WO1999049295A1 - Appareil automatise de coloration - Google Patents

Appareil automatise de coloration Download PDF

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Publication number
WO1999049295A1
WO1999049295A1 PCT/US1998/005919 US9805919W WO9949295A1 WO 1999049295 A1 WO1999049295 A1 WO 1999049295A1 US 9805919 W US9805919 W US 9805919W WO 9949295 A1 WO9949295 A1 WO 9949295A1
Authority
WO
WIPO (PCT)
Prior art keywords
tip
reagent
head
slide
microscope slide
Prior art date
Application number
PCT/US1998/005919
Other languages
English (en)
Inventor
Krishan L. Kalra
Jason Z. Zhang
Zhi-Weng Chang
Jianghong Shui
Original Assignee
Biogenex Laboratories
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 Biogenex Laboratories filed Critical Biogenex Laboratories
Priority to AU69417/98A priority Critical patent/AU762808B2/en
Priority to US09/646,695 priority patent/US6495106B1/en
Priority to PCT/US1998/005919 priority patent/WO1999049295A1/fr
Priority to CA002325583A priority patent/CA2325583A1/fr
Priority to EP98915168A priority patent/EP1066502A1/fr
Priority to JP2000538216A priority patent/JP2002507738A/ja
Publication of WO1999049295A1 publication Critical patent/WO1999049295A1/fr

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Classifications

    • 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
    • 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
    • G01N2001/317—Apparatus therefor spraying liquids onto surfaces
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N35/1081—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices characterised by the means for relatively moving the transfer device and the containers in an horizontal plane
    • G01N35/109—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices characterised by the means for relatively moving the transfer device and the containers in an horizontal plane with two horizontal degrees of freedom

