that different mixing cups may be used for different reagent mixtures.
  • replacing means are provided for replacing the probe that is moved by the probe drive means with another probe, so as to avoid contamination of the reagents in the various reagent vials by the use of more than one probe.
  • This may be combined with a probe washing station, in which one probe is washed after it has been replaced at the probe drive means so that it is clean and ready for repeated use without risking contamination of the content of the reagent vials.
  • the present invention also relates to a method of fully automated treating of samples arranged on carrier means by means of a sample processing apparatus controlled by means of a control means, wherein the method comprises the steps of:
  • the steps of dispensing portions of the first and the second reagents comprises the steps of
  • the method further comprises the step of replacing the probe arranged with the probe drive means with another probe between handling two different reagents or between handling a reagent and a reagent mixture.
  • the present invention further relates to the use of an apparatus of the present invention as described above for exercising the method of the present invention. More specifically the invention relates to the use of the apparatus for IHC and ISH and sample processing of the type using special stains as well as batch-mode sample processing.
  • Support should be understood to exist for the following aspect and embodiment of the invention: use of an apparatus according to any of the apparatus claims for exercising the method according to any of the method claims for immunohistochemistry and in-situ hybridization. Also support should be understood to exist for use of an apparatus according any of the apparatus claims for exercising the method according to any of the method for batch-mode sample processing.
  • FIG. 1 is a perspective view of a sample processing apparatus according to the invention
  • FIG. 2 is a plan view of a sample processing apparatus according to the invention.
  • FIG. 3 is a perspective view of a detail of the sample processing apparatus according to FIG. 2,
  • FIG. 4 is a perspective view of a reagent mixer according to the invention.
  • FIG. 5 is a vertical cross-section of the reagent mixer according to FIG. 4.
  • the present invention includes a variety of aspects, which may be combined in different ways.
  • the following descriptions are provided to list elements and describe some of the embodiments of the present invention. These elements are listed with initial embodiments, however it should be understood that they may be combined in any manner and in any number to create additional embodiments.
  • the variously described examples and preferred embodiments should not be construed to limit the present invention to only the explicitly described systems, techniques, and applications. Further, this description should further be understood to support and encompass descriptions and claims of all the various embodiments, systems, techniques, methods, devices, and applications with any number of the disclosed elements, with each element alone, and also with any and all various permutations and combinations of all elements in this or any subsequent application.
  • FIG. 1 shows one schematic embodiment of a sample processing system 1 in accordance with the present invention.
  • Cabinet sections form outer portions of the system and serve to address general structural considerations of the system (a top cabinet section is not shown in FIG. 1).
  • the sample processing system may comprise a plurality of drawers used for the handling and processing of samples and sample carriers such as slides, potentially microscope slides. Other sample carriers may be accommodated consistent with the present invention.
  • Each drawer may be configured to accommodate sample carrier retainment assemblies, such as slide retainment assemblies, carrier racks, modules, or magazines.
  • a sample carrier retainment assembly may comprise a slide retainment assembly.
  • the slide retainment assembly may comprise a slide rack, module, or magazines.
  • Slide retainment assembly is configured to accommodate a plurality of slides in at least one configuration in corresponding sample carrier retention devices.
  • the sample carrier retainment assemblies are utilized in the processing of samples as further described below. It should be further noted that the sample carrier retainment assembly can be removably configured with the drawers, and may be stackable or nested within other retainment assemblies.
  • One or more drawers may be provided to accommodate processing materials such as reagent containers 3 for sample processing, also further described below.
  • a processing material retainment assembly such as a container rack may be utilized to accommodate reagent containers or other processing materials within each of drawers 6 .
  • Bottle inserts may be preferably configured with the retainment assembly to ensure proper processing material positioning within the processing material retainment assembly and the drawer.
  • Multiple drawers allow for one or a plurality of sample processing protocols to be performed by the system 1 .
  • Past efforts at sample processing may have been limited to processing sequences for an entire batch of carriers within the system.
  • the present invention may allow for multiple batch processing, including real-time or adaptive capabilities for multiple batch processing, as further described below.
  • the system may access, use and wash multiple probes or syringes for pipetting or otherwise aspirating fluids required for the staining of samples mounted or otherwise presented on slides.
