WO2011002779A2 - Procédés et appareils d'application et de retrait de fluides pour traiter des échantillons biologiques - Google Patents

Procédés et appareils d'application et de retrait de fluides pour traiter des échantillons biologiques Download PDF

Info

Publication number
WO2011002779A2
WO2011002779A2 PCT/US2010/040413 US2010040413W WO2011002779A2 WO 2011002779 A2 WO2011002779 A2 WO 2011002779A2 US 2010040413 W US2010040413 W US 2010040413W WO 2011002779 A2 WO2011002779 A2 WO 2011002779A2
Authority
WO
WIPO (PCT)
Prior art keywords
slide
substrate
liquid
gap
holder
Prior art date
Application number
PCT/US2010/040413
Other languages
English (en)
Other versions
WO2011002779A3 (fr
Inventor
Matthew David Mette
Timothy James Keller
Kevin David Marshall
Michael Otter
Original Assignee
Ventana Medical Systems, Inc.
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 Ventana Medical Systems, Inc. filed Critical Ventana Medical Systems, Inc.
Publication of WO2011002779A2 publication Critical patent/WO2011002779A2/fr
Publication of WO2011002779A3 publication Critical patent/WO2011002779A3/fr

Links

Classifications

    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/34—Microscope slides, e.g. mounting specimens on microscope slides
    • 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
    • G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on