Definitions

  • the present invention relates to automated equipment used in the staining of cells and tissues on microscope slides.
  • U.S. Patent No. 4,985,206 describes an apparatus and process for automating the application of staining reagents to a thin tissue section mounted on a microscope slide.
  • the apparatus and method use a channel-defining element that is assembled with the microscope slide to provide an enclosure of capillary dimensions into which liquids can be injected. Liquids are added sequentially to the capillary space, where the addition of a new liquid forces out the previous liquid.
  • a plurality of these assemblies of microscope slides and specialized covers can be placed in a rack on an apparatus for automated addition of liquids.
  • a further automated immunostaining apparatus known as the Ventana 320TM is produced by Ventana Medical Systems, Inc.
  • This apparatus applies a liquid known as Liquid CoverslipTM to each slide prior to reagent addition.
  • Liquid CoverslipTM is a non- aqueous material having a density less than that of water. When a reagent dissolved in water is added to a microscope slide, the reagent sinks to the bottom of the Liquid
  • CoverslipTM layer spreading across the surface of the slide. Slides are organized on a carousel which rotates beneath a dispensing head of the apparatus for application of reagents or wash fluids.
  • Yet another apparatus known as the Jung Histostainer IgTM Automated Immunostainer, is produced by Leica Instrument GmbH. This is also a carousel-type device, but reagents are applied by a spraying operation rather than by dropping liquid onto an organic film.
  • the apparatus contains a permanent reagent spraying head that can be moved along a single axis to provide spray coverage over a microscope slide located on the rotating tray when the slide is rotated into position underneath the head. Excess reagent is removed by a permanent clearing nozzle which blows air in a pressure front across the slide, forcing excess liquid off at the completion of the reagent incubation step.
  • a further apparatus is the subject of U.S. Patent No. 5,439,649.
  • This device includes an arm moveable in three dimensions attached to a framework.
  • a hollow tip head is carried on the arm, and includes a wash/blow head for dispensing reagents and clearing the slides.
  • the reagent application tip can be attached to the hollow tip head or removed by a pre-selected movement of the arm.
  • an automated staining apparatus comprising a supporting framework, at least one arm moveable in three dimensions attached to the framework; a means for moving the arm, and at least one hollow tip head located on the arm.
  • the hollow tip head will include at least one reagent tip head having a means for releasably engaging a pipette tip by a preselected movement of the arm and a means for alternatively supplying positive or negative gas pressure to the reagent tip head to withdraw or dispense volumes of liquid onto a microscope slide.
  • the hollow tip head will also include at least one wash tip having a means for selectively dispensing a plurality of liquids onto the microscope slide and at least one blow tip having a means for selectively dispensing a gas onto the microscope slide via an exit slit substantially equal in length to the width of the slide.
  • the apparatus will also include at least one pipette tip holder at a predetermined location on the framework adapted for holding a plurality of pipette tips adapted to be releasably attached to the reagent tip head, at least one reagent vial holder at a second predetermined location on the framework adapted for holding a plurality of reagent vials, and at least one microscope slide holder at a third predetermined location on the framework adapted to releasably contain the microscope slide.
  • the apparatus will also include a control means operatively connected to the means for moving the arm, the means for alternatively supplying positive or negative gas pressure, the means for selectively dispensing a plurality of liquids, the means for selectively dispensing gas and the means for controlling movement of said arm between said predetermined locations.
  • the control means will be adapted to cause the tip head to engage a pipette tip, to release the pipette tip, to withdraw a reagent from the reagent vial, to dispense the reagent onto the slide via the pipette tip, and to dispense a gas or a liquid onto the slide through the wash tip and the blow tip.
  • Figure 1 is a top plan view of an embodiment of an apparatus of the invention (shown without a cover on the apparatus);
  • Figure 2 is a front horizontal elevation of an embodiment as depicted generally in Figure 1, showing seven separate representative locations (by thick dashed lines A,
  • Figure 3 is a front horizontal elevation of an embodiment as depicted generally in Figure 2, showing an optional embodiment in which A reagent tip head and optional O-ring are moistened in an optional sponge cup;
  • Figure 4 is a front horizontal elevation of an embodiment as depicted generally in Figure 2, showing the movable arm at position 'F ⁇ wherein the wash tip and blow tip are lowered to proximate the upper surface of a slide for performing wash and blow processing steps;
  • Figure 5 is a front horizontal elevation of an embodiment as depicted generally in Figure 2, showing the movable arm at position 'D', wherein the reagent tip head is lowered to engage a pipette tip;
  • Figure 6 is a front horizontal elevation of an embodiment as depicted generally in Figure 2, showing the movable arm at position 'E', wherein the reagent tip head with a pipette tip engaged is moved to a reagent vial, and the tip lowered to the reagent vial to load the reagent;
  • Figure 7 is a front horizontal elevation of an embodiment as depicted generally in Figure 2, showing the movable arm at position 'G ⁇ wherein the Z head is positioned proximate the microscope slide and the reagent tip head dispenses reagent solution to the slide through the pipette tip;
  • Figure 8 is a front horizontal elevation of an embodiment as depicted generally in Figure 2, showing the movable arm at position 'C ⁇ wherein the Z head has fully retracted and ejected a pipette tip;
  • Figure 9 depicts multiple views including front horizontal elevation, side horizontal elevation, and top plan view of the movable arm and Z head of an embodiment as depicted generally in Figure 1; -6-
  • Figure 10 depicts an embodiment as depicted generally in Figure 9, and in detail depicts a cross section of a portion of the reagent tip head of the Z head;
  • Figure 11 depicts multiple detailed views including elevations and cross sections of the wash tip and the blow tip portions of the Z head;
  • Figure 12 is a top plan view of a number of microscope slides showing the specimen location and bar-code regions of the slide as well as exemplary reagent dispensing patterns for the method of the invention for applying reagents to slides;
  • Figure 13 depicts multiple views including elevations, plan views and perspective views of an individual microscope slide tray used in an embodiment as depicted generally in Figure 1;
  • Figure 14 depicts multiple views including a front horizontal elevation and a top plan view of the slide tray holder, tray and the associated locking structure;
  • Figure 15 depicts a top plan view and side elevation in partial cross section of a reagent vial holder of the invention
  • Figure 16 is a schematic view showing major components of the apparatus that supply gas and liquid to the Z head, along with an indication of an embodiment for the attachment of the supply conduits and other parts associated with the Z head and movable arm;
  • Figure 17 is a top plan view of one trail that the bar-code scanner could trace to read the bar-code on reagent vials and microscope slides. The location indicator codes are also depicted; and
  • Figure 18 depicts multiple views including a side horizontal elevation and a top plan view of an alternative embodiment of the pipette tip holder and the reagent vial holder of the present invention. -7-
  • the present invention provides an automated apparatus for staining cell and tissue specimens on microscope slides, as well as various subsystems for use in the present apparatus and a variety of alternative devices.
  • the apparatus will first be described generally along with its operation, after which the apparatus and various component parts will be described in detail with reference to the figures that form a part of this specification.
  • the apparatus of the invention comprises a supporting framework to which an arm movable in three dimensions is attached. Motors or other means for moving the arm are provided under the control of a computer or other electronic control device that allows programming of movement of the arm between various work locations on or within the framework.
  • a hollow tip head is located on the arm so that liquids or gasses can be dispensed or withdrawn through the head to provide for the various work operations described below.
  • the arm is configured so as to have either multiple, permanently attached tips with different functions or multiple disposable tips located on the arm at the same time.
  • a single hollow tip head is provided having multiple channels connected to separate pumps to which individual tips having different functions will be attached.
  • a portion of the hollow tip head is adapted to pick up disposable plastic pipette tips from the standard containers in which such tips are supplied (for example, Catalog No. 3510-R from E&K Scientific Products, Saratoga, CA).
  • These disposable pipette tips are currently sold in a rack which presents the base of the tip for insertion of a hand-held pipette body into the hollow tip, the tips being arranged in an array so that all individual tips in the container are accessible to the user.