  • a system with a single reusable probe may wash the probe between each fluid applied.
  • the task of washing the probe can have a large impact on the throughput of the overall system.
  • the present invention may allow for multiple probes to be available to the system for use.
  • the system may continuously have a clean, uncontaminated, sterilized, or an unused probe available to use and sample processing is not impacted by the required cleaning routine.
  • the cleaning routine may be necessary to eliminate the possible cross contamination of fluids and, in some embodiments, may take up to about 1 minute to accomplish.
  • the cumulative impact of the cleaning routine on a series of processing steps can add time to the throughput capabilities of the system.
  • the addition of multiple probes or syringes may eliminate this impact and significantly decreases the time required to process the samples.
  • Embodiments of the present invention comprise a mixing station or reagent mixer 9 (best shown in FIGS. 4 and 5).
  • the system may mix component fluids, such as dyes, buffers, or other processing materials, preferably on demand and as the processing steps and protocols dictate. Fluids required during the processing steps may sometimes need to be mixed with other fluids to create a final activated fluid. However, the activity levels of these mixtures can be time sensitive and may therefore only be effective for a short period of time.
  • the on demand mixing of fluids is advantageous in that it allows the fluids to be mixed immediately before being used.
  • the syringe or probe 10 in preferred embodiments, will aspirate fluids into and from the mixing station 9 to mix component fluids. A rinse may further be dispensed into the mixing station to sterilize the station.
  • FIGS. 1 and 2 a presently preferred embodiment of a sample processing apparatus, also called a staining apparatus 1 , according to the invention is shown in FIGS. 1 and 2.
  • the sample processing apparatus 1 comprises a rectangular frame 4 surrounding a first station 2 comprising an array of compartments wherein each compartment a reagent vial 3 is placed, and a second station 5 wherein a number of separate racks 6 (which may be a drawer) is placed, and where each rack comprises a number of microscope slides 7 mounted side by side in the rack 6 .
  • each rack may hold up to 8 slides, but the rack may be designed to hold any suitable number of slides. With eight racks arranged side by side, the shown embodiments may hold up to 64 slides 7 each having a sample, e.g. a tissue mounted on the upper side of the slide, so that reagent may be applied from above to the sample on each slide.
  • a robot arm 20 in FIG. 1 for moving a probe 10 in X and Y (as well as Z) direction as indicated by the arrows X and Y is arranged above the frame 4 of the sample processing apparatus.
  • the robot arm may is therefore position the probe 10 above all reagent vials 3 as well as above all the microscope slides 7 , and may further operate the probe 10 to aspirate portions of reagent contained in any of the vials 3 , to transfer the portion of reagent and apply it to any of the slides 7 in order to provide a selected staining or treatment of the sample on each slide 7 .
  • suitable control means e.g. a computer (not shown) having the appropriate software and input data for the purpose, this staining apparatus I is able to automatically staining or treating samples requiring different staining or treatment reagents and processes.
  • FIG. 2 The same staining apparatus viewed from below the robot arm is shown in FIG. 2, disclosing the probe 10 being manipulated by the robot arm.
  • the probe 10 is raised to an upper position (in a Z direction where it is clear of the vials 3 underneath the probe 10 , but the robot comprises means (not shown) for lowering the probe 10 in order to dip the probe tip 211 into the content of a selected reagent vial 3 and to aspirate a selected amount of reagent for the selected staining or treatment process.
  • the staining apparatus 1 of the present embodiment further comprises at least one probe washing station 8 and a reagent mixer 9 , and the robot arm is furthermore arranged to transfer the probe to the washing station 8 as well as to the reagent mixer 9 .
  • the reagent mixer 9 will be described in detail below with reference to FIGS. 4 and 5.
  • the apparatus comprises a reagent mixer 9 having a mixing cup 213 wherein two or more selected reagents may be placed by means of the robot arm and the probe 10 .
  • the reagent mixer 9 thereby provides on-board mixing of any reagents contained in the reagent vials 3 , and thereby more staining processes, e.g. staining requiring the use of mixing of insoluble reagents, or reagents which may only be effective a short time after mixing, are facilitated to be performed automatically within the staining apparatus without the requirement of human interaction.