Definitions

  • the present invention relates generally to methods and apparatuses for processing samples. More specifically, the invention is related to methods and apparatuses for applying and removing fluids for processing biological samples.
  • Example techniques include microscopy, micro-array analyses (e.g., protein and nucleic acid micro-array analyses), and mass spectrometric methods.
  • Samples are prepared for analysis by applying one or more liquids to the samples. If a sample is treated with multiple liquids, both the application and subsequent removal of each of the liquids can be important for producing samples suitable for analysis.
  • Microscope slides bearing biological samples e.g., tissue sections or cells
  • Samples can be prepared for analysis by manually immersing sample-bearing slides in containers of dyes or other reagents.
  • This labor intensive process often results in inconsistent processing and carryover of liquids between containers. Carryover of liquids leads to contamination and degradation of the processing liquids.
  • These types of manual processes often utilize excessive volumes of liquids resulting in relatively high processing costs, especially if the dyes or other reagents are expensive and are prone to degradation due to carryover.
  • “Dip and dunk” automated machines immerse samples in liquids similar to manual immersing techniques. These automated machines can process samples in batches by submerging racks carrying microscope slides in open baths. Unfortunately, relatively large amounts of reagents are in bath containers of the dip and dunk automated machines. Similar to manual processes, if the liquids are expensive reagents, processing costs may be relatively high, especially if significant amounts of reagents are wasted. Reagent bath containers may be frequently emptied because of contamination due to carryover. Open containers are also prone to evaporative losses that may significantly alter the concentration of the reagents resulting in inconsistent processing. It may be difficult to process samples without producing significant volumes of waste that may require special handling and disposal.
  • Wicking techniques can be used to wick out excess reagents.
  • Wicking material e.g., a paper towel or other absorbent material
  • a glass coverslip can be mounted on the slide to protect tissue from drying and fading.
  • a paper towel is placed under or alongside an edge of the coverslip. The paper towel wicks excessive liquid between the coverslip and the slide.
  • clean wicking materials are used for each slide to prevent contamination, resulting in the accumulation of used wicking materials and significant disposal costs.
  • At least some embodiments disclosed herein are directed to slide processing apparatuses that are capable of using a coverslip to apply a liquid to a sample carried on a microscope slide. Capillary forces provided by the liquid pull the coverslip to help distribute the fluid evenly.
  • An actuator assembly can move the coverslip relative to the slide. Capillary forces and forces applied by the actuator assembly are balanced to avoid overwetting, over-filling, and/or underfilling.
  • the slide processing apparatus provides positioning (e.g., self- alignment, self-gapping, or the like) of the coverslip and the microscope slide and may be generally insensitive to processing volumes and/or evaporation.
  • the actuation assembly includes at least one flexure and a counterweight spring that cooperate to provide a force that prevents overwetting while allowing self-alignment and/or self-gapping.
  • the flexure and spring cooperate to keep the coverslip floating on the liquid.
  • the flexure can provide a force in the opposite direction to maintain a minimum distance between the coverslip and the microscope slide.
  • the flexure provides a sufficient force to maintain a nominal height of the gap in a range of about 0.003 inch to about 0.005 inch for about 100 micro liters of liquid.
  • the coverslip can remain generally static to form an even liquid layer to enhance and help ensure uniform reagent uptake by a sample.
  • the coverslip is moved with respect to the microscope slide to move the liquid to a desired location.
  • a holder holding the coverslip can be moved away from and towards the microscope slide to agitate the liquid.
  • the coverslip is rocked back and forth to move the liquid along the gap using capillary action.
  • the microscope slide and coverslip are rotated together to cause movement of the liquid along the gap.
  • the substrate and coverslip are moved from an inclined orientation for receiving the liquid to another orientation to cause the liquid to flow along the gap.
  • the microscope slide and coverslip can be rotated about an axis of rotation.
  • a portion of the coverslip can be moved away from the microscope slide to urge the liquid towards a narrowed region of the gap.
  • the liquid is aspirated from the gap using a contactless vacuum nozzle.
  • the narrowed region is widened to lower the capillary forces holding the liquid in the gap and then the liquid is aspirated from the gap.
  • the narrowed region is widened at least about 0.005 inch, 0.01 inch, 0.015 inch, or 0.02 inch. Another liquid can then be delivered into the gap for further processing of the sample and/or coverslipping.
  • an apparatus for delivering a reagent onto a slide includes a slide handler and a waste remover.
  • the slide handler is configured to hold a slide carrying a specimen.
  • the cover handler opposes the slide handler and includes a holder and an actuation assembly.
  • the holder is adapted to hold and release the cover.
  • the actuation assembly is operable to move the cover held by the holder with respect to the slide to form a variable height capillary gap to cause movement of the reagent along the variable height capillary gap while keeping the cover spaced apart from the slide.
  • the waste remover includes an aspiration port positioned to remove the reagent in the capillary gap.
  • an apparatus for applying a liquid to a specimen carried on a slide includes a slide handler and a cover handler.
  • the slide handler is configured to hold a slide carrying a specimen.
  • the cover handler faces the slide handler and includes a holder and an actuation assembly.
  • the actuation assembly is operable to drive the holder away from and towards the slide handler and is operable to allow capillary forces produced by a liquid in a capillary gap between the slide held by the slide handler and the cover held by the holder to align the cover with the slide.
  • the actuation assembly and the holder allow the cover to translate and rotate with respect to the slide in response to forces provided by the liquid.
  • the actuation assembly includes a biasing mechanism configured to provide a biasing force that is less than the capillary force produced by the liquid.
  • the biasing mechanism includes a flexure that is substantially parallel to the holder.
  • a substrate holder for positioning a substrate along a slide includes a main body, a flexure coupled to the main body, a holder, and at least one actuation mechanism coupled to the holder.
  • the holder is suspended by the flexure.
  • the holder is configured to hold the substrate while the substrate distributes a liquid along a microscope slide.
  • the actuation mechanism is movable to position the holder so as to define a capillary gap between the substrate and the slide to cause movement of a liquid along the capillary gap while keeping the substrate spaced apart from the slide.
  • the capillary gap has a variable height.
  • the holder includes a holder base and a permeable member coupleable to the holder base such that the microscope slide is held against a face of the permeable member when a vacuum is applied to the permeable member via the holder base.
  • the flexure is substantially parallel to a substrate contact face of the holder. The flexure is sufficiently flexible to enable translation and rotation of the holder with respect to the main body in response to capillary forces provided by the liquid in the capillary gap.
  • the actuation mechanism includes a first actuator positioned to move a first end of the holder and a second actuator positioned to move a second end of the holder.
  • the second end of the holder opposes the first end of the holder.
  • the first actuator and the second actuator are operable to independently move the first end and the second end of the holder away from and towards the slide.
  • the flexure can be extendable to allow the holder to move towards the slide, and the flexure is contractable to move the holder away from the slide.
  • the flexure can include at least three arms that extend outwardly from a retainer of the holder. Each arm is distensible for side-to-side movement of the holder and is bendable for rotating the holder.
  • an apparatus for positioning a first substrate with respect to a second substrate includes a substrate holder and an actuation assembly.
  • the actuation assembly includes an actuation mode and a self-alignment mode.
  • the actuation assembly moves a first substrate held by the substrate holder with respect to a second substrate when in the actuation mode.
  • the actuation assembly allows substantially free movement of the first substrate due to forces provided by a liquid in a capillary space between the first substrate and the second when in the self-alignment mode.
  • the actuation assembly in certain embodiments, includes at least two actuators and a flexure.
  • the actuators are connected to the substrate holder to translate and rotate the second substrate and the flexure sufficiently compliant to enable the second substrate to float on the liquid in the capillary gap.
  • the actuation assembly includes at least one linear actuator for displacing one end of the second substrate to rotate the second substrate without contacting the first substrate with the second substrate.
  • a method comprises positioning a microscope slide and a substrate at a receiving orientation.
  • a liquid is delivered between the microscope slide and the substrate.
  • the microscope slide and substrate are moved from the receiving orientation to another orientation to move the liquid along a gap defined by the microscope slide and the substrate.
  • the microscope slide and substrate slope upwardly at an angle of inclination greater than about 10 degrees and less than about 80 degrees while the liquid is delivered.
  • the microscope slide and substrate slip upwardly at an angle of inclination of about 45 degrees to about 55 degrees. To agitate the liquid, the microscope slide and substrate can be inverted.
  • a method of applying a liquid to a slide comprises delivering a liquid between a first substrate and a second substrate.
  • the first substrate carries a specimen.
  • the second substrate is moved with respect to the first substrate to accumulate the liquid in a gap defined by the first substrate and the second substrate using capillary action.
  • the accumulated liquid is aspirated from the gap.
  • mounting media is delivered between the first and second substrates.
  • the method in certain embodiments, involves using mounting media comprising an adhesive.
  • the adhesive is cured to permanently couple the first substrate to the second substrate.
  • the adhesive comprises one or more thermosetting materials.
  • Delivering the liquid can include passing the liquid through an opening formed by an end of the second substrate and a face of the first substrate.
  • Moving the second substrate with respect to the first substrate can include allowing the second substrate to move to a substantially parallel orientation with respect to the first substrate in response to capillary forces provided by the liquid in the gap.
  • the liquid can be allowed to equilibrate to provide a substantially uniform layer of the liquid along substantially an entire length of the gap.
  • the method can further include floating the second substrate on the liquid while a chuck holds the first substrate via a vacuum.
  • Aspirating the accumulated liquid may include contactlessly removing the accumulated liquid by drawing the accumulated liquid out of the gap and into an inlet of a waste remover. The inlet is spaced apart from both the first substrate and the second substrate.
  • a generally uniform height layer of the liquid can be formed by overlaying the first substrate held by a first holder with the second substrate held by a second holder to spread the liquid via capillary action such that the liquid fills most of the gap and pulls the second substrate held by the second holder towards the first substrate held by the first holder.
  • the second substrate is moved with respect to the first substrate by moving a portion of the second substrate away from an opposing portion of the first substrate to move the liquid towards a narrowed region of the gap.
  • the substrates can be moved by translating the second substrate away from the first substrate and rotating the second substrate with respect to the first substrate.
  • a minimum distance between the first substrate and the second substrate can be maintained. The minimum distance can be kept at a distance that is greater than about 0.004 inch while removing a substantial volume of the accumulated liquid.
  • At least a portion of the gap can be widened to move the liquid to a relatively narrow section of the gap via capillary action.
  • a method comprises delivering a liquid between a slide held by a slide holder and a cover held by a cover holder.
  • the cover is moved to cause accumulation of the liquid at a region of a gap proximate to an aspiration port using capillary action.
  • the gap is formed by the slide and the cover.
  • the accumulated liquid is removed from the gap.
  • mounting media is delivered between the slide and the cover. After delivering the mounting media, the cover is separated from the cover holder.
  • the method in some embodiments, further comprises curing the mounting media to permanently couple the cover to the slide.
  • the mounting media can comprise, in whole or in part, a light curable adhesive.
  • the cover can be moved relative to the slide to fill most of the gap by volume with the mounting media.
  • the cover in some embodiments, is moved to allow capillary forces provided by the liquid to position the cover with respect to the slide.
  • a method of applying a reagent to a slide includes altematingly delivering and removing reagents to a specimen carried on a slide using a substrate overlaying the specimen and a mounting region of the slide. After altematingly delivering and removing the reagents, delivering an adhesive between the mounting region and the substrate.
  • reagents can be successively applied to the specimen to perform a desired protocol.