  • the same or similar standard racks of pipette tips can be used in the apparatus of the present invention.
  • One feature of the apparatus of the present invention is the integration of a blow tip and a wash tip in the hollow tip head (hereafter termed the "Z head" because it is the primary component of the present apparatus which moves in the 'Z' dimension).
  • the blow tip there will be an exit slit that is usually substantially equal in length to the width of a microscope slide. If the slit is not equal to the width of a microscope slide with which it is intended to be used, it is preferred that the slit be slightly wider than the microscope slide. A narrower slit is less efficient in removing liquid from the microscope slide surface in the manner as described below.
  • the practical width of the slit is limited by the desire to have a number of microscope slides arranged together in close proximity in the apparatus of the invention and further to avoid wasting buffer or other wash solutions that are applied to a slide through the tip.
  • the exit slit on the blow tip provides a "wall" of gas, typically air, that pushes excess liquid from the surface of a microscope slide as the tip is passed over and parallel to the slide (described below in greater detail).
  • the wash tip is a separate orifice located in close proximity to the blow tip, and is used to deliver diverse liquid solutions to the slide.
  • the framework of the apparatus is also provided with holders at predetermined locations for reagent application tips (hereafter termed pipette tips), among other removable items.
  • reagent application tips hereafter termed pipette tips
  • a holder for a reagent container more typically a plurality of reagent vials (each reagent vial containing, for example, a stain or any of various solutions associated with staining) and a microscope slide holder are also present on the framework at other predetermined locations.
  • standardized motions of the arm can be programmed into the control unit so that individual microscope slides at specific predetermined locations in the microscope slide holder can be treated with reagents and/or wash fluids obtained from reagent vials or from liquids supplied through the hollow tip head on the movable arm.
  • the attachment steps (and optionally the detachment step) for attaching pipette tips to the Z head on the moving arm can be carried out by a pre- -9- selected movement of the arm, much in the same manner that disposable pipette tips are now pressed onto and later removed from the end of a hand-operated pipette.
  • multiple slides each typically having a tissue specimen at some location on its upper surface, are placed horizontally in a tray that is inserted into the apparatus at a predetermined location, usually at a location having registration pins that fit into registration holes in the tray or similar registration means) so that the individual microscope slides are always located in the predetermined relative positions on the frame of the apparatus.
  • the apparatus is programmed as appropriate for the individual slides being treated and reagent vials are placed at their own predetermined locations in the apparatus in the same manner as the tray described above.
  • pipette tips are also made available for engagement and use by the moveable arm. For example, a standard rack of lmL pipette tips can be placed at its predetermined location in the apparatus.
  • the apparatus carries out all preparation, reagent application, incubation, heating (if necessary or appropriate), and specimen rinsing steps to perform the desired staining operation.
  • the Z head on the movable arm moves to each of the slides being treated in a particular cycle and begins by applying the liquid from the wash buffer reservoir via a liquid supply conduit to the wash tip of the hollow tip head.
  • the apparatus will then use the blow tip to remove excess buffer from the slide prior to reagent delivery. This removal is accomplished by blowing gas through the blow tip while the head travels along the length of the slide; a 'wall' of gas exits the slit and removes excess buffer from the slide, without disrupting the tissue specimen.
  • a small amount of buffer desirably remains on the slide to assist in reagent distribution.
  • the Z head on the movable arm then engages a disposable pipette tip from the pipette tip rack that has been inserted into the pipette tip holder in the apparatus.
  • the Z head with the pipette tip attached then takes up a reagent to be applied to a slide or group of slides from a reagent vial in the reagent container holder. For efficiency, a number of slides can be treated with a single reagent at the same time.
  • the reagent is dispensed on the slide in a pre-assigned pattern that operates in combination with the -10- thin liquid film on the microscope slide to assure spreading of the reagent over the entire surface of the slide to which the tissue specimen is attached.
  • the thin liquid film allows less reagent to be used than would be required if the film were not present to assist reagent distribution.
  • the disposable pipette tip is then discarded, and the movable arm moves the
  • the slides are rinsed when the movable arm moves the wash and blow tips to the microscope slide again, and buffer is applied to the slide to rinse off the majority of the reagent.
  • the blow tip then removes the excess buffer from the slide, and the slide is rinsed a second time with the on-line buffer, if desired.
  • This procedure of rinsing and removing excess buffer from a slide is repeated as desired, depending upon the individual stain and the appropriate procedure for rinsing the reagent.
  • the control mechanism generally a programmable computer, keeps track of the time of the various incubations and repeats the steps above as appropriate in order to apply the appropriate reagent to all of the slides that have been inserted into the tray.
  • One feature of the apparatus of the invention that allows efficient use of reagents is the method of spreading reagents described above and further described in detail below.
  • the reagent tends to stay in the location where placed, rather than spreading over the entire surface area of the slide. Since the location of the tissue specimen on a slide is variable, and may not be located in the same place from one slide to the next, automated procedures previously required that the reagent be applied over the entire surface area of the slide.
  • the slide to which a stain reagent will be added is first washed with an aqueous wash solution, usually a buffer, that contains one or more surfactants which reduce the surface tension of water.
  • an aqueous wash solution usually a buffer
  • surfactants which reduce the surface tension of water.
  • it is not deemed entirely satisfactory merely to flood a slide with an aqueous solution of surfactant, since a concentrated reagent added to the slide will then be diluted on the slide.
  • the blow tip of the invention is designed so that excess buffer can be removed from the washed slides to produce a thin film of the aqueous solution.
  • the height of the blow tip exit slit above the microscope slide, the pressure of the compressed gas being blown through the tip, and the rate of movement of the tip are selected to allow a controlled amount of buffer to remain on the slide.
  • the reagents will be diluted as discussed above and will not work optimally. If too little buffer remains, the buffer may evaporate prior to reagent application, and the reagents will not spread sufficiently. Specific techniques for controlling the parameters of the wash tip and blow tip operation to select the desired amount of buffer are described below.
  • the invention also provides for dispensing of reagents on the slide in a pattern that assists spreading.
  • a pattern is selected so that a reagent is not required to diffuse for great distances through the surface film; for example, convoluted application patterns can be selected so that the reagent need not diffuse more than one-fourth (or some other fraction) of the width of the microscope slide.
  • the combination of the buffer film and the application pattern (typically dropwise or in a stream) ensures adequate coverage of the slide regardless of the location of the tissue and allows less reagent to be used than would be required in the absence of a surface film. In a typical -12- operation, the amount of reagent added is less than that which would be required to cover the slide if no aqueous film were present on the slide.
  • the apparatus of the invention can contain additional subsystems for convenience, such as drain pans, reagent vials and other components that are described below in more detail.
  • the apparatus provides two control formats to control the staining process parameters: 'Open' format and 'closed' format.
  • the 'open' format provides great flexibility for the user of the apparatus.
  • the system allows the user to create, change, and adjust numerous system settings, running parameters and staining protocols in the processing of individual tissue specimens on microscope slides to meet different requirements.
  • the 'closed' format the system maximizes the process automation and very little user input is needed. It is especially useful for those users who utilize large batch quantity and similar processing procedures.
  • a bar-code technology can be used in this format.
  • each reagent vial there will be a label affixed at a predetermined location when it is shipped.
  • Three bar-codes are printed in close proximity to each other on the label.
  • the information content of the bar-code can include: 1) Name of the reagent solution; 2) Manufacture date; 3) Expiration date; 4) Serial number; 5) Reagent volume.
  • a human-readable string is also printed on the label and is shown on the side wall of the reagent vial.
  • the content of this bar-code will include the name of the protocol to be used in processing the particular slide.