  • the mixer 9 comprises a mixing cup 213 for receiving reagents released from the probe 10 or 110 .
  • the mixing cup 213 is placed into a holder 215 by means of a complementary snap fitting means 216 and 217 arranged on the inside of the holder 215 and the outside of the mixing cup 213 , respectively.
  • a motor 218 is arranged for rotating the holder 215 and thereby the mixing cup 213 , either intermittently clockwise and anticlockwise in order to provide a mixing of reagents contained in the mixing cup, or by spinning the holder 215 and thereby the mixing cup 213 in order to fling out waste reagents or cleansing liquid from the mixing cup 213 .
  • the mixing cup 213 is preferable provide with sidewalls 220 extending upwardly and outwardly from the bottom 219 , e.g. forming a frusto-conical cavity, and the mixing cup 213 has an upper rim 214 allowing the reagents to escape from the mixing cup 213 during the spinning process.
  • the reagent mixer 9 furthermore comprises a housing 221 having sidewalls 222 surrounding at least the rim 214 of the mixing cup 213 and thereby forming splash faces for collecting any liquid flung out from the mixing cup 213 .
  • the housing also comprises a lid 223 for enclosing a space 224 surrounding the mixing cup 213 in order to avoid reagent spills outside the space 224 .
  • the lid 223 has a central opening allowing reagents from the probe 10 to be dispensed into the mixing cup 213 from above the reagent mixer 9 as well as allowing the probe 10 to enter the mixing cup 213 for collecting the mixed reagents.
  • the housing also comprises a hose connection 227 for draining waste reagent or cleansing liquid from the space 224 , and a tap 226 is arranged for dispensing cleansing liquid into the mixing cup 213 when required.
  • a releasable connection 212 is provided between the probe 10 and the robot arm, enabling the replacement of the probe 10 held by the robot arm by placing the probe 10 in one of a number of free washing stations 8 , where it is released by the releasable connection 212 , and where a new probe 10 ′ is connected to the robot arm by means of the releasable connection 212 .
  • the control means of the apparatus operates the robot arm to commence a staining or treatment run by firstly moving the probe to a first reagent vial 3 , into which the probe tip 211 is inserted into the liquid and is aspirated into the probe 10 in an amount corresponding to the number of samples to be stained or treated, in accordance with the input data provided to the control means.
  • the probe 10 is subsequently, in a first operating mode moved by the robot arm towards the slide rack system 5 in which the slides 7 are mounted.
  • the slides 7 are situated with the surface horizontally oriented and the probe 10 releases the required amount of reagent on the appropriate slides in accordance with the input data.
  • the probe 10 is in a second operating mode moved by the robot arm towards the reagent mixer 9 where it releases the reagent into the cup 213 of the reagent mixer 9 , and is subsequently moved to the probe washing station 8 , where the probe 10 is either washed or—in the alternative embodiment—released into a free washing station 8 , and another probe situated in another washing station is connected to the robot arm.
  • the robot arm moves the new clean probe to a second selected reagent vial 3 for collecting a selected amount of reagent from the second vial 3 , and the probe is thereafter by means of the robot arm moved to the reagent mixer 9 , where the reagent in the probe is dispensed into the cup 13 of the mixer containing the first selected reagent.
  • the second operating mode can according to the invention be commenced several times if more than two reagents are to be mixed for a specific staining or treatment process.
  • the reagent mixer 9 mixes the reagents in the cup 213 thereof, and a clean probe 10 picked up from the washing station 8 by the robot arm is lowered into the cup 213 of the reagent mixer 9 to collect the mixed reagents, where after the robot arm moves the probe 10 towards the second station 5 containing the slides 7 , at which the probe 10 releases the required amount of mixed reagent on selected slides 7 in accordance with the input data.
  • the robot arm with probe 10 is subsequently directed to a free washing station 8 , and the probe 10 is either washed or alternatively replaced by a clean probe, where after the process in accordance with the first or the second operating mode may be repeated or continued with a new reagent or reagent mixture.
  • the present invention is highly suitable for on-board mixing in an automated cytological/IHC/ISH instrument.
  • the shape of the cup allows debris, fines, precipitates or other solids of both high and low density to be centrifuged up and out of the cup at high rotation speeds.