  • the method further comprises holding the slide with a slide holder and holding the substrate with a substrate holder while alternatively delivering and removing the reagents.
  • the specimen is stained by altematingly delivering and removing the reagents.
  • Altematingly delivering and removing reagents includes successively delivering a respective one of the reagents from a respective pipette and aspirating the respective one of the reagents from between the slide and the substrate.
  • a method of applying a liquid to a biological sample includes providing a biological sample on a microscope slide.
  • a liquid is delivered between the microscope slide and a substrate.
  • a layer of the liquid is formed in a gap defined by a surface of the microscope slide and an opposing surface of the substrate such that the layer of liquid contacts the biological sample.
  • the liquid is accumulated at a region of the gap at least proximate to an opening of the gap via capillary action. The accumulated liquid is contactlessly moved through the opening out of the gap.
  • the substrate can float on the liquid to cause the surface of the substrate to become substantially parallel to the surface of the microscope slide.
  • a portion of the substrate can be moved away from an opposing portion of the microscope slide to move the liquid towards the region of the gap. After accumulating the liquid at the region of the gap, the gap is widened to reduce capillary forces pulling the substrate towards the microscope slide. The accumulated liquid is thus contactlessly removed after widening the region of the gap.
  • a minimum distance can be maintained between the microscope slide and the substrate greater than about 0.001 inch while accumulating the liquid at the region of the gap and while contactlessly removing at least a portion of the accumulated liquid. Substantially the entire microscope slide is spaced apart from the substrate prior to contactlessly removing the accumulated liquid.
  • a method comprises positioning a microscope slide and a substrate at an inclined orientation.
  • a liquid is delivered between the microscope slide and the substrate.
  • the microscope slide and the substrate are moved from the inclined orientation to another orientation to move the liquid along a gap defined by the microscope slide and the substrate.
  • the microscope slide and the substrate can slope upwardly at an angle of inclination greater than about 10 degrees and less than about 80 degrees while delivering the liquid.
  • the slide and substrate can be inverted.
  • the microscope slide and the substrate can be rotated about an axis of rotation at least about 45 degrees.
  • the liquid can be agitated by rotating the microscope slide and the substrate.
  • Figures 1-4 are side elevational views of a pair of substrates processing a sample.
  • Figure 5 is an isometric view of a slide processing apparatus capable of applying fluids to a sample carried on a slide and removing waste from the slide, in accordance with one embodiment.
  • Figure 6 is a side elevational view of the slide processing apparatus of Figure 5 with a bolus of fluid on the slide.
  • the bolus of fluid is ready to be removed by a waste remover.
  • Figure 7 is a detailed view of the bolus of fluid in a gap between the slide and an opposable substrate.
  • Figure 8 is a side elevational view of the fluid being drawn across a gap and into the waste remover.
  • Figure 9 shows a capillary gap between the slide and opposable substrate after the fluid has been removed.
  • Figure 10 is a side elevational view of a substrate holder delivering a processing fluid onto a sample carried by a slide, in accordance with one embodiment.
  • Figure 11 is a side elevational view of the processing fluid of Figure 10 spread along a capillary gap.
  • Figure 12 is a side elevational view of the substrate holder of Figure 10 with a substrate angled with respect to a slide to accumulate the processing fluid at one end of the capillary gap.
  • Figure 13 is a side elevational view of the substrate holder of Figure 10 with the substrate angled with respect to the slide to accumulate the processing fluid proximate to a waste remover.
  • Figure 14 is a side elevational view of the substrate holder of Figure 10 with the processing fluid accumulated proximate to an inlet of the waste remover.
  • Figure 15 is a side elevational view of a slide processing apparatus capable of rotating a slide, in accordance with one embodiment.
  • Figure 16 is a side elevational view of the slide processing apparatus of Figure 15 holding the slide at an inclined orientation.
  • Figure 17 is a partial cross-sectional view of a slide processing apparatus ready to dispense a processing fluid onto a sample.
  • Figure 18 is a partial cross-sectional view of the slide processing apparatus of Figure 17 with the processing fluid spread along an opposable substrate.
  • Figure 19 is a partial cross-sectional view of the slide processing apparatus of Figure 17 with the processing fluid collected at a lower end of a gap.
  • Figure 20 is a partial cross-sectional view of the slide processing apparatus of Figure 17 after a bolus of fluid is accumulated at the lower end of the gap.
  • Figure 21 is a side elevational view of a slide processing apparatus having a conformable opposable substrate.
  • Figure 22 is a side elevational view of the slide processing apparatus with processing fluid accumulated proximate to a waste remover.
  • Figure 23 is an isometric view of a slide processing apparatus, in accordance with one embodiment.
  • Figure 24 is another isometric view of the slide processing apparatus of Figure 23.
  • Figure 25 is a side elevational view of the slide processing apparatus of Figure 23.
  • Figure 26 is a cross-sectional view of the slide processing apparatus taken along a line 26-26 of Figure 23.
  • Figure 27 is an isometric view of a porous member spaced apart from a holder base.
  • Figure 28 is an isometric view of a holder base, in accordance with one embodiment.
  • Figure 29 is a bottom view of the holder base of Figure 28.
  • Figure 30 is an isometric exploded view of a slide handler, in accordance with one embodiment.
  • Figure 31 is an isometric view of a slide processing apparatus, in accordance with one embodiment.
  • Figure 32 is a side elevational view of the slide processing apparatus of Figure 31 .
  • Figure 33 is an isometric view of a substrate holder, in accordance with one embodiment.
  • Figure 34 is a detailed isometric view of a portion of the substrate holder of Figure 33.
  • Figure 35 is an isometric view of the substrate holder of Figure 33 holding a substrate and a slide on the substrate.
  • Figure 36 is a side elevational view of a registration feature of the substrate holder of Figure 33.
  • Figure 37 shows an automated processing system.
  • Figure 38 is an isometric view of an automated processing system having a staining system with an array of slide processing apparatuses, in accordance with one embodiment.
  • Figure 39 is an isometric view of a manifold system, in accordance with one embodiment.
  • Figure 1 shows a first substrate 80, a second substrate 82, and a substance 86 between the first and second substrates 80, 82.
  • the first and second substrates 80, 82 can be moved with respect to one another to manage the substance 86.
  • Managing the substance 86 may include spreading the substance 86 along an upper surface 90 of the substrate 80, moving a bolus of the substance 86 along the upper surface 90, or otherwise manipulating the substance 86 to process a sample 88 on the upper surface 90.
  • the first and second substrates 80, 82 can perform protocols using optimized liquid volumes to minimize or avoid problems with excessive volume consumption, including high processing costs.
  • a variable height gap 91 formed by the first and second substrates 80, 82 enables variable volume processing.
  • optimized volumes of liquids can be used for processing to increase efficiency and reduced cost as compared to fixed volume processing (i.e., processing that only uses a constant volume of liquid for each liquid application).
  • the reduction of costs may be based on the reduction of the consumed liquid volumes, as well as the reduction of system costs by reducing or avoiding relatively high costs associated with high liquid volume consumption, including manufacturing costs, packaging costs,
  • Dispensing of excessive liquid volumes may also lead to malfunctions (e.g., clogging, leaking, or the like) of fluidic components and may require frequent recalibration of equipment.
  • the gap 91 can accommodate a wide range of liquid volumes, even without moving the substrates 80, 82.
  • the gap 91 can accommodate liquid volumes in a range of about 10 microliters to about 200 microliters.
  • the height profile of the gap 91 can also be varied based on the liquid volume to be utilized.
  • the size of the gap 91 can be increased to avoid overfilling. Overfilling occurs when the volume of dispensed liquid is greater than the volume of the gap 91 (e.g., the volume between the first and second substrates 80, 82). Overfilling can lead to unwanted conditions, including sagging of liquid and ultimately fluid draining, especially if the substrates 80, 82 are at an inclined or upright orientation.
  • FIG. 1 shows under-filled regions 94a, 94b (collectively "94") of the gap 91 that can be filled with the liquid 86 by reducing the height of the gap 91 , changing the height profile, and/or by adding liquid.
  • 94 under-filled regions 94a, 94b (collectively "94") of the gap 91 that can be filled with the liquid 86 by reducing the height of the gap 91 , changing the height profile, and/or by adding liquid.
  • a significant volume of liquid can be conveniently added to the gap 91 without overfilling.
  • Processing protocols may require different liquid volumes in order to meet various processing criteria (e.g., chemical requirements, uptake
  • the substrates 80, 82 can be used to perform a processing protocol, including protocols involving applying different volumes of liquid. If the sample 88 is a paraffin embedded specimen, a relatively small volume of de-waxing solution (e.g., 12 microliters of xylene) can be delivered into the gap 91 .
  • the substrate 82 can apply the liquid to the sample 88.
  • the substrate 82 can be rolled (e.g., rolled along an imaginary plane spaced apart from the upper surface 90, rolled along the upper surface 90, or the like) or otherwise manipulated (e.g., rotated, translated, or both). After dewaxing, a relatively large volume of reagent can be delivered into the gap 91 .
  • a volume of about 120 microliters of stain can be delivered into the gap 91 .
  • the gap 91 has a minimum holding capacity of about 5 microliters (shown in solid line in Figure 1 ) and a maximum holding capacity of about 200 microliters (shown in dashed line in Figure 1 ).
  • the minimum holding capacity is the smallest volume of liquid that can be contained in the gap 91 and effectively applied to the sample 88.
  • the maximum holding capacity is the largest volume of liquid that can be contained in the gap 91 without overfilling.
  • the variable height gap 91 can accommodate a wider range of liquid volumes as compared to a uniform height gap because the narrowed region of the gap 91 can accommodate a small liquid volume, while the widened regions 94 of the gap 91 can accommodate large liquid volumes.
  • the widened regions 94 of the gap 91 can be conveniently accessed and filled.
  • At least a section of the substrate 82 can have a radius of curvature in a range of about 0.25 inch to about 20 inches.
  • the illustrated substrate 82 has a generally uniform radius of curvature. Other dimensions and curvatures are possible, if needed or desired. To adjust the holding capacity, the dimensions of the gap 91 can be selectively increased or decreased. This provides processing flexibility and efficiency.
  • the second substrate 82 can move the liquid 86 via capillary action.
  • the gap 91 is a capillary gap that can be maintained regardless of the presence or absence of liquid.
  • One portion of the capillary gap 91 can be narrower and have greater capillarity than a different portion of the capillary gap 91 .
  • the liquid 86 tends to flow to the narrowed portions of the gap 91 .
  • Figure 1 shows the entire substrate 82 spaced apart from the substrate 80 to avoid unwanted movement of the liquid 86. If the substrate 82 physically contacts the upper surface 90, the liquid 86 may tend to flow along the interface between the substrate 82 and the upper surface 90. Even though the entire substrate 82 is spaced apart from the substrate 80, the substrates 80, 82 can effectively enchamber the liquid 86. A minimum distance between the substrates 80, 82 can be maintained to provide the desired enchambering.
  • the substrates 80, 82 can physically contact one another for a period of processing or throughout the entire processing period.
  • the upper surface 90 is physically contacted to position the substrate 82.
  • the substrate 82 can include one or more discrete gapping elements.
  • the gapping elements can include outwardly protruding dimples that face the substrate 80 to keep the surface 92 of the substrate 82 spaced apart from the surface 90 of the substrate 80 so as to define the boundary of a processing area.
  • Gapping elements can also include, without limitation, one or more positioners, rails, spacers, or other structural features for serving as spacers.
  • the substrate 82 includes one or more rails ⁇ e.g., straight rails, arcuate rails, or the like) that bear against the upper surface 90.
  • the gapping elements may be separate components positionable between the substrates 80, 82, or at any other suitable location.
  • a first end 96 of the substrate 82 in Figure 1 can be moved towards the substrate 80 until the liquid 86 is in the position shown in Figure 2.
  • the liquid 86 can also be moved to an opposing second end 98 of the substrate 82 by narrowing the portion of the gap 91 formed by the second end 98, as shown in Figure 3. In this manner, the ends 96, 98 can be alternately lowered and raised to move the liquid 86.
  • Figure 4 shows the capillary gap 91 having a generally uniform height such that the liquid 86 fills a substantial volume of the gap 91.
  • the volume of the gap 91 is the volume directly between the first and second substrates 80, 82.
  • the range of holding capacities of the gap 91 of Figure 4 is narrower than the range of holding capacities of the variable height gap 91 in Figure 1 .
  • An excess or deficiency of about 1 -10 microliters may result in overfilling or under-filling.