  • a human-readable string can also be printed immediately under this bar-code as well.
  • the height of the bar-code is generally approximately 0.25 inch, so that there will remain sufficient space for the user to write any other desired information.
  • the coding used in this application will desirably be codel28, as this code can provide advantageous information density. -13-
  • the apparatus moves a laser bar-code scanner around the bar-codes located both on the microscope slides and the reagent vials.
  • the digital computer will then recognize the reagents and the slides to be treated, and will calculate the required volumes of the reagents.
  • a reagent map can also be printed by the computer for user reference. If any deficiency occurs in the reagent supply, the computer will halt the processing and request additional reagent supplies. At the same time, if any unknown protocol is detected, the computer will request that the user create a new one. After this verification, the computer will control the apparatus as discussed above to automatically process the staining operations.
  • Figure 1 depicts an embodiment of the invention in plan view from above.
  • the apparatus of Figure 1 is illustrated without a cover such as would normally form the upper surface of the apparatus and act in concert with retaining walls to enclose the working parts and microscope slides.
  • the front of apparatus 10 is at the bottom of the figure.
  • movable arm 30, which will carry out numerous operations of the apparatus, is visible in its home position in the rear-left corner of the interior of framework 20 which forms the cabinet surrounding the working parts of apparatus 10 (upper-left portion of Figure 1).
  • Framework 20 is formed from various components, such as baseplate 22 and side plate 24, that form the cabinet.
  • the various locations and the corresponding parts of the apparatus or materials that are inserted into the apparatus at these locations are generally visible on baseplate 22.
  • Beneath and slightly in front of the movable arm home position is a square shaped tip disposal orifice 26.
  • a tip disposal bin 28, used for holding discarded pipette tips 90 (described below), is located under the baseplate 22 and is desirably designed as a drawer so that it can be withdrawn from the front of the apparatus 10 (see Figure 2).
  • a horizontal bar 25 is -14- located at the center of the tip disposal orifice 26 and is used to prevent discarded pipette tips from stacking up and blocking the disposal orifice.
  • a drain bin 27 which can simply be a container provided with a drain line to a waste container.
  • holder 100 is adapted to retain in position two standard pipette tip racks 92a and 92b each containing arrays of disposable pipette tips 90.
  • holder 100 for pipette tips 90 are raised regions on baseplate 22 around which the bases of pipette tip racks 92a and 92b fit snugly.
  • reagent vial holder 120 is To the right of pipette tip racks 92a and 92b, in this embodiment in the form of a reagent vial rack.
  • the reagent vial holder 120 can either be affixed to the baseplate 22 or, in the manner described above, it can be adapted to be removable from the baseplate for loading with reagent vials 110 in a more convenient location.
  • the reagent vial holder 120 is adapted to be retained by the baseplate in a predetermined location and orientation, so that any given reagent vial 110 will always be in the same relative position on baseplate 22.
  • each tray 190 is retained in a predetermined location and orientation relative to baseplate 22 and the remainder of the framework 20, so that each microscope slide 130 retained in a well is in a predetermined location relative to the baseplate 22.
  • Movable arm 30 is transported to different locations over baseplate 22 by the action of various motors that operate in combination with sliding tracks to precisely position the movable arm 30 at its desired location within framework 20, in order to carry out the operations described herein.
  • Figure 1 at the top of the figure
  • -15- is the X-axis track 32, in this embodiment the X-axis being the principal longer horizontal axis of the apparatus.
  • a single X-axis track 32 is supported at either end on bearing shafts and brackets 34a (left) and 34b (right).
  • the Y-axis is the principal shorter horizontal axis of the embodiment as shown. Stepping motors are used in these embodiments under the control of the computer or other control apparatus (as described below).
  • a portion of one motor mount 33 for the X-axis motion motor is visible in this figure.
  • the Z-axis in this embodiment is the orthogonal vertical axis perpendicular to the plane of Figure 1.
  • flexible electronic leads and tubing (both gas and liquid supply conduits) would be shown in this figure leading from movable arm 30 to appropriate fluid reservoirs or electronic control equipment. These leads and conduits are not shown in Figure 1 for the sake of clarity, but are described later with respect to specific portions of the apparatus.
  • the supply conduits are desirably sufficiently long and flexible so as to withstand the rigors of frequent use. They will originate from different pumps and desirably will be bound together.
  • the various supply conduits pass through a flexible wire carrier 35 to the left side of X-axis track 32, then through another wire carrier 36 which is at the top of the X-axis track to conduct all supply conduits to movable arm 30 and Z head 70.
  • FIG. 1 The embodiment 10 of Figure 1 is also shown in Figure 2.
  • movable arm 30 In this front horizontal elevation of the embodiment of the invention, movable arm 30 is shown in its home position, to which the arm returns when not otherwise engaged.
  • the home position is desirably selected to minimize interference with other operations, such as the insertion of microscope slides or disposable pipette tips into the cabinet-like interior of framework 20.
  • Figure 2 includes seven thick dashed lines (A, B, C, D, E, F & G) along with the X-axis indicating seven different positions for movable arm 30 (the home position plus six representative operational positions).
  • a 'P' arrow line on movable arm 30 points to the dashed lines to indicates the current locations in the figures.
  • the following table lists the positions and the functions implemented at each position, including moistening the optional O-ring: -16-
  • movable arm 30 is shown at a location where the reagent tip head 40 located on Z head 70 is inside an optional sponge cup 300 suspended from optional sponge cup support 301.
  • an optional O-ring 43 in the reagent tip head 40 can be useful to retain and seal a pipette tip or other device onto reagent tip head 40. If such an O-ring 43 is employed, it can be difficult to pick up and retain pipette tips if the O-ring 43 is dry.
  • the Z head 70 on movable arm 30 can be instructed to move to a position above any interfering part of the apparatus and then move to the position where the reagent tip head 40 is directly above the sponge cup 300 (position B). Then the reagent tip head 40 moves downwardly and inserts into the sponge cup 300, and is submerged into the sponge.
  • the sponge is supplied with water so that the reagent tip head 40 and O-ring 43 will be moistened.
  • the water supply in the sponge cup 300 can be automatically refilled.
  • the movable arm 30 can be instructed to move the wash tip 41 (see Figure 10) on Z head 70 to the sponge cup position, and to pump sufficient water from the water reservoir through the water tubing and wash tip into the sponge cup.
  • the instruction set for controlling the movement of the Z head 70 can be programmed to ensure that the reagent tip head 40 is pressed onto the pipette tip 90 with sufficient force so as to form a tight seal therebetween, minimizing the risk -17- that the tip will be lost inadvertently during the operation of the apparatus.
  • O-ring 43, sponge cup 300, sponge cup support 301 and the O-ring fitting groove in reagent tip head 40 may be eliminated.
  • the operating station and function depicted in Figure 3 and Table 1 will be unnecessary.
  • Figure 4 is a detailed view of how the wash tip 41 and the blow tip 42 (as shown in greater detail in Figure 10) operate in the invention.
  • wash tip 41 (not shown in this figure) and blow tip 42 are integrated into Z head 70.
  • Z head 70 on movable arm 30 is moved upward so that Z head 70 is above any interfering part of the apparatus, then movable arm 30 is moved to an appropriate location while Z head is in the raised position.
  • Z head 70 reaches the appropriate location above a pre-selected microscope slide (position F), Z head 70 is again lowered to position wash tip 41 and blow tip 42 at an appropriate height above the selected microscope slide (as shown).
  • wash tip 41 and blow tip 42 are positioned at one end (e.g. the front end) of microscope slide 130 and a buffer or wash liquid, supplied through liquid supply conduit 62 (not visible in this figure), flows out from wash tip 41.
  • the Z head 70 is moved in a single pass to the rear of microscope slide 130. If desired, the blow operation can then be carried out on the same slide by supplying pressurized gas through an gas supply conduit to blow tip 42. The Z head 70 is then moved back to the front position of microscope slide 130 while keeping the gas stream from blow tip 42 in motion.
  • wash tip 41 and blow tip 42 to a second microscope slide for the addition of buffer, so that the buffer added to the first microscope slide can remain on the slide for a preselected period of time prior to removal.
  • wash tip 41 and blow tip 42 on Z head 70 are returned to the first slide of the group and the blow operation can commence.