  • the speed may be controlled by control means operating according to the processing protocol carried out for the samples present in the apparatus, i.e. if the reagents present in the mixer cup have high viscosity it may be advantageous applying a very high speed of rotation to ensure that all reagents are removed from the cup.
  • the shape of the cup allow for the specific separation of immiscible liquid phases, by first removing the liquid with the highest density during centrifugation. It should be understood that the shape of the cup makes the mixing almost independent of the volume. The mixing action is the same for a wide range of volumes in the same cup. In contrast, the mixing action of e.g. a propeller mixer is depending on e.g. the volume of reagents, propeller size and container size.
  • the rotating action of the cup allows air to escape from the mixture and prevent foaming.
  • the rotating action can suppress the formation and build-up of foam by forcing the air/liquid foam down to the liquid surface.
  • the rotation pattern (speed and direction) is controlled by controlling means, such as a computer, according to the sample processing protocol, thereby optimising the quality the mixing in the mixer cup in respect to the reagents present in the mixer cup, e.g. in such manner that the formation and build up of foam is suppressed.
  • the cup can be covered with a neutral gas like the noble gasses, nitrogen, and carbon dioxide or similar to further protect the reagents during mixing.
  • a neutral gas like the noble gasses, nitrogen, and carbon dioxide or similar to further protect the reagents during mixing.
  • the protective gas is heavier than atmospheric gas. The layer of protective gas can easily be added and removed, and can cover any of the volumes the cup can hold.
  • the reagents mixing may be done in a batch-mode to make mixtures for treating multiple samples.
  • the on board mixing allow for large dilution ratios and the possibility to dilute or mix reagents, using only a small portion of the mixture for treatment of samples, and discarding the rest of the mixture. This is advantageous, as some dilutions ratios are very large, and too small volumes of the concentrated reagent cannot easily be accurately measured and added. Often a minimum volume of reagent to be measured is defined.
  • the on board cup mixing design allow for a very high degree of flexibility in the staining protocol.
  • the basic concepts of the present invention may be embodied in a variety of ways. It involves both mixing techniques as well as devices to accomplish the appropriate mixer element.
  • the mixing techniques are disclosed as part of the results shown to be achieved by the various devices described and as steps which are inherent to utilization. They are simply the natural result of utilizing the devices as intended and described.
  • some devices are disclosed, it should be understood that these not only accomplish certain methods but also can be varied in a number of ways. Importantly, as to all of the foregoing, all of these facets should be understood to be encompassed by this disclosure.
  • each of the various elements of the invention and claims may also be achieved in a variety of manners.
  • the term “element” is to be understood as encompassing individual as well as plural structures that may or may not be physically connected. This disclosure should be understood to encompass each such variation, be it a variation of an embodiment of any apparatus embodiment, a method or process embodiment, or even merely a variation of any element of these. Particularly, it should be understood that as the disclosure relates to elements of the invention, the words for each element may be expressed by equivalent apparatus terms or method terms—even if only the function or result is the same. Such equivalent, broader, or even more generic terms should be considered to be encompassed in the description of each element or action.
  • any claims set forth at any time are hereby incorporated by reference as part of this description of the invention, and the applicant expressly reserves the right to use all of or a portion of such incorporated content of such claims as additional description to support any of or all of the claims or any element or component thereof, and the applicant further expressly reserves the right to move any portion of or all of the incorporated content of such claims or any element or component thereof from the description into the claims or vice-versa as necessary to define the matter for which protection is sought by this application or by any subsequent continuation, division, or continuation-in-part application thereof, or to obtain any benefit of, reduction in fees pursuant to, or to comply with the patent laws, rules, or regulations of any country or treaty, and such content incorporated by reference shall survive during the entire pendency of this application including any subsequent continuation, division, or continuation-in-part application thereof or any reissue or extension thereon.