  • the difference between the minimum holding capacity and the maximum holding capacity of the variable height gap 91 of Figure 1 can be about 25 microliters, 50 microliters, 100 microliters, or 150 microliters, or ranges encompassing such liquid volumes.
  • Evaporative losses can be controlled using the first and second substrates 80, 82.
  • the substrate 82 of Figure 1 in the curved configuration can expose a relatively large surface area of the liquid 86 to the surrounding environment.
  • the radius of curvature of the substrate 82 may be increased to reduce the surface area of the liquid 86 exposed to surrounding ambient air.
  • the second substrate 82 is a coverslip in a flat configuration to provide enhanced enchambering effect to appreciably mitigate or substantially eliminate evaporative losses.
  • the term "substrate” is a broad term and includes, but is not limited to, a slide, a coverslip, a strip of material, a plate, a carrier capable of carrying one or more samples, or the like. Substrates can be substantially rigid, semi-compliant, and/or compliant.
  • the substrate 80 is a microscope slide.
  • the substrate 82 is a compliant spooled film made, in whole or in part, of a transparent material, a non- transparent material, or both.
  • the second substrate 82 is a transparent coverslip.
  • a substrate can also be part of another component.
  • a platen or holder can have an outer surface that forms a substrate. The dimensions, properties (including mechanical properties, chemical properties, surface properties, and/or optical properties), and configurations of the substrates can be selected based on the processing protocol and subsequent analyses techniques to be performed.
  • a substrate is a substantially flat substrate.
  • substantially flat substrate refers, without limitation, to any object having at least one substantially flat surface, but more typically to any object having two substantially flat surfaces on opposite sides of the object, and even more typically to any object having opposed substantially flat surfaces, which opposed surfaces are generally equal in size but larger than any other surfaces on the object.
  • a substantially flat substrate can be formed of any suitable material, including glass, silicon, a semiconductor material, metal, combinations thereof, or the like.
  • Non- limiting examples of substantially flat substrates include slides (both 1 inch x 3 inch microscope slides and 25 mm x 75 mm microscope slides), SELDI and MALDI chips, silicon wafers, or other generally planar objects with at least one substantially flat surface.
  • the substrate 82 can assume a wide range of configurations.
  • Figures 1-3 show the substrate 82 in a simple arc configuration.
  • Simple arcs include arcs having a generally uniform curvature. The radius of curvature can be about 0.5 inch, 5 inches, 20 inches, 25 inches, or ranges encompassing such radii of curvature. Other radii are also possible.
  • the substrate 82 can assume a complex arc configuration or a compound arc configuration. If the substrate 82 is in the complex arc configuration, at least a portion of the substrate 82 has a varying curvature. If the substrate 82 is in a compound arc configuration, a portion of the substrate 82 can be a simple arc and another portion of the substrate 82 can be a complex arc.
  • FIG 5 shows a slide processing apparatus 100 including a substrate holder 1 10 and a waste remover 130.
  • the substrate holder 110 carries an opposable substrate 140 used to apply a processing liquid 160 to a specimen carried by a microscope slide 120.
  • the substrate 140 can mix the processing liquid, spread the processing liquid, enhance liquid uptake, control evaporation, or otherwise manage the processing liquid.
  • the illustrated substrate holder 1 10 engages a back face 141 of the substrate 140 such that a front face 200 (see Figure 6) of the substrate 140 faces the slide 120.
  • Processing liquids can be efficiently applied to biological samples to minimize or limit the cost of processing liquid(s) and to minimize and limit the amount of waste liquid produced.
  • the substrate 140 can be manipulated ⁇ e.g., translated, rotated, rolled, vibrated, or combinations thereof) to move the liquid 160 along the slide 120 using capillary action.
  • Capillary action can also be used to accumulate the liquid 160 at a desired location.
  • Forces, such as gravity, capillary forces, a pressure change ⁇ e.g., a reduced pressure such as a vacuum) in a gap 170, or combinations thereof can be used to remove the liquid 160.
  • Figure 5 shows the liquid 160 accumulated at an end 142 of the substrate 140 such that the waste remover 130 can aspirate the liquid 160.
  • Figures 6 and 7 show a bolus of liquid 160 at an end 166 of the gap 170.
  • the waste remover 130 applies a vacuum
  • the liquid 160 is drawn out of the gap 170 and into an inlet 185 of the waste remover 130.
  • the inlet 185 (illustrated in partial cross-section) is spaced apart from the gap 170 to avoid disruption of the surface tension of the liquid 160.
  • the waste remover 130 can remove waste liquids with a high level of precision and repeatability without unwanted wicking or without contacting a sample on the slide 120. This contactless removal of the liquid 160 can thus avoid problems associated with wicking away liquids.
  • the gap 170 can also be at an incline to facilitate fluid removal.
  • the end 166 of the gap 170 can be widened to allow the liquid 160 to pass out of the gap 170.
  • the end 166 is widened at least about 0.005 inch, 0.01 inch, 0.015, or 0.02 inch, or ranges encompassing such distances.
  • the height of the gap 170 at the end 166 is about 0.015 inch when the fluid 160 is removed.
  • substantially no residual liquid 160 remains in the gap 170 after the vacuum has been applied for a sufficient length of time.
  • Appropriate surface finishes ⁇ e.g., surface smoothness) and surface energy (e.g., the energy determined by the surface chemistry of the substrates 80, 82) can be selected to enhance the tendency of the liquid 160 to flow smoothly and completely from the gap 170. A higher level of smoothness and a lower surface energy will favor migration along the gap 170, whereas more surface finishes ⁇ e.g., surface smoothness
  • surface energy e.g., the energy determined by the surface chemistry of the substrates 80, 82
  • Figure 9 shows the emptied gap 170.
  • the surfaces 200, 210 can be made of a hydrophobic material and can have smooth finishes to facilitate the movement of the liquid 160.
  • the slide processing apparatus 100 can perform different tissue preparation processes and mounting processes.
  • Tissue preparation processes can include, without limitation, deparaffinizing a specimen, conditioning a specimen, staining a specimen, performing antigen retrieval, performing immunohistochemistry (IHC), and/or performing in situ hybridization (ISH), as well as other processes for preparing specimens for microscopy, microanalyses, mass spectromethc methods, or the like.
  • the specimen is a sample embedded in paraffin
  • the sample can be deparaffinized using appropriate deparaffinizing fluid(s).
  • the waste remover 130 removes the deparaffinizing fluid(s)
  • any number of reagents can be successively applied to the specimen.
  • the slide 120 can then be coverslipped with the substrate 140, or another coverslip.
  • Mounting processes include, without limitation, wet mounting slides, permanently mounting slides, or otherwise covering the sample.
  • Samples can be processed with a wide range of substances, such as reagents, probes, rinses, and/or conditioners.
  • the substances can be fluids (e.g., gases, liquids, or gas/liquid mixtures), or the like.
  • the fluids can be solvents (e.g., polar solvents, non-polar solvents, etc.), solutions (e.g., aqueous solutions or other types of solutions), or the like.
  • Reagents include, without limitation, stains, wetting agents, antibodies (e.g., monoclonal antibodies, polyclonal antibodies, etc.), antigen recovery fluids (e.g., aqueous- or non-aqueous-based antigen retrieval solutions, antigen recovery buffers, etc.), or the like.
  • Stains include, without limitation, dyes, hematoxylin stains, eosin stains, conjugates of antibodies or nucleic acids with detectable labels such as haptens, enzymes or fluorescent moieties, or other types of substances for imparting color and/or for enhancing contrast.
  • processing fluids in the form of reagents are applied to the samples.
  • concentrated liquids can be utilized. For example,
  • concentrated reagents can be uniformly applied over samples with large surface areas to reduce processing costs and waste.
  • a thin reagent film can be kept in contact with the sample to ensure enhanced uptake. Excessive volumes of reagents can be conveniently removed in a controlled manner.
  • the slide 120 is a generally flat transparent substrate capable of carrying a specimen for examination using equipment, such as optical equipment, e.g., a microscope or other optical device.
  • the slide 120 may be a generally rectangular piece of transparent material having a front face 210 for supporting specimens.
  • the slide 120 has a length of about 3 inches (75 mm) and a width of about 1 inch (25 mm) and, in certain
  • the slide 120 may include a label, such as a barcode.
  • the slide 120 has a length of about 75 mm, a width of about 25 mm, and a thickness of about 1 mm.
  • the microscope slide 120 can be in the form of a standard microscope slide made of glass or other transparent material.
  • the slide 120 can include a machine-readable code (such as a one- or multi-dimensional barcode or infoglyph, an RFID tag, a Bragg-diffraction grating, a magnetic stripe or a nanobarcode) with coded instructions that specify the type, sequence, and timing of the liquid(s) delivered for treatment of a particular specimen.
  • an actuation assembly 180 of the substrate holder 1 10 includes actuators 182, 184 that may be selectively extended and retracted to move the substrate 140.
  • the actuation assembly 180 can include, without limitation, one or more drives (e.g., linear drives, reciprocating drives, or the like), motors (e.g., stepper motors, drive motors, or the like), solenoids, piston assemblies, gear trains, combinations thereof, or other electronically, mechanically, hydraulically, or pneumatically driven components capable of moving the substrate 140.
  • the actuators 182, 184 and substrate 140 can be separate components.
  • the actuators 182, 184 can include chucks or other devices suitable for releasably holding the substrate 140.
  • the actuators 182, 184 can be permanently connected to the substrate 140.
  • a line 190 of Figure 6 extends from the actuation assembly 180 to the substrate 140.
  • the line 190 can be a vacuum line to apply a vacuum to hold the coverslip against the chuck.
  • the line 190 can be used to deliver fluid onto the slide 120.
  • Pressurized liquid can pass through the vacuum line 190 and through an aperture (not shown) in the substrate 140 (e.g., a unitary substrate physically connected to the actuators 182, 184) to dispense the fluid onto a front face 210 of the slide 120.
  • the illustrated dispensed liquid 160 is trapped between the lower surface 200 of the substrate 140 and the upper surface 210 of the slide 120.
  • the substrate 140 overlays most of or substantially the entire sample carried on the slide 120.
  • the substrate 140 has a shape generally similar to the shape of the mounting region of the slide 120. If the slide 120 is a standard microscope slide, the substrate 140 can have a length in a range of about 0.5 inch (13 mm) to about 3 inches (76 mm), a width in a range of about 0.5 inch (13 mm) to about 1 inch (25.5 mm), and a thickness in a range of about 0.02 inch (0.5 mm) to about 0.08 inch (2 mm).
  • the substrate 140 is a standard coverslip with a length of about 50 mm, a width of about 24 mm, and a thickness of about 0.2 mm. Other dimensions are also possible, if needed or desired.
  • the substrate 140 can have a generally polygonal shape (e.g., square or rectangular), elliptical shape, or circular shape. The shape of the substrate 140 can be selected based on the shape and dimensions of the slide 120, as well as the shape and dimensions of the sample.
  • the substrate 140 can be made, in whole or in part, of one or more polymers, plastics, composites, glass, combinations thereof, or other suitable materials that may be generally rigid, semi-rigid, or compliant.
  • the substrate 140 can be a rigid glass plate.
  • a flexible substrate 140 can be made of one or more polymers, such as polyester, polyethylene terephthalate,
  • the composition of the substrate 140 can be selected based on desired characteristics, including, without limitation, surface energy, flexibility, wettability, chemical compatibility, or the like.
  • the slide 120 and substrate 140 are made of a hydrophobic material to ensure sufficient containment of the liquid 160.
  • the waste remover 130 includes a pressurization device 220 and a receiving line 230 extending from the pressurization device 220.
  • the pressurization device 220 can draw the liquid 160 into the receiving line 230.
  • the pressurization device 220 may include, without limitation, one or more pumps, vacuum devices, or other types of devices capable of pressurizing fluids or drawing a vacuum, or both.
  • the pressurization device 220 can also include one or more waste reservoirs and/or can be connected to a separate waste reservoir. Waste can be delivered to the waste reservoir(s) for storage until subsequent disposal.
  • a disposal system is incorporated into the pressurization device 220.
  • waste received by the waste remover 130 is routed to an auxiliary disposal system. Waste can be conveniently disposed of without exposing operators or technicians, as well as other slide processing equipment, to waste.
  • the inlet 185 is spaced apart from an opening 215 of the gap 170.
  • the liquid 160 is drawn past edges 233, 235 of the slide 120 and substrate 140, respectively, into the inlet 185.
  • Adhesion forces, capillary forces, surface tension, or the like can keep the liquid 160 within the gap 170.
  • the inlet 185 may be spaced a sufficient distance from the liquid 160 to prevent inadvertent physical contact with the liquid 160 but close enough to allow convenient aspiration of the bolus of liquid 160 using a relatively low vacuum.
  • the end 185 is separated from the opening 215 by a distance D of at least about 0.001 inch (0.025 mm), 0.003 inch (0.076 mm), 0.02 inch (0.5 mm), 0.05 inch (1 .3 mm), or ranges encompassing such distances to avoid disrupting the surface tension.
  • the substrate 140 is drawn towards to slide 120 as the liquid 160 is removed. As such, the bottom surface 200 of the substrate 140 can be brought in physical contact with the upper surface 210 of the slide 120. In other embodiments, the substrates 80, 82 are kept spaced apart as the liquid 160 is removed.