  • wash tip 41 is provided with a bevel (therein depicted as approximately 30°) so that the surface tension of the (typically) aqueous liquid ordinarily dispensed through the tip will -18- prevent the formation of a depending drop of liquid which could loosen from the tip and contaminate a slide while the Z head 70 traverses the apparatus.
  • a bevel depicted as approximately 30°
  • FIG 5 is a front view of an embodiment shown generally in Figures 1 - 4, wherein the movable arm is in position 'D'.
  • the apparatus will ordinarily select a pipette tip.
  • the movable arm 30 will be directed to a location such that reagent tip head 40 is directly above and pointing to pipette tip 90 in pipette rack 92a or 92b positioned onto pipette tip holder 100.
  • the descending Z head 70 presses reagent tip head 40 into pipette tip 90 (here a disposable pipette tip), where the pipette tip 90 is retained on reagent tip head 40 by a press fit in the same manner in which tips are now retained on hand-operated pipettes.
  • Z head 70 is then raised and movable arm 30 relocated to position 'E' (as detailed in Figure 6).
  • pipette tip 90 mounted on reagent tip head 40 is lowered into a reagent vial 110 which is held in a predetermined location by reagent vial holder 120.
  • a supply of reagent is drawn into pipette tip 90 for application to microscope slides.
  • a measured volume of reagent can be withdrawn by a precise metering pump 37, which is driven by stepping motors to supply negative air pressure to reagent tip head 40 (i.e., withdrawing air through reagent tip head 40).
  • the application concept of a metering pump used herein is that the liquid located in the supply conduit 62 will act as a piston for withdrawing a specific volume of gas and thus drawing up a specific volume of reagent into pipette tip 90. According to experience and a well-defined conversion table (volume vs. number of steps), the accuracy of the volume can be controlled as appropriate.
  • FIG. 7 shows the details of the movable arm 30 and pipette tip 90 at the pre-selected position while applying the reagent to the slide by again using the stepping motor-driven metering pump 37 and the liquid retained in the supply conduit 62 to act as a piston.
  • Reagent can be applied to a single microscope slide 130, -19- or aliquots of the reagent in pipette tip 90 can be applied to different slides according to the calculation of the selected program instructions in the computer control system.
  • a tip ejection rod 64 is located on Z head 70, and is adapted to move along the Z-axis of the apparatus.
  • a tip ejection terminal block 63 is located at the side of Z head 70 above tip ejection rod 64. When it is desired to dispose of pipette tip 90, Z head 70 will be raised until the top end of tip ejection rod 64 contacts terminal block 63 installed on Z head 70.
  • tip ejection rod 64 Since further motion of tip ejection rod 64 is blocked by terminal block 63, while Z head 70 continues to rise, the lower end of tip ejection rod 64 will press against pipette tip 90 and force the tip off of the end of reagent tip head 40.
  • the pipette tip 90 then drops through tip disposal orifice 26, striking bar 25, and descending into tip disposal bin 27 (not shown) under framework 20 for later removal.
  • the reagent tip head 40 is then lowered to a normal position so that another pipette tip 90 can be mounted on reagent tip head 40.
  • the location assigned to pipette tip holder 100 is clearly separate from the location assigned to reagent vial holder 120.
  • these locations could be combined, for example, in an arrangement which stores a single pipette tip 90 in association with each reagent vial 110.
  • the reagent tip head 40 would be directed to the associated pipette tip 90 which would then be returned to the holder 100 for reuse after the reagent had been dispensed onto the microscope slide 130.
  • the control means of the apparatus 10 could monitor the volume of reagent remaining in each reagent vial 110, and arrange for disposal and replacement of the vial
  • FIG. 9 depicts three views of the movable arm 30, showing Z head 70 and reagent tip head 40 without a pipette tip 90 on the head.
  • the Z head 70 is mounted on the Z-axis track 39 and screw lead slide 38.
  • Z head 70 is -20- driven by a Z-axis motor (of which only the output gear 50 is shown in the horizontal front elevation of the figure).
  • a laser bar-code scanner 410 is located on the right side of the Z head 70.
  • a metering pump 37 which drives the reagent tip head 40 to withdraw and dispense reagent solutions.
  • the means to drive the metering pump can be, e.g., a two-phase stepping motor, desirably with approximately 1.8° of rotation per pulse.
  • a Z head latch 65 is installed on the left side of movable arm 30. When the apparatus 10 is shipped, the Z head latch 65 locks to the Z-axis track 39. The latch will release when Z head 70 returns to its home position.
  • the gas and liquid supply conduits and connection leads are not shown in the drawing, so that the major parts can be seen more clearly.
  • the movable arm 30 is mounted on the X-axis track 32 and can be directed linearly along the track under the direction of a computer or other means of control.
  • Figure 10 provides a more detailed view of the reagent tip head 40 on
  • reagent tip head 40 is depicted at the lower right side of Z head 70.
  • reagent tip head 40 has three different diameters at different cross-sections, namely: A segment 40a that is sufficiently large to act as a stop when press-fitting pipette tip 90 onto tip head 40; an intermediate section 40b that acts as the press-fit location for pipette tip 90; and a smaller-diameter segment 40c at the end of tip head 40.
  • an O-ring 43 is installed and inserted into a groove on the smaller-diameter segment 40c.
  • the outer diameter of O-ring 43 is selected so that when pipette tip 90 is pressed onto tip head 40, O-ring 43 will seal the space between tip head 40 and pipette tip 90.
  • the O-ring 43 thus engaged helps to eliminate leakage and increase the friction fit of pipette tip 90, thereby -21- securing the retention of pipette tip 90; a feature of benefit where pipette tip 90 may suffer from some degree of shape distortion.
  • the instruction set for controlling the movement of the Z head 70 can be programmed to ensure that the reagent tip head 40 is pressed onto the pipette tip 90 with sufficient force so as to form a tight seal and thus minimize the risk that the tip will be lost inadvertently during the operation of the apparatus.
  • Figure 11 shows a series of views of wash tip 41 and blow tip 42.
  • On the left of the figure is a sectional view of Z head 70, depicting three vertical channels in this embodiment.
  • the left channel is provided for wash tip 41
  • the middle channel is for blow tip 42
  • the right channel is for reagent tip head 40.
  • the reagent tip head 40 is not shown in the figure, for clarity of presentation of the remaining features.
  • the hollow interior of blow tip 42 is not limited to the specific shape shown, although certain advantages are obtained for aspects of this interior shape and slit shape as described below.
  • the hollow interior 77 of the blow tip 42 can vary significantly in shape as long as sufficient access is provided for easy flow of gas into interior space 77 so that pressure differentials do not build up and cause differential exit of gas through exit slot 79.
  • Exit slot 79 is generally located on the bottom-most surface of blow tip 42 and is preferably a linear exit slit having a length substantially equal to the width of a standard microscope slide of the type selected for use in a particular operation. As there are different sizes of microscope slides, different wash and blow tips can be prepared for the dimensions of each such microscope slide. As shown in the sectional view C of Figure 11, access 78 between hollow interior space 77 and exit slit 79 is preferably provided so that gas leaving slit 79 exits at an angle to the vertical. By providing exit slit 79 at a slight angle and moving Z head 70 in a direction toward the obtuse angle formed between the gas wall and the microscope slide 130, the removal of water or buffer from slide 130 is facilitated.
  • FIG. 11 At the right side of Figure 11 is a sectional view of a liquid distribution embodiment of the invention. There is portrayed a main vertically-oriented hollow located at the center of the embodiment. An opening 76 shown at the top of the embodiment is used during the manufacture of the device, and will ordinarily be -22- blocked after the installation is completed. However, opening 76 can also be used as an inlet for an extra liquid supply. Wash tip 41 will be installed at the lower end of the embodiment, and is not shown in the figure. On the side wall of the embodiment, there is a row of liquid solution inlet connectors. This array of parallel channels provides a path for the incoming liquid solution to be distributed to the main hollow portion, which can then flow down to wash tip 41.
  • FIG. 12 shows several typical reagent dispensing patterns on a microscope slide 130. Most slides will have a specimen region 132 and a second region 134 for including information on the slide.
  • a specimen can be present at any location within region 132.
  • the thin film of buffer that will be present when a reagent is dispensed onto the slide assists in ensuring that adequate reagent is applied to the specimen, regardless of where the specimen is located or the reagent is applied.
  • the reagent is dispensed in a pattern, rather than a single location, so that the distance that a reagent must diffuse through the liquid film is reduced.
  • the 1/3 and 2/3 patterns depicted in Figure 12 restrict dispensed reagent to a portion of region 132, in order to conserve reagents if a specimen occupies only a limited portion of the region. This can prove valuable where the reagents are expensive, such as, e.g., nucleic acid probes.