US10/741,628 2002-12-20 2003-12-19 Automated sample processing apparatus and a method of automated treating of samples and use of such apparatus Abandoned US20040266015A1 (en)

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US10/741,628 US20040266015A1 (en) 2002-12-20 2003-12-19 Automated sample processing apparatus and a method of automated treating of samples and use of such apparatus
US11/338,524 US7603201B2 (en) 2003-12-08 2006-01-23 Systems and methods for the automated pre-treatment and processing of biological samples
US12/494,542 US20100017030A1 (en) 2002-12-20 2009-06-30 Systems and methods for the automated pre-treatment and processing of biological samples
US13/345,605 US9182324B2 (en) 2002-12-20 2012-01-06 Systems and methods for the automated pre-treatment and processing of biological samples

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US10/539,035 Abandoned US20060063265A1 (en) 2002-12-20 2003-12-19 Advance programmed sample processing system and methods of biological slide processing
US10/741,628 Abandoned US20040266015A1 (en) 2002-12-20 2003-12-19 Automated sample processing apparatus and a method of automated treating of samples and use of such apparatus
US10/539,308 Active 2028-03-30 US8216512B2 (en) 2002-12-20 2003-12-19 Apparatus for automated processing biological samples
US10/538,964 Active 2027-05-21 US8257968B2 (en) 2002-12-20 2003-12-19 Method and apparatus for automatic staining of tissue samples
US10/538,745 Expired - Lifetime US7400983B2 (en) 2002-12-20 2003-12-19 Information notification sample processing system and methods of biological slide processing
US10/539,561 Active 2027-04-06 US7960178B2 (en) 2002-12-20 2003-12-19 Enhanced scheduling sample processing system and methods of biological slide processing
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US10/539,192 Active 2029-02-04 US9778273B2 (en) 2002-12-20 2003-12-22 Isolated communication sample processing system and methods of biological slide processing
US12/076,516 Active 2024-06-01 US7937228B2 (en) 2002-12-20 2008-03-19 Information notification sample processing system and methods of biological slide processing
US12/630,665 Expired - Lifetime US7758809B2 (en) 2002-12-20 2009-12-03 Method and system for pretreatment of tissue slides
US13/073,345 Expired - Fee Related US8386195B2 (en) 2002-12-20 2011-03-28 Information notification sample processing system and methods of biological slide processing
US13/102,181 Expired - Fee Related US8394635B2 (en) 2002-12-20 2011-05-06 Enhanced scheduling sample processing system and methods of biological slide processing
US13/493,969 Expired - Lifetime US8529836B2 (en) 2002-12-20 2012-06-11 Apparatus for automated processing biological samples
US13/567,308 Expired - Lifetime US8663978B2 (en) 2002-12-20 2012-08-06 Method and apparatus for automatic staining of tissue samples
US13/625,115 Expired - Lifetime US9040284B2 (en) 2002-12-20 2012-09-24 Systems and methods of sample processing and temperature control
US13/747,949 Expired - Lifetime US8788217B2 (en) 2002-12-20 2013-01-23 Information notification sample processing system and methods of biological slide processing
US13/758,810 Expired - Lifetime US8673642B2 (en) 2002-12-20 2013-02-04 Enhanced scheduling sample processing system and methods of biological slide processing
US13/964,562 Expired - Lifetime US8784735B2 (en) 2002-12-20 2013-08-12 Apparatus for automated processing biological samples
US14/053,977 Abandoned US20140038232A1 (en) 2002-12-20 2013-10-15 Enhanced Scheduling Sample Processing System and Methods of Biological Slide Processing
US14/151,130 Expired - Lifetime US9599630B2 (en) 2002-12-20 2014-01-09 Method and apparatus for automatic staining of tissue samples