  • the receiving line 230 can include, without limitation, one or more conduits, pipes, or other components through which fluid can flow.
  • the line 230 is a single lumen conduit. If the waste remover 130 delivers fluids onto the slide 120, the line 230 can be a multi-lumen conduit.
  • Liquids can be delivered through one lumen to the slide 120 and waste can be withdrawn from the slide 120 through another lumen.
  • the inlet 185 can include one or more openings 186, or other types of features, through which liquids can flow.
  • Figure 6 shows a height H of the gap 170.
  • the height H can be in a range of about 0.025 inches (0.635 mm) to about 0.1 inches (2.54 mm).
  • the nominal height H can be in a range of about 0.006 inch (0.15 mm) to about 0.01 inch (0.25 mm).
  • the height H can be in a range of about 0.001 inches (0.025 mm) to about 0.03 inches (0.76 mm).
  • Figure 7 shows the liquid 160 ready to be removed.
  • the height H can be increased or decreased to decrease or increase the forces needed to dislodge the bolus of liquid 160.
  • the height H can be about 0.003 inches (0.08 mm) when the pressurization device 220 draws a sufficient vacuum to overcome the adhesion forces between the liquid 160 and the slide 120/substrate 140.
  • Other heights H are also possible.
  • Figures 10-14 show one method of applying the liquid 160 to a sample 240 and removing the liquid 160. Generally, the liquid 160 is delivered between the slide 120 and the substrate 140. The substrate 140 is moved to apply the liquid 160 to the sample 240. The liquid 160 is then urged to a suitable location for removal. The liquid 160 is removed from the slide 120.
  • This method can be repeated to deliver any number of substances to perform a desired protocol, including protocols that involve successively applying different liquids (e.g., reagents) and/or applying the same liquid (e.g., rinse solutions, solvents, or the like) multiple times.
  • different liquids e.g., reagents
  • the same liquid e.g., rinse solutions, solvents, or the like
  • the sample 240 can be a biological specimen that includes one or more tissue sections, cytological preparations, micro-arrays (e.g., micro-arrays of DNA, protein, or the like), tissue arrays, cells, or other types of biological specimens.
  • a biological specimen can be any sample obtained from, derived from or containing any organism including a plant, an animal, a microbe or even a virus.
  • Particular examples of biological specimens include tissue sections, cytology samples, sweat, tears, urine, feces, semen, pre-ejaculate, nipple aspirates, pus, sputum, blood, serum, tissue arrays, and protein and nucleic acid arrays.
  • the illustrated sample 240 is a single tissue section, such as an embedded tissue section (e.g., a paraffin embedded tissue section), lying on the slide 120.
  • the liquid 160 can flow through the delivery line 190 to at least partially fill the gap 170.
  • the liquid 160 flows along the gap 170 via capillary action towards the periphery of the substrate 140.
  • Capillary action can include, without limitation, movement of the liquid 160 due to the phenomenon of the liquid 160 spontaneously creeping through the gap 170 due to adhesive forces, cohesive forces, and/or surface tension.
  • the volume of liquid 160 can be in a range of about 50 ⁇ l_ to about 700 ⁇ l_.
  • the liquid 160 flows along the gap 170 to coat most of or the entire specimen 240.
  • the nominal height of the gap 170 is equal to or less than about 0.003 inch (0.08 mm) and the volume of liquid 160 is about 100 ⁇ l_.
  • the substrate 140 can be actively moved, passively moved, or both.
  • the actuators 182, 184 can actively move the substrate 140 independent of the capillary forces.
  • capillary forces can cause movement of the substrate 140 towards the slide 120.
  • the liquid 160 can provide self-alignment and/or self-gapping. For self-alignment, the substrate 140 can be moved to a substantially parallel orientation with respect to the substrate 120 due to the forces provided by the liquid 160.
  • the substrates 80, 82 are parallel to within about 0.01 inch (0.25 mm), 0.005 inch (0.127 mm), 0.003 inch (0.07 mm), or 0.001 inch (0.03 mm).
  • parallelism of the substrates 80, 82 can be within about 0.003 inch (0.07 mm).
  • the substrate 140 lies in a plane that forms an angle that is less than about 10 degrees with an imaginary plane in which the slide 120 lies.
  • the liquid 160 can draw the substrate 140 towards the slide 120 to provide self-gapping. Once the forces are balanced, the substrate 140 can be generally static. In some embodiments, the forces can be balanced to avoid overfilling and under-filling. The liquid 160 thus moves the substrate 140 to a desired position without requiring operation of the actuators 182, 184.
  • the liquid 160 can provide a force less than about 50 gram-force, 25 gram-force, 10 gram-force, 1 gram-force, or ranges encompassing such forces for self-gapping.
  • the extendable members 182, 184 can allow substantially free movement of the substrate 140 in response to forces provided by the liquid 160.
  • the gap 170 can stabilize at a full fill state corresponding to when the gap 170 is completely filled with the liquid 160.
  • the extendable members 182, 184 provide substantially no resistance such that the substrate 140 floats on the liquid 160.
  • the capillary forces can equilibrate to form a thin layer of liquid 160 with a substantially uniform height.
  • the ends 142, 192 of the substrate 140 can be alternatingly raised and lowered to move the liquid 160 back and forth across the specimen 240 any number of times.
  • the other end 142 of the substrate 140 is moved away from the slide 120.
  • the liquid 160 is urged towards a narrowed region 247 of the gap 170 via capillary action as an opposing region 249 of the gap 170 widens.
  • the bolus of liquid 160 at the end 192 of the substrate 140 can be reapplied to the specimen 240 by raising the end 192.
  • the end 142 is moved downwardly and the end 192 is moved upwardly. As the region 249 narrows, the liquid 160
  • Figure 13 shows the liquid 160 accumulated at the narrowed region 249.
  • the waste remover 130 can aspirate the bolus of liquid 160.
  • the end 142 of the substrate 140 can be moved away from the slide 120.
  • the angled lower surface 200 in Figure 13 is rotated away from the slide 120, the liquid 160 is urged closer to the opening 215.
  • the substrate 140 can be moved to a generally parallel orientation to the slide 120 to keep the liquid 160 as close as possible to the waste remover.
  • the slide 120 can also be tilted. After the liquid 160 is accumulated, the substrate 140 can be moved away from the slide 120 to allow convenient removal of the bolus of liquid 160. For example, the height of the opening 215 can be increased until the bolus of fluid 160 passes through the opening 215. The minimal dimensions of the opening 215 for liquid removal can be determined empirically, if needed or desired.
  • the method of Figures 10-14 can be employed to accumulate liquid at a wide range of locations, including at the corners, sides, and/or ends of the substrate 140 and/or slide 120.
  • the position of the waste remover 130 can be selected based on the desired location of liquid accumulation.
  • a fluid dispenser can deliver a liquid into the empty gap 170.
  • Figure 14 shows a fluid dispenser 251 with an outlet 253 positioned to output a liquid onto the slide 120.
  • the fluid dispenser 251 can include, without limitation, one or more fluid containers, pumps, filters, valves, combinations thereof, or the like.
  • an adhesive in a liquid state can be delivered into the gap 170 after preparing the specimen 240.
  • the adhesive can include, without limitation, thermosetting materials, epoxy, or other curable or selectively hardening adhesives, as well as thermoplastics.
  • a thermosetting material can be a plastic that becomes permanently hardened when set. In some embodiments, the thermosetting material is cured using thermal energy, light energy, chemical activation or the like.
  • the adhesive comprises silicone, urethane resin, blends, or the like.
  • the adhesive can fixedly couple the substrate 140 to the slide 120 to form a permanent mount slide, thereby increasing the useful life of the slide, protecting the specimen 240 from environmental changes, and protecting against accidental movement of the coverslip.
  • the processing apparatus 100 can include one or more energy sources 270 (shown in dashed line in Figure 14) that output light energy.
  • the substrate 140 can be transparent or semi-transparent to allow propagation of the light to the adhesive.
  • a control system 272 can control the illumination sequence.
  • the adhesive can be cured in less than about 5 minutes, 60 seconds, 20 seconds, or 10 seconds. Other curing times are also possible, if needed or desired.
  • a coverslip can be deposited on the slide 120 without using any adhesive.
  • the substrate 140 is a coverslip used to form a wet mount slide.
  • the fluid dispenser 251 can output water for wet mounting.
  • Figure 15 shows a slide processing apparatus 300 capable of moving a microscope slide 304 and an opposing substrate 306 to different orientations, including a generally vertical orientation ( Figure 15), inclined orientation ( Figure 16), generally horizontal orientation, or the like.
  • the apparatus 300 includes a fluid dispenser 320 for dispensing a processing fluid and a waste remover 330 for removing waste.
  • the slide 304 can be at the inclined orientation or generally vertical orientation.
  • the apparatus 300 can be rotated about an axis of rotation 310. In some protocols, fluid is both dispensed and removed when the microscope slide 304 is in a generally vertical orientation.
  • the slide 304 can be at a generally horizontal orientation to help position (e.g., self-align, self-gap, etc.) the substrate 306.
  • the slide 304 and the substrate 306 can be rotated at different times during a particular protocol for increased processing flexibility.
  • the microscope slide 304 and substrate 306 are positioned at an inclined orientation (e.g., the slide 304 and substrate 306 can be at angle of inclination of about 10 degrees, 20 degrees, 45 degrees, 70 degrees, or 80 degrees).
  • Liquid can be delivered between the microscope slide 304 and the substrate 306.
  • the microscope slide 304 and substrate 306 can be moved from the inclined orientation to another orientation to move the liquid along the capillary gap.
  • the slide 304 and the substrate 306 can slope upwardly at an angle of inclination ⁇ within a range of about 10 degrees to about 80 degrees while the liquid is delivered.
  • the angle of inclination ⁇ is in a range of about 30 degrees to about 60 degrees, in a range of about 40 degrees to about 50 degrees, or other suitable ranges of angles for facilitating liquid delivery.
  • the microscope slide 304 and substrate 306 can be rotated about the axis of rotation 310.
  • the substrate 306 and slide 304 can be inverted any number of times.
  • the slide processing apparatus 300 of Figure 16 can be rotated about the axis of rotation 310 at least about 45 degrees, 80 degrees, 90 degrees, or 180 degrees after the liquid is dispensed.
  • the fluid dispenser 320 can include, without limitation, valves, thermal elements (e.g., heaters, coolers, etc.), controllers, or the like and can output a wide range of substances, including mounting media, reagents, or other processing fluids.
  • the fluid dispenser 320 includes one or more thermo-electric elements 393 for controlling the temperature of the substance outputted from an outlet 395. If the dispensed substance is an adhesive, the elements 393 can heat the adhesive to adjust the viscosity, spreadability, or other characteristics of the adhesive.
  • the fluid dispenser 320 is in the form of a heated pipette.
  • a frame 349 carries a slide handler 350 and a substrate handler 380.
  • the frame 349 can be a generally rigid structure capable of supporting both the slide handler 350 and the substrate handler 380.
  • the frame 349 is a generally U-shaped unitary member with ends connected to the slide handler 350 and the substrate handler 380.
  • the frame 349 has a multi-piece
  • the slide handler 350 includes a holder 360 for carrying the microscope slide 304 and a slide positioning device 370 for moving the slide 304 towards or away from the substrate 306, as indicated by the arrows 372, 374.
  • the holder 360 can be a mechanical chuck with one or more clamps, adhesive layers, mechanical fasteners, or the like capable of selectively holding and releasing the slide 304.
  • the holder 360 is a vacuum chuck or electrostatic chuck. Other types of holders can also be used.
  • a substrate handler 380 includes a holder 382 configured to carry the substrate 306. Additionally or alternatively, the holder 382 can release the coverslip 306 using a positive pressure. If the substrate 306 is reusable, the holder 382 may hold the substrate 306 for any number of processing protocols. For example, the substrate 306 can be used to perform multiple staining protocols. To perform a different protocol, the substrate 306 can be removed and discarded to prevent carryover. In other embodiments, the substrate 306 is replaced during a single protocol. For example, different substrates can be used to apply different reagents to a single specimen.
  • the substrate 306 can be a length of a strip of material.
  • the slide processing apparatuses 300 can be used in combination with the strips, systems, components, and features disclosed in U.S. Patent Application No. 1 1/187,183 (Publication No. 2006/0019302), which is incorporated by reference in its entirety.
  • the substrate 306 can be a strip of material that is moved between the slide 304 and the holder 382.
  • the holder 360 includes thermo-electric elements 390a, 390b
  • thermo-electrical elements 392a, 392b, 392c (illustrated in dashed line) adapted to convert electrical energy to thermal energy.
  • the thermo-electric elements are discussed in connection with the holder 360, but the description of the thermo-electric elements 390a, 390b (collectively "390") applies equally to the thermo-electrical elements 392a, 392b, 392c, unless indicated otherwise.
  • thermo-electric elements 390 can support different protocols that require thermal cycling, even rapid thermal cycling for ISH, IHC, or the like. When the elements 390 generate heat, heat is transferred to the slide 304. Heat is ultimately transferred from the slide 304 to the specimen(s) and processing liquid. The amount of electrical energy delivered to the elements 390 can be increased or decreased to increase or decrease the temperature of the