  • Special slides are available commercially which include pre-established fluid barriers, or the user can establish such barriers by creating a border around the specimen with a hydrophobic material, such as a marker (PAP) pen, also available commercially.
  • PAP marker
  • Figure 13 shows multiple views of a tray 190 intended to hold microscope slides.
  • Tray 190 is formed into a series of individual wells 192 for microscope slides; the location of a single microscope slide 130 is shown by a dotted line in the right-most well of the plan view in Figure 13.
  • the design of the well fits both 1" x 3" U.S. standard and 26 x 76mm European standard microscope slides.
  • Individual side walls 194 separate each well 192 from its adjacent wells to prevent accidental contact of liquid, such as might occur during a washing operation, and to prevent contamination between adjacent microscope slides.
  • the open bottom of each well 192 allows buffer to drain through the bottom of tray 190 where it will be disposed of, typically in a drain bin 27 as shown in Figure 2.
  • the open bottom of the well 192 will also permit the use of projections on the baseplate 22 to raise the slide 130 above the tray 190, so that the edges of the slide 130 in the region where reagent is applied are not in contact with the tray 190, thus precluding a capillary wicking of reagent or other solution and possible cross-contamination of the slide.
  • the side walls 194, retaining tabs 196 and bracing feet 197 closely and accurately retain microscope slides placed into the individual wells.
  • a gap 198 is present at one end of well 192 to allow easy grasping of an individual microscope slide 130 between thumb and forefinger for insertion into and removal from tray 190.
  • the blow tip 42, wash tip 41 and reagent tip head 40 are integrated onto Z head 70 as one embodiment.
  • the blow tip head 42 when performing a blow operation, is located very close to the surface of slide 130 which is set into the slide tray 190, and the reagent tip head 40 at the same height is positioned just in front of blow tip 42.
  • a row of head openings 199 at the front wall is designed to provide a space for reagent tip head 40 when blow tip 42 moves to the rear end of the slide 130.
  • Removable trays 190 are designed for ease of operation by allowing a user to place microscope slides 130 in a loading tray 190 outside the apparatus 10 in which the staining operations will occur.
  • Another -24- advantage of allowing trays 190 to be removed from the slide tray holder is to simplify the process of cleaning and other maintenance work.
  • the tray 190 also will be adapted to fit precisely into other elements at the appropriate location on base plate 22.
  • right tray support 206 and left tray support 207 which are installed on the baseplate 22, contain four slide trays 190 in a row, each of which is capable of containing 10 slides for a total of 40 microscope slides in a predetermined array.
  • Figure 14 shows how a tray 190 is set onto tray supports 206/207.
  • on right tray support 206 there are four sliding rods 195 installed horizontally, facing and against the outside wall on the right side of the tray 190.
  • the another end of sliding rod 195 (right side) is linked with a micro lever switch which is in turn electrically connected to a in-position light 191.
  • the sliding rod 195 can freely move left or right.
  • a detailed view is shown in Figure 14.
  • the sliding rod 195 When there is no tray on the tray support, the sliding rod 195 is at the left-most position, so that switch 193 is 'off and the light 191 is likewise off.
  • the right side wall of the tray 190 pushes sliding rod 195 to the right, pressing the micro lever switch to its 'on' position, then activating the light 191.
  • the light 191 then indicates that the tray 190 is now in the correct position and ready for staining.
  • Another function for sliding rods 195 is to 'soft lock' the trays 190 and prevent them from moving when the apparatus 10 is in operation. In using such a sensing mechanism, care should be taken to prevent contamination from spilled liquids or particulates which could obscure the light path from light 191, thereby falsely indicating the presence of a loaded rack.
  • Figure 14 also depicts the slide location bar-code 400 which can be presented on strips associated with the baseplate 22, the tray supports 206/207 or with the microscope slide tray 190.
  • This bar-code 400 will aid the control means in associating a selected slide 130 with the proper reagent vial 110 and staining protocol. While the relative positions of microscope slides 130 in the present embodiment can be established by scanning only three strips of bar-code 400, as a practical matter there is no appreciable loss of efficiency in utilizing a bar-code to identify the location of every slide in the apparatus. -25-
  • reagent vial holder 120 located at the left side of slide tray support 207 is designed to hold up to 40 reagent vials and can accept reagent vials having a wide range of capacities, typically from 5mL to 25mL.
  • a vial holder block (not shown) having appropriate dimensions could be utilized in place of the depicted vials.
  • Such a block could include a bar-code and present a smaller vial, such as a screw-cap vial commonly used to contain small volumes of expensive reagents, in proper orientation for access by the reagent tip head 40.
  • Such a holder block can thus extend the useful volume range as low as is practical, typically as low as OJmL, a range commonly associated with nucleic acid reagents.
  • a detailed view of one embodiment of reagent vial holder 120 is shown in
  • FIG 15. The total of 40 reagent vials are divided into two columns with 20 vials in each.
  • the left column of reagent vial holder 120 contains reagent vials #1 to #20 starting from the upper left corner of reagent vial holder 120 (see Figure 1) and down to vial #20 in the same column in vial holder 120.
  • the right column of reagent vial holder 120 holds reagent vials #21 to #40.
  • the vial numbers can be engraved right at the position where the reagent vial will be located. These numbers correspond to the specific reagent vial numbers in the programming of the instrument.
  • the spacing prongs 122 between two reagent vials are designed to maintain certain spacing between adjacent vials, and accommodate shape/size distortions of the plastic reagent vials 110. In this manner, the reagent vials will be secure in the proper positions.
  • Handles 124 can be installed at both ends of reagent vial holder 120 to simplify manipulation.
  • a reagent location indicator 403 is mounted on left slide tray support 207. Hinges are used to allow the location indicator 403 to be repositioned when loading the reagent vials 110 onto vial holder 120; and the location indicator 403 can be returned to cover the tops of the reagent vials after loading.
  • This reagent location indicator 403 resembles a 'fish bone,' with a 'spine' in the middle and total of 38 'fins' evenly spaced on both side of the 'spine. ' A bar-code and a human readable number is printed on each 'fin' to indicate the reagent vial number.
  • the indicator 403 When in working position, the indicator 403 is in a horizontal orientation laying on the tops of -26- the reagent vials 110.
  • the 'spine' is then located at the center divider of two columns of the reagent vials; each fin is positioned between two adjacent reagent vials in a column, and the bar-codes on the "fins" are 'inserted' between the bar-codes on the reagent vials 110.
  • the reagent vial 110 will also be configured so as to provide a region 112 for the bar-code label in a horizontal orientation on the upper surface of the vial. In this manner, the reagent vial information can be readily accessed at the same time as the location information from indicator 403.
  • Figure 16 shows various components used in the supply of gas and wash liquids to Z head 70.
  • An gas compressor 170 provides gas through flexible conduit 172 to movable arm 30, Z head 70 and ultimately to blow tip 42.
  • a computer controlled switch controls gas pump on and off to control the gas supply.
  • Buffer or other wash solutions are supplied by individual pulse metering pumps. The volume of liquid is controlled by the number of pulses sent to the metering pump.
  • Pump 173 is for Dl water
  • pump 174 is for Buffer solution
  • pump 175 is used for pumping dewaxing solutions.
  • Different liquid solution are pumped from individual reservoirs through individual supply tubing conduits 62, movable arm 30, Z head 70, and finally to the common wash tip 41.
  • the material used for supply conduit tubing must be selected individually for specific liquid solutions.
  • the tubing used for dewaxing solutions should be resistant to organic solvents and detergents, for example Viton-type tubing.
  • the present apparatus enables the inclusion of a dewaxing slide preparation step in the automated protocols implemented by the control system. This is a feature previously unavailable in apparatus which process microscope slides in a horizontal orientation. Representative of protocols useful in such slide preparation protocols are the methods disclosed in PCT Publication WO 95/24498 and in U.S. -27-
  • FIG. 17 depicts a typical trail that the laser bar-code scanner 410 will trace when reading the bar-codes on both microscope slides 130 and reagent vials 110.
  • a batch processing method is generally used in bar-code reading in the present invention. Rather than reading the bar-codes and acting on the information individually, the laser scanner 410 scans the bar-codes by batch in order to increase processing speed. In a typical pattern, laser scanner 410 begins scanning at the left most slide 130 in tray #1
  • the scanner 410 on the movable arm 30 moves vertically down to the bottom row (tray #4), the scanner reading the bar-codes 402 on slide #1, #11, #21 and #31, and each location indicator bar-code 400 which is in the same column as well.