US14/182,616 Expired - Lifetime US8969086B2 (en) 2002-12-20 2014-02-18 Enhanced scheduling sample processing system and methods of biological slide processing
US14/299,178 Abandoned US20140286838A1 (en) 2002-12-20 2014-06-09 Apparatus for automated processing biological samples
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CN116465709A (zh) * 2023-04-19 2023-07-21 杭州海世嘉生物科技有限公司 一种全自动免疫组化染色机控制方法、系统及存储介质

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US9040284B2 (en) 2015-05-26
US20060085140A1 (en) 2006-04-20
EP1573312A4 (fr) 2011-10-12
US20050064535A1 (en) 2005-03-24
US8673642B2 (en) 2014-03-18
US8788217B2 (en) 2014-07-22
US7400983B2 (en) 2008-07-15
EP1573311A2 (fr) 2005-09-14
US7648678B2 (en) 2010-01-19
US8257968B2 (en) 2012-09-04
US20100081167A1 (en) 2010-04-01
US20140234170A1 (en) 2014-08-21
US7758809B2 (en) 2010-07-20
CA2508323A1 (fr) 2004-07-08
US20140186218A1 (en) 2014-07-03
US9778273B2 (en) 2017-10-03
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AU2003287895B2 (en) 2009-07-23
AU2003303390B2 (en) 2009-07-23
EP1573408A2 (fr) 2005-09-14
CN100424166C (zh) 2008-10-08
CA2508113A1 (fr) 2004-07-15
AU2003299740A1 (en) 2004-07-22
AU2003301103B2 (en) 2010-06-10
CA2508113C (fr) 2017-10-31
US20140286838A1 (en) 2014-09-25
CA2507960C (fr) 2017-02-07
AU2003297369A1 (en) 2004-07-22
WO2004058404A3 (fr) 2004-09-10
US20110269238A1 (en) 2011-11-03
US8216512B2 (en) 2012-07-10
WO2004058404A2 (fr) 2004-07-15
US20140356935A1 (en) 2014-12-04
AU2008202863A1 (en) 2008-07-24
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US20060088928A1 (en) 2006-04-27
US8529836B2 (en) 2013-09-10
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US20130029409A1 (en) 2013-01-31
US20060063265A1 (en) 2006-03-23
US8969086B2 (en) 2015-03-03
EP1579194A4 (fr) 2011-12-28
US20060172426A1 (en) 2006-08-03
US8386195B2 (en) 2013-02-26
AU2003300291B2 (en) 2010-09-09
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EP1572979A1 (fr) 2005-09-14
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US8663978B2 (en) 2014-03-04
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US8298815B2 (en) 2012-10-30
US20060088940A1 (en) 2006-04-27
US20130084567A1 (en) 2013-04-04
US20130217108A1 (en) 2013-08-22
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US8394635B2 (en) 2013-03-12
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WO2004059441A2 (fr) 2004-07-15
AU2003299740B2 (en) 2011-04-21
CA2974221A1 (fr) 2004-07-15
CA2508070A1 (fr) 2004-07-15
AU2003303444A1 (en) 2004-07-22
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CA2507960A1 (fr) 2004-07-15
US20060046298A1 (en) 2006-03-02
US8784735B2 (en) 2014-07-22
WO2004059287A3 (fr) 2005-01-27
US20130330252A1 (en) 2013-12-12
WO2004059297A1 (fr) 2004-07-15
US10156580B2 (en) 2018-12-18
AU2003287895A1 (en) 2004-07-14
US9229016B2 (en) 2016-01-05
US7960178B2 (en) 2011-06-14
US20110167930A1 (en) 2011-07-14
AU2003299798A1 (en) 2004-07-22
EP1579190A4 (fr) 2011-12-28
WO2004059288A3 (fr) 2005-01-27
US7937228B2 (en) 2011-05-03
US20080241876A1 (en) 2008-10-02
EP1573408A4 (fr) 2011-10-12
US9599630B2 (en) 2017-03-21
CA2511032A1 (fr) 2004-07-15
CA2508323C (fr) 2017-05-30
EP1576376A4 (fr) 2011-12-28
CA2508070C (fr) 2019-01-22
CN1726386A (zh) 2006-01-25
US20120310422A1 (en) 2012-12-06
EP1579194A2 (fr) 2005-09-28
EP1579190A1 (fr) 2005-09-28
CA2508001C (fr) 2019-03-19
WO2004058950A1 (fr) 2004-07-15
US20130203103A1 (en) 2013-08-08
WO2004059287A2 (fr) 2004-07-15
US20140038232A1 (en) 2014-02-06
EP1573311A4 (fr) 2011-12-28
WO2004059441A3 (fr) 2005-01-20
EP1572979A4 (fr) 2011-10-19
EP1573312A2 (fr) 2005-09-14
US20160084862A1 (en) 2016-03-24
AU2003303390B8 (en) 2009-08-06
US20060045806A1 (en) 2006-03-02
CN100472197C (zh) 2009-03-25
WO2004059288A2 (fr) 2004-07-15

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