  • the elements 390 can be resistive heating elements.
  • resistive heating elements e.g., plate resistive heaters, coil resistive heaters, strip heaters, or the like
  • Other types of thermal elements such as cooling elements, heating/cooling elements, or the like, can be utilized.
  • cooling element is a broad term that includes, without limitation, one or more elements capable of actively absorbing heat so as to effectively cool at least a portion of the slide 304.
  • a cooling element can be a cooling tube or channel through which a chilled fluid flows.
  • the holder 360 includes heating elements for producing heat during a heating period and cooling elements for absorbing heat during a cooling period.
  • the elements 390 are heating/cooling elements, such as Peltier devices.
  • Peltier devices may be solid state components which become hot on one side and cool on an opposing side, depending on a direction of current passed therethrough. By simply selecting the direction of current, the Peltier device can be employed to heat the slide 304 for a desired length of time. By switching the direction of the current, the elements 390 cool the slide 304.
  • the heating/cooling elements 390 can be in the form of channels through which a working fluid flows. Heated fluid can be passed through the channels for a heating period, and a chilled fluid can be passed through the channels for a cooling period. The position, number, and type of heating/cooling elements 390 can be selected based on the desired temperature profile of the holder 360.
  • the elements 390 can also be light sources capable of converting electrical energy to optical energy.
  • the elements 309 can include, without limitation, one or more light emitting diodes, optical filters, optical fibers, scattering mediums, or other optical components that can be used to obtain a desired light distribution. Light can be distributed evenly or unevenly. If UV curable adhesive is used to coverslip the slide 304, the light sources 390 can be UV light sources.
  • the elements can be other types of light sources used before, during, or after processing of the specimen and/or during coverslipping.
  • Figure 17 shows the substrate handler 380 with an actuation assembly that includes magnets 400a, 400b (collectively "400") and elements 410a, 410b (collectively "410") that move away from and towards the respective electro-magnets 400.
  • the substrate holder 380 can have a different modes of operation, including an active mode (e.g., an actuation mode) and a passive mode (e.g., a self-alignment/self-gapping mode).
  • the active mode e.g., the electromagnets 400 and elements 410 cooperate to move a holder 482.
  • the passive mode the electro-magnets 400 can be OFF to allow substantially free movement of the holder 482, thereby allowing movement of the substrate 306 due to forces provided by a liquid 488.
  • a biasing mechanism 420 in the form of a flexure extends across a housing 430.
  • a retainer 440 of the holder 482 is coupled to a central section of the flexure 420.
  • the elements 410 extend away from the holder 482, past the flexure 420, and towards the respective magnets 400.
  • one or both of the magnets 400 can be energized to attract the respective elements 410.
  • the flexure 420 deforms to allow stops 480a, 480b (collectively "480") to move against the respective magnets 400.
  • the magnets 400 are de-energized, the flexure 420 can bias the substrate 306 towards the slide 304. In this manner, the flexure 420 and the magnets 400 cooperate to controllably move the substrate 306 away from and towards the slide 304.
  • the elements 410 can be made, in whole or in part, of one or more ferromagnetic materials (e.g., iron, steel, stainless steel, combinations thereof, or the like) that are attracted to magnets.
  • ferromagnetic materials e.g., iron, steel, stainless steel, combinations thereof, or the like.
  • the 410 can be magnets (e.g., an electromagnet) and the elements 400 can be made of a ferromagnetic material.
  • Figure 17 shows the substrate 306 angled with respect to the slide
  • the opening 484 can have a width in a range of about 0.025 inch (0.64 mm) to about 0.03 inch (0.76 mm). In some embodiments, the opening 484 can have a width that is equal to or greater than about 0.5 inches (13 mm), 0.3 inch (7.6 mm), 0.1 inch (2.5 mm), 0.05 inch (1 .27 mm), or .01 inch (0.25 mm), or ranges encompassing such widths.
  • the flexure 420 biases a lower end 486 of the substrate 306 towards the slide 304.
  • the lower end 486 of the substrate 306 can be proximate to the microscope slide 304 to capture the liquid 488 that falls from the fluid dispenser 320.
  • the magnet 400a is de-energized, and the flexure 420 moves an upper end 492 of the substrate 306 towards the slide 304.
  • the fluid 388 moves upwardly along the gap 499 due to capillary action.
  • Figure 18 shows a thin layer of the liquid 488 after the liquid 488 has been spread along the substrate 306.
  • Capillary forces F c pulling the substrate 306 move the holder 482 from an unbiased position (shown in dashed line) to a biased position 427.
  • the capillary forces can equilibrate to ensure that the substrate 306 is generally parallel to the slide 304.
  • the flexure 420 can allow substantially free movement of the substrate 306 in response to capillary forces provided by the liquid 488.
  • the suspended arrangement of the holder 482 allows the substrate 306 to effectively float on the liquid 488. Forces can be balanced to control the liquid 488 without having to utilize high precision positioning/alignment control devices that may be prone to errors, require recalibration, and other complicated moving components that are subject to various problems.
  • the flexure 420 can be made, in whole or in part, of one or more metals ⁇ e.g., spring steel, aluminum, or the like), polymers, composites, rubber, combinations thereof.
  • the flexure 420 can include, without limitation, one or more beam elements, flexible elongate members, springs (for example, leaf springs), or the like.
  • the flexure 420 has a length of about 3 inches (7.6 cm) and a diameter or thickness of about 0.002 inch (0.05 mm).
  • Such a flexure can be made of steel or other highly deformable material.
  • Figure 19 shows the element 410a contacting the magnet 400a to form a narrowed region 498 of the gap 499 at the lower end 486 of the slide 304.
  • the magnet 400b is de-energized.
  • the flexure 420 keeps the lower end 486 of the substrate 306 against or close to the slide 304. After the liquid 488 has moved downwardly towards the lower end 486, the magnet 400b is energized to move the lower end 486 away from the slide 304.
  • Figure 20 shows the lower end 486 spaced from the slide 304.
  • a bolus of liquid 488 at the lower end of the gap 499 is ready to be withdrawn by a waste remover 496.
  • an inlet 501 of the fluid remover 496 is fixed relative to the slide 304. This prevents or minimizes inadvertent contact between the fluid remover 496 and the slide 304 to prevent unwanted wicking or migration of the liquid 488.
  • the inlet 501 is movable relative to the slide 304.
  • the end 501 may be moved towards the accumulated liquid 488 or away from the accumulated liquid 488 based on, for example, the height of the gap 499, the properties of the liquid 488, surface properties of the substrate 306 and/or the slide 304, or the like.
  • Airflow or other fluid can also be used to facilitate removal of the processing fluid.
  • the fluid dispenser 320 of Figure 20 can output an airflow that passes through the gap 499 to push the liquid 488 towards the waste remover 496.
  • a conformable substrate 500 is capable of assuming a wide range of different configurations.
  • Figure 22 shows the substrate 500 in a generally flat configuration and angled with respect to a microscope slide 504 to collect a liquid 502 proximate to a fluid remover 520.
  • the substrate 500 can be made, in whole or in part, of a flexible polymer, plastic, composite material, membrane, combinations thereof, or other highly compliant material that can be repeatedly deformed.
  • liquid 502 can fill a narrowed region 510 of a gap.
  • Actuators in the form of extendable members 514a, 514b, 514c can be operated to move the substrate 500 to a generally flat configuration to form a generally uniform layer of liquid 502 extending across a specimen (not shown). After the liquid 502 is applied to the specimen, the liquid 502 can be collected near an inlet 519 of a waste remover 520.
  • the conformable substrate 500 can include gapping elements 515a, 515b, 515c, 515d, 515e, 515f, 515g (collectively "515").
  • the elements 515 space a lower surface 516 of the substrate 500 from the substrate 504.
  • the heights of the elements 515 can be greater than the thickness of the sample to prevent damage to the sample due to, for example, abrasion, compression, or the like. In this manner, a minimum gap height can be maintained.
  • FIGS 23-25 show a substrate holder 600 that includes an actuation assembly 609.
  • the actuation assembly 609 includes a pair of actuation mechanisms 610a, 610b (collectively "610") in the form of linear actuators adapted to move a holder 612.
  • the holder 612 includes a holder base 620 carrying a permeable member 622.
  • a frame 631 is rotatable about an axis of rotation 633 and carries the substrate holder 600.
  • the actuation mechanisms 610a, 610b can be generally similar to each other, and accordingly, the description of one actuation mechanism applies equally to the other, unless clearly indicated otherwise.
  • the actuation mechanism 610a of Figure 23 includes a drive unit 640, an extendable member 642, and a coupler 646 carried by the extendable member 642.
  • the coupler 646 has a narrowed region 648 and an enlarged tip 649. When the narrowed region 648 is positioned within a slot 650 of the holder base 620, the actuation mechanism 610a can push the holder 612 downwardly or pull the holder 612 upwardly. To replace the holder 612, the coupler 646 can be slid out of the slot 650.
  • Figure 26 shows a retainer 660 of the holder 612 including a biasing member 670 for offsetting the weight of the holder 612 to keep the holder 612 at a general neutral position.
  • the biasing member 670 can include, without limitation, one or more springs that can provide desired resistance to movement of the holder 612.
  • An upper end 672 of the spring 670 is coupled to a housing 674 and a lower end 680 of the spring 670 is coupled to the retainer 660.
  • a passageway 690 through the holder body 620 provides communication between the permeable member 622 and a pressurization line 654.
  • a vacuum can be drawn through the line 654 and the passageway 690 to hold the permeable member 622 against the holder body 620, as well as a substrate against a substrate contact face 623 of the member 622.
  • the holder base 620 includes a recessed region 700 sized and configured to receive the permeable member 622.
  • An opening 702 of the passageway 690 provides fluid communication between the recessed region 700 and the pressurization line 654.
  • a sidewall 710 can closely surround the periphery of the permeable member 622 to limit, minimize, or substantially prevent side-to-side movement of the permeable member 622.
  • the holder base 620 can have any number of passageways, channels, or other features. As shown in Figure 26, a gap 730 is between a lower surface 740 of the holder base 620 and an upper surface of the permeable member 622. Spacers 741 can keep the permeable member 622 spaced apart from the lower surface 740.
  • the permeable member 622 can be a porous member made of ceramic, metal, composite materials, combinations thereof, or other suitable material through which a vacuum can be drawn.
  • the porosity, composition, and thickness of the permeable member 622 can be selected based on the desired rate of airflow therethrough.
  • the permeable member 622 can be conveniently replaced with another permeable member, or another type of component.
  • a permeable member with a curved substrate contact face can be installed in order to bend and hold a substrate.
  • the substrate contact face of the permeable member can be concave, convex, or have any other suitable configuration.
  • Figures 28 and 29 show one embodiment of a holder base 620 including an array of interconnected channels 760 and a lower surface 740.
  • the channels 760 provide communication between the opening 762 and the permeable member or substrate overlying the surface 740.
  • the channels 760 of Figures 29 include an outer channel 780 extending about the periphery of the holder base 620 and channels 782, 784, 786, 788 extending from the opening 762 to the outer channel 780.
  • the pattern and configuration of the channels 760 can be selected based on the characteristics of the desired airflow.
  • sensors 770a, 770b can detect a measurable parameter and can send a signal indicative of that measured parameter.
  • the sensors 770 are position sensors capable of determining the position of the holder 612.
  • the holder base 620 has outwardly extending tabs 772a, 772b (see Figure 28) positioned underneath the sensors 770a, 770b, respectively.
  • the sensors 770 can be temperature sensors, motion sensors, chemical sensors, or the like.
  • Figures 23 and 24 show a flexure 800 that couples the holder 612 to a main body 802.
  • the flexure 800 is configured to provide motion of the holder 612 along the X-axis, Y-axis, and/or Z-axis.
  • the holder 612 can be moved from side to side along the X-axis and/or longitudinally along the Z-axis.
  • the flexure 800 also allows the holder 612 to move away from and towards the slide 624 along the Y-axis.
  • the flexure 800 includes outwardly extending arms 804a, 804b, 804c (collectively "804") extending from the holder 612 to the two legs 808a, 808b, 808c (collectively "808") of the housing 802.
  • the arms 804 have a corrugated configuration and extend, contract, bend, or combinations thereof. Each end of the arms 804 is fixedly coupled to the main body 802, and a central region 805 of the flexure 800 is fixedly coupled to the holder 612.
  • the holder 612 can rotate about the X-axis, about the Y-axis, and/or about the Z-axis and/or can translate along the X-axis, the Y-axis, and/or the Z-axis.
  • the number of arms of the flexure can be selected based on the desired motion of the holder 612.
  • the flexure 800 can be made, in whole or in part, of one or more metals, polymers, plastics, rubbers, metals, cloth, or combinations thereof.
  • the flexure 800 has a unitary construction and is made of fluoropolymers.
  • the flexure 800 can be made of TEFLON®.
  • the illustrated flexure 800 is substantially parallel to the holder 612 to provide a relatively small biasing force to allow floating a substrate.