  • the information can be saved in laser scanner 410 as it scans, then transferred to the computer, e.g. via a RS232 serial port, using a protocol compatible for both. The most commonly used protocol in this regard is 7 bit data, 1 bit stop and even parity check.
  • Movable arm 30 then moves one slide left, where laser scanner 410 is aiming at the microscope slides in the next column (#2, #12, #22 and #32).
  • slide location indicator 400 (with associated bar-code) can be installed on each slide tray (optionally except row #1 or #4).
  • the laser scanner 410 also reads the bar-code on indicator 400 between the two bar-codes on the slides.
  • the computer is capable of identifying which slide is "missing" and a menu on the computer screen informs the user to manually input the missing information or to re-run the scanning procedure.
  • movable arm 30 moves laser scanner 410 to the reagent vials -28-
  • the apparatus can process trays of slides in a manner which completes the prescribed processing on a single tray 190, and then signal the user to remove the tray and replace it with a fresh tray, without interrupting the processing of the remaining trays. In this manner, the apparatus can be utilized continuously with a minimum of intervention by the user.
  • the X, Y, and Z tracks can be replaced by a single robotics arm. Additionally, it will be understood that the specific tracks, motors and other individual parts can be replaced by other parts of equivalent function.
  • the X, Y, and Z tracks are purchased as commercially available sliding tracks.
  • the X-axis and Z-axis motions are supplied by a linear motion rail assembly and linear motion guide (Part Nos. LWES 20C1 R690
  • the Y-axis shaft system is a linear bearing rail assembly (Part No. SRA-8-PD-21-55 also supplied by Specialty Motions); power along the Y-axis was supplied by the same stepping motor used for the X-axis.
  • the apparatus of the invention can be prepared from readily available commercial parts assembled in the manner described, with a minimum number of specialized manufacturing techniques. Since the Z head 70 is not readily available, it will typically be manufactured for a particular apparatus as shown or in a similar manner to provide the features that are described in the specification above.
  • the composition of the components from which various parts are manufactured can vary widely, but components through which reagents pass or which contact -29- potentially corrosive reagent or wash solutions are typically prepared from stainless steel or inert plastics to prevent corrosion.
  • the Z head 70 with its integrated tips and other features is typically formed from a moldable plastic (such as a polyacrylate) and can be prepared by a molding process, a plastic-shaping process, or some combination thereof depending on the individual shape intended to be utilized. Parts that are subject to wear, such as the stem 38 and reagent tip head 40 of Z head 70 are typically prepared from a hard plastic or other material that will resist wear.
  • the present apparatus is typically operated under the control of a computer or other programmable control device.
  • a computer or other programmable control device In the simplest applications, where only a single type of automated staining well be performed repeatedly, it is possible to provide either a hard- wired controller or a non-programmable electronic controller, such as a computer operating under instructions from read-only memory. In preferred embodiments, however, a programmable controller or computer is used so that the operation of the apparatus can be varied. Software will generally be provided with the computer so that the user does not need to provide instructions for individual motions, but merely selects appropriate motions from a menu.
  • bar-code technology can be used to supply instructions to the apparatus.
  • the apparatus reads bar-codes associated with both the reagent vials 110 and the slides 130; thereafter the computer is able to determine all parameters needed to carry out the most appropriate pre-programmed instruction set in the memory of the computer to control the apparatus in the processing procedures for microscope slide staining.
  • less user input is required, thus reducing the opportunities for introduction of error.
  • This feature will avoid lost steps, provide more accurate positioning and help to reduce "dead volume” (i.e. less reagent remaining at the bottom of a reagent vial. It is also possible to provide within the instruction set provisions for recovery from a loss of pipette tip during a staining routine, as well as machine calibration.
  • One desirable step in the method that is used in the automated apparatus of the invention involves blowing excess reagent or buffer off the surface of the slide 130. A preferred embodiment of the tip used in this blowing operation is shown in Figure 11, although other tips having slits for exit of gas to provide a wall of gas can also be used.
  • the extent to which liquid is removed from the slide can be varied.
  • the amount of liquid present in the thin film on the microscope slide's upper surface is quite small, typically from 2 to 25 microliters, more generally from 3 to 20, and preferably from 5 to 10.
  • the area to be covered is generally approximately 15cm 2 , providing a typical volume per surface area of approximately 0J3 to 1.7 microliter/cm 2 .
  • it is difficult to determine the actual volume being used since the operation of blowing liquid off the top surface of a slide causes liquid to adhere to other portions of the microscope slide, making measurement of the remaining liquid difficult.
  • the volume of liquid present on the slide upper surface at the end of a blow operation is best determined empirically.
  • the maximum permissible volume is determined by the stain being used and its concentration at the reagent application -31- stage, since these factors affect the final concentration of the stain or other reagent on the surface of the slide.
  • Historical procedures developed for slide preparation are generally described in terms of a particular reagent concentration, incubation time, and temperature. Accordingly, it is desirable to provide a minimum volume of liquid on the slide in order to avoid having to change the concentration of reagents from the standard used in the industry. By adhering to this guideline, it is possible to use commercially available, ready-prepared stain solutions as reagents.
  • the gas pressure, height of the head above the slide and rate of motion of the head for control of the liquid film can all be selected by the user or by the manufacturer of the apparatus. Generally, the same gas pressure will be used at all times so as to remove this variable from consideration. Thus, only the height of the head and the rate of motion will typically be varied. The higher the head above the slide, the less liquid will be removed. The faster the head is passed across the slide for a given height, the less liquid will be removed.
  • an gas pressure of 7psi (0.5cm/sec), a height of 0.07 inch (2mm) above the microscope slide surface, and a rate of motion of 3 inch/sec (7.5cm/sec) provide a preferred buffer film suitable for the staining of four slides at 25 °C and a relative humidity of 60-80% , which is the typical humidity present inside a closed and operating apparatus of the invention.
  • wash solutions used in the apparatus of the invention can vary significantly depending on the staining technique being used.
  • a typical wash solution is an aqueous solution of a surfactant and can contain other components present in typical slide preparation of wash solutions, such as buffers.
  • sufficient surfactant is present to provide a surface tension in a solution equivalent to that present in solutions containing water and the following surfactants at the concentrations listed.
  • Typical surfactants used are TWEENTM 20 (0.02 to 2% v/v), BRIJTM35 (0.05 TO 3% v/v), and TRITONTM X-100 (0.01 to 1 % v/v).
  • Typical buffers used are phosphate buffered saline and TRIS-Cl (each at approximate pH 7.6).
  • water can and generally is an aqueous solution of buffer and surfactant or of some staining reagent.
  • the apparatus of the invention can be used in any staining technique that can be carried out manually and that there are no limitations placed on the invention by the staining technique.
  • the apparatus of the invention can contain a number of further components designed for ease of operation.
  • drain trays with exit conduits to waste reservoirs can be located either individually under components of the apparatus or a single drain tray and collection system can be provided for the entire interior space of the apparatus frame.
  • the framework is a form of a cabinet with an interior space in which all operations take place.
  • a closeable access port e.g., a door
  • a transparent door can be provided to prevent accidental spraying of liquid (as during a blowing operation) into the room in which the apparatus is located, while allowing the user of the apparatus to visually verify proper operation.
  • kits intended to ensure level operation, to protect against electric shock, to verify that an appropriate tip has been selected and properly placed on the tip head, or to optically scan slides in a microscope slide tray or other container for microscope -33- slides so that a user is not required to enter information into the computer.
  • Such information could be provided, for example, by a standard bar-code attached to an individual microscope slide or the component.
  • Multiple reagent containers can be provided so that different staining operations can be carried out under the control of the bar-code system and the computer and its pre-programed software.
  • Ventana 320 Automated immunostaining system Marketing brochure for Ventana Medical System, Inc., Arlington, AZ (date unknown).