  • Figure 25 shows a slide handler 820 connected to the frame 631 (see Figure 23).
  • the slide handler 820 is capable of pneumatically holding a microscope slide 614 by applying a vacuum via a line 840.
  • the slide handler 820 includes a plenum body 850 and an outer body 852.
  • the temperature of the plenum body 850 can be controlled via one or more lines 824. Heated fluid, chilled fluid, or the like can be passed through the plenum body 850 via the line 824 to control processing temperatures.
  • the plenum body 850 can include one or more networks of passageways to provide a desired temperature distribution. In some embodiments, chilled fluid is delivered into the plenum body 850 via one line 824, and fluid flows out of the plenum body 850 via another line 824.
  • thermal elements can be incorporated into the plenum body 850 and/or the outer body 852.
  • a thermal element such as a heater, is positioned between the plenum body 850 and the outer body 852. The heater can heat the specimen. After processing, fluid can be circulated through the plenum body 850 to cool the specimen. In this manner, the temperature of the specimen can be controlled to provide enhanced control of chemical reactions.
  • Figure 30 shows the outer body 852 including a recessed region 854 configured to receive the plenum body 850.
  • the plenum body 850 can be coupled to the outer body 852 mechanically by one or more fasteners (e.g., clips, nut and bolt assemblies, or the like), adhesives, or the like.
  • fasteners e.g., clips, nut and bolt assemblies, or the like
  • channels 878a, 878b are fluidly coupled to the line 840 ( Figure 25) via a port 857.
  • the channels 878a, 878b can extend along a substantial portion of longitudinal length of the plenum body 850.
  • a transverse channel 882 extends between the channels 878a, 878b.
  • a through-hole 872 couples the channel 882 to the port
  • the slide handler 820 also includes registration features 844 for positioning a slide.
  • the registration features 844 of Figure 30 are in the form of stops that extend upwardly past a slide contact face 880 of the plenum body 850. When an end 847 of the slide 614 abuts the stops 844, the slide 614 is spaced apart from an end 844 of a waste remover 849, as shown in Figure 25.
  • 614 can remain generally stationary as waste is removed.
  • FIGs 31 and 32 show a slide processing apparatus 900 holding a slide 910 at an inclined orientation.
  • the processing apparatus 900 includes a flexure 920 with generally planar arms 922a, 922b (collectively "922").
  • Each of the planar arms 922 can be a flexible plate of compliant material (e.g., spring steel, stainless steel, aluminum, titanium, or the like). Actuation mechanisms
  • a rotator unit 950 can rotate a slide handler 926 about the X-axis.
  • the substrate holders disclosed herein can include a waste port
  • FIGS 33-35 show a substrate holder 960 that includes a curved surface 962 that can be overlaid with a substrate.
  • Vacuum channels 968a, 968b are positioned along the surface 962.
  • Figure 34 shows a substrate 970 (illustrated in dashed line) with an edge 972 positioned adjacent to a waste remover 980 in the form of an aspiration port. Waste can be drawn into an opening 982 of the aspiration port 980.
  • Figure 35 shows a slide 990 angled with respect to the substrate 970 to position liquid waste along the edge 972. The waste is then pulled into the aspiration port 980.
  • Processing fluid can be delivered into the capillary space on the opposite side of the aspiration port 980 via a fluid dispenser 987 ( Figure 35) or on the same side as the aspiration port 980. Movement of the liquid is controlled using capillary tension. The slide 990 can be rotated to move the liquid towards the aspiration port 980 and allow waste aspiration.
  • the aspiration port 980 can include one or more registration features.
  • Figure 36 shows a registration feature 1000 for positioning the substrate 970.
  • the edge 972 of the substrate 970 can abut a vertical sidewall 973 of the registration feature 1000 to align the substrate 970 with the surface 962.
  • the registration feature 1000 can be a notch, protrusion, or other alignment feature formed in the aspiration port 980. In other embodiments, registration features are positioned along the surface 962.
  • the substrate holder 960 is undercut to substantially prevent contact between the liquid and the substrate holder 960.
  • the substrate 970 extends outwardly past the edge of the substrate holder 960 such that the liquid can be aspirated without wicking or capillary action due to the liquid physically contacting the substrate holder 960.
  • the substrate 970 can extend outwardly past the periphery of the substrate holder 960.
  • Figure 37 shows a processing system 1 100 that includes a staining system 1 105, a fluid handling system 1 1 10, a slide system 1 1 16, and a substrate system 1 1 18.
  • the staining system 1 105 can include slide processing
  • the staining system 1105 can include an array of stations, each with a slide processing apparatuses for parallel processing.
  • the staining system 1 105 includes a bank of stationary slide processing apparatuses.
  • the staining apparatus 1 105 includes a movable carousel with slide processing apparatuses. Valve mechanisms, temperature control systems, or other systems or components can be
  • each station includes a fluid dispenser, a processing apparatus, and a manifold system.
  • the fluid handling system 1 110 can include containers for holding fluid.
  • the containers can be connected to the staining apparatus 1 105 by one or more fluid lines.
  • Fluids in the containers may be mounting media, reagents, probes, rinses, and/or conditioners, and may be in the form of a fluid (e.g., gases, liquids, or gas/liquid mixtures).
  • the fluids can be solvents ⁇ e.g., polar solvents, non-polar solvents, etc.), solutions (e.g., aqueous solutions or other types of solutions), or the like.
  • Substances from the containers can be used to perform different protocols, such as staining protocols (e.g., primary staining, special staining, IHC, ISH, or the like), antigen retrieval protocols, or the like.
  • the fluid handling system 1 1 10 can also include one or more pumps that supply the fluid.
  • the fluid handling system 1 110 may include fixed nozzles, pipette systems, or other types of fluid dispensers. Fixed nozzles are especially well suited to delivery H&E fluids, bulk advanced stain fluids, or the like. Pipette systems are especially well suited to output non-bulk advanced stain fluids.
  • the slide system 1 1 16 can provide slides carrying samples ready for processing.
  • the slide system 1 116 can include, without limitation, heaters or slide dryers (e.g., conductive dryers, convection dryers, ovens, etc.), as well as other types of components or devices used to prepare samples.
  • the slide system 11 16 can also include any number of racks, trays, cartridges, or other structures suitable for holding a desired number of slides.
  • One or more slide transporters e.g., conveyors, robotic arms, or delivery mechanisms
  • the substrate system 1 118 can include, without limitation, one or more racks, trays, cartridges, receptacles, or any other structures suitable for holding a desired number of substrates.
  • One or more substrate transporters can deliver substrates between components of the substrate system 1 1 18.
  • the processing system 1 100 further includes a control system 1120 that communicates with various components.
  • the control system 1 120 is communicatively coupled to the stain system 1 105 by a wired connection 1 122 and is communicatively coupled to the fluid handling system 11 10, slide handling system 1 1 16, and substrate handling system 1 1 18 by wire connections 1 124, 1 126, 1 128, respectively. Communication can also be accomplished through wireless connections (including wireless network connections) and/or optical connections.
  • the control system 1 120 can generally include, without limitation, one or more computers, central processing units, processing devices,
  • control system 1 120 includes, without limitation, one or more storage elements, such as volatile memory, non-volatile memory, read-only memory (ROM), random access memory (RAM), or the like.
  • the stored information can include optimization programs, tissue preparation programs, calibration programs, indexing programs, or other executable programs.
  • the control system 1 120 can execute optimization programs to optimize performance ⁇ e.g., reduce excess reagent consumption, reduce coverslipping time, increase productivity, improve processing consistency, or the like).
  • the processing may be optimized by determining, for example, an optimum schedule to increase processing speeds, to increase throughput (e.g., a number of slides processed in a length of time), or the like.
  • Such optimum schedule can be a schedule of preparing and delivering slides to the stain system 1 105.
  • the control system 1 120 determines loading sequences to reduce processing wait times.
  • the control system 1 120 can also be programmed such that loading of the pipettes or nozzles for the next specimen can start during processing of the currently loaded specimen. This saves time because fluids can be dispensed onto the next specimen as soon as the current specimen is removed from the slide handler.
  • the processing system 1 100 can include any number of the transporters.
  • Transporters can include, without limitation, one or more robotic handlers, X-Y-Z transport systems, conveyors, slide systems, combinations thereof, or other automated mechanisms capable carrying items between locations, if needed or desired.
  • Transporters can have end effectors to carry items.
  • End effectors may include, without limitation, grippers, suction devices, holders, or the like.
  • Sensors of the transporters can be temperature sensors, vacuum sensors, surface sensors, position sensors, or the like.
  • vacuum sensors in an end effector are capable of detecting substrate or slide integrity. The end effectors can load the slide processing apparatuses with both slides and substrates. After processing, the end effectors can retrieve the slides so that the process can be repeated.
  • Figure 38 shows one embodiment of the staining system 1 105 including a carousel of spaced apart processing apparatuses.
  • Transporter 1 140 includes linear slides, cartridges, and end effectors for transporting slides from the slide system 1 116 to the individual processing apparatuses.
  • the transporter 1 140 can also carry substrates from the substrate system 1 118 (illustrated as a container 1143 holding coverslips).
  • the slide system 1 1 16 includes a slide rack 1 144 for holding slides in an upright position. Ends of the slides can be conveniently gripped by an end effector of the transporter 1140 to pull the slides from the rack 1 144. The slides can be moved to a dryer or moved directly to processing apparatuses. A slide rack positioning device 1150 can position the rack 1 144 with respect to the transporter 1 140.
  • Figure 39 shows a manifold system 1160 of the staining apparatus
  • valves and sensors for waste disposal, coverslipping, slide vacuum, thermal processing (e.g., cooling and cooling purge), detecting coverslip and/or slide integrity, detecting waste evacuation, combinations thereof, or the like.
  • the manifold system 1 160 can include valves, controllers, fluid lines, or other components used to control and/or evaluate parameters of interest.
  • Each processing apparatus can be connected to a respective manifold system.
  • information, if any, on each slide may be read by a reader.
  • slides may have a code. Instructions that are on the code, or are referenced by the code, are used to determine the treatments to be performed on a given slide on one or more of the processing apparatuses.
  • the processing apparatuses can de-paraffinize paraffin-embedded tissue sections and perform staining protocols (such as a primary stain, an IHC stain, or an ISH stain).
  • staining protocols such as a primary stain, an IHC stain, or an ISH stain.
  • a slide transporter 1 171 of Figure 38 (illustrated as an X-Y-Z transporter) is used to retrieve slides from a cassette 1 164 to load the processing
  • a slide gripper can move in the Z-direction lift the slides.
  • the slides are then moved in the X-direction and/or Y-direction.
  • the slides are then lowered into the processing apparatuses to place and remove slides from the processing apparatus.
  • the slide is routed to output cassette. If the processing apparatus does not perform coverslipping, slides can be routed to a coverslipper unit 1 170 (shown in phantom line).
  • the unit 1 170 is in the form of an imaging module adapted to obtain images for quality control within the system (such as to alert a user that a particular reagent or set of processing parameters is not providing an appropriate amount of staining) and/or patient diagnoses.
  • Information readers can be used to read information from the slides and/or coverslips. Readers can communicate this information to the controlling microprocessor(s), which microprocessor(s) will deliver reagents to particular liquid application stations in accordance with the specified protocol. The location and number of the code readers can be selected based on processing
  • the microprocessor(s) will also function to control the temperature of those portions.
  • Wastes from different stations can be directed to different containers, and some wastes (such as a solvent applied at multiple stations) can be directed to the same container while other wastes are directed to one or more containers, together or separately.
  • the waste removal systems can be fluidically coupled to a single container. Alternatively, each waste removal system can be fluidically coupled a separate waste container.
  • the stations or their components can include one or more thermal elements.
  • the thermal elements can include one or more heaters, coolers, or combinations thereof.
  • Thermal elements can include resistive heaters, radiant heaters, microwave heaters, ultrasonic heater or a Peltier device that can be used to raise or lower the temperature of a liquid (and a sample and a substrate as well).
  • two or more of these stations can include heaters (and/or coolers) that are independently controlled (to achieve the same or different temperatures/heating or cooling rates) or controlled together (to achieve substantially the same temperature/heating or cooling rate). It is also possible to have the heaters for several stations controlled together and heaters of one or more other stations controlled independently. Insulating segments or sections can be added between stations (such as between different staining stations) to provide thermal isolation.