Abstract

L'invention se rapporte à un dispositif automatisé de coloration comportant un bras (30) susceptible de se déplacer suivant trois dimensions, et une tête d'extrémité creuse (70) disposée sur le bras et comportant une tête à pointe pour réactif (40), une pointe de lavage (41) et une pointe de soufflage (42) qui permettent de répartir sélectivement un gaz et un liquide sur des lames porte-objet. L'invention se rapporte également à divers accessoires dudit appareil qui sont conçus particulièrement pour le traitement de spécimens sur des lames porte-objet.
PCT/US1998/005919 1998-03-24 1998-03-24 Appareil automatise de coloration WO1999049295A1 (fr)

Priority Applications (6)

Application Number Priority Date Filing Date Title
AU69417/98A AU762808B2 (en) 1998-03-24 1998-03-24 Automated staining apparatus
US09/646,695 US6495106B1 (en) 1998-03-24 1998-03-24 Automated staining apparatus
PCT/US1998/005919 WO1999049295A1 (fr) 1998-03-24 1998-03-24 Appareil automatise de coloration
CA002325583A CA2325583A1 (fr) 1998-03-24 1998-03-24 Appareil automatise de coloration
EP98915168A EP1066502A1 (fr) 1998-03-24 1998-03-24 Appareil automatise de coloration
JP2000538216A JP2002507738A (ja) 1998-03-24 1998-03-24 自動着色装置

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/US1998/005919 WO1999049295A1 (fr) 1998-03-24 1998-03-24 Appareil automatise de coloration

Publications (1)

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WO1999049295A1 true WO1999049295A1 (fr) 1999-09-30

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EP (1) EP1066502A1 (fr)
JP (1) JP2002507738A (fr)
AU (1) AU762808B2 (fr)
CA (1) CA2325583A1 (fr)
WO (1) WO1999049295A1 (fr)

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CA2325583A1 (fr) 1999-09-30
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AU6941798A (en) 1999-10-18
EP1066502A1 (fr) 2001-01-10

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