Abstract

L'invention porte sur un appareil qui peut être utilisé pour appliquer et retirer des fluides pour traiter des échantillons biologiques. Un échantillon peut être mis en contact avec un fluide appliqué. Le fluide est distribué entre un premier substrat et un second substrat. Le premier substrat porte un échantillon. Une couche de fluide est formée dans un espace défini par le premier substrat et le second substrat. Le premier substrat se déplace par rapport au second substrat pour accumuler le fluide à proximité de la périphérie d'au moins l'un du premier substrat et du second substrat à l'aide d'une action capillaire. Le fluide accumulé est retiré de l'espace.
PCT/US2010/040413 2009-06-30 2010-06-29 Procédés et appareils d'application et de retrait de fluides pour traiter des échantillons biologiques WO2011002779A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US22204609P 2009-06-30 2009-06-30
US61/222,046 2009-06-30

Publications (2)

Publication Number Publication Date
WO2011002779A2 true WO2011002779A2 (fr) 2011-01-06
WO2011002779A3 WO2011002779A3 (fr) 2011-03-31

Family

ID=43411698

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2010/040413 WO2011002779A2 (fr) 2009-06-30 2010-06-29 Procédés et appareils d'application et de retrait de fluides pour traiter des échantillons biologiques

Country Status (1)

Country Link
WO (1) WO2011002779A2 (fr)

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102011050344A1 (de) * 2011-05-13 2012-11-15 Leica Biosystems Nussloch Gmbh Vorrichtung zur Handhabung von Objektträgern mit zwei Eindeckmodulen
DE102011050343B4 (de) * 2011-05-13 2013-06-20 Leica Biosystems Nussloch Gmbh Vorrichtung zur Handhabung von Objektträgern mit einem Lineartransport zum Transport der Racks
WO2013127990A1 (fr) * 2012-03-01 2013-09-06 Victorious Medical Systems Aps Procédé et système de répartition et d'agitation d'une quantité de liquide sur une lame de microscope
USD728120S1 (en) 2013-03-15 2015-04-28 Ventana Medical Systems, Inc. Arcuate member for moving liquids along a microscope slide
US9498791B2 (en) 2009-11-13 2016-11-22 Ventana Medical Systems, Inc. Opposables and automated specimen processing systems with opposables
CN110383034A (zh) * 2017-03-09 2019-10-25 霍罗杰克股份有限公司 自动化制备生物样本的系统及方法
US10634590B2 (en) 2014-03-11 2020-04-28 Emd Millipore Corporation IHC, tissue slide fluid exchange disposable and system
US10682245B2 (en) 2015-11-12 2020-06-16 The Provost, Fellows, Foundation Scholars, & The Other Members Of Board, Of The College Of The Holy & Undiv. Trinity Of Queen Elizabeth, Near Dublin Implantable biocompatible expander suitable for treatment of constrictions of body lumen
US10746752B2 (en) 2009-11-13 2020-08-18 Ventana Medical Systems, Inc. Opposables and automated specimen processing systems with opposables
US11484398B2 (en) 2019-11-22 2022-11-01 ProVerum Limited Implant delivery methods
US11602621B2 (en) 2019-11-22 2023-03-14 ProVerum Limited Device for controllably deploying expandable implants

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6544395B1 (en) * 1996-05-06 2003-04-08 Helena Laboratories Corporation System for the application of samples on a substrate
US20060120921A1 (en) * 2002-06-20 2006-06-08 Stuart Elliot Biological reaction apparatus with draining mechanism
US7186383B2 (en) * 2002-09-27 2007-03-06 Ast Management Inc. Miniaturized fluid delivery and analysis system

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6544395B1 (en) * 1996-05-06 2003-04-08 Helena Laboratories Corporation System for the application of samples on a substrate
US20060120921A1 (en) * 2002-06-20 2006-06-08 Stuart Elliot Biological reaction apparatus with draining mechanism
US7186383B2 (en) * 2002-09-27 2007-03-06 Ast Management Inc. Miniaturized fluid delivery and analysis system

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10746752B2 (en) 2009-11-13 2020-08-18 Ventana Medical Systems, Inc. Opposables and automated specimen processing systems with opposables
US9498791B2 (en) 2009-11-13 2016-11-22 Ventana Medical Systems, Inc. Opposables and automated specimen processing systems with opposables
DE102011050343B4 (de) * 2011-05-13 2013-06-20 Leica Biosystems Nussloch Gmbh Vorrichtung zur Handhabung von Objektträgern mit einem Lineartransport zum Transport der Racks
US9664701B2 (en) 2011-05-13 2017-05-30 Leica Biosystems Nussloch Gmbh System for handling slides having two coverslipper modules
DE102011050344B4 (de) 2011-05-13 2018-10-04 Leica Biosystems Nussloch Gmbh Vorrichtung zur Handhabung von Objektträgern mit zwei Eindeckmodulen
DE102011050344A1 (de) * 2011-05-13 2012-11-15 Leica Biosystems Nussloch Gmbh Vorrichtung zur Handhabung von Objektträgern mit zwei Eindeckmodulen
WO2013127990A1 (fr) * 2012-03-01 2013-09-06 Victorious Medical Systems Aps Procédé et système de répartition et d'agitation d'une quantité de liquide sur une lame de microscope
USD728120S1 (en) 2013-03-15 2015-04-28 Ventana Medical Systems, Inc. Arcuate member for moving liquids along a microscope slide
USD772424S1 (en) 2013-03-15 2016-11-22 Ventana Medical Systems, Inc. Arcuate member for moving liquids along a microscope slide
US10634590B2 (en) 2014-03-11 2020-04-28 Emd Millipore Corporation IHC, tissue slide fluid exchange disposable and system
US10881539B2 (en) 2015-11-12 2021-01-05 The Provost, Fellows, Foundation Scholars & The Other Members Of Board, Of The College Of The Holy & Undiv. Trinity Of Queen Elizabeth, Near Dublin Implantable biocompatible expander suitable for treatment of constrictions of body lumen
US10682245B2 (en) 2015-11-12 2020-06-16 The Provost, Fellows, Foundation Scholars, & The Other Members Of Board, Of The College Of The Holy & Undiv. Trinity Of Queen Elizabeth, Near Dublin Implantable biocompatible expander suitable for treatment of constrictions of body lumen
CN110383034A (zh) * 2017-03-09 2019-10-25 霍罗杰克股份有限公司 自动化制备生物样本的系统及方法
CN110383034B (zh) * 2017-03-09 2022-07-12 霍罗杰克股份有限公司 自动化制备生物样本的系统及方法
US11484398B2 (en) 2019-11-22 2022-11-01 ProVerum Limited Implant delivery methods
US11602621B2 (en) 2019-11-22 2023-03-14 ProVerum Limited Device for controllably deploying expandable implants

Also Published As

Publication number Publication date
WO2011002779A3 (fr) 2011-03-31

Similar Documents

Publication Publication Date Title
US9618430B2 (en) Thin film processing apparatuses for adjustable volume accommodation
WO2011002779A2 (fr) Procédés et appareils d'application et de retrait de fluides pour traiter des échantillons biologiques
EP2938994B1 (fr) Systèmes de traitement d'échantillon et procédés d'alignement de lames porte-objets
US9989448B2 (en) Specimen processing systems and methods for holding slides
AU2013369469B2 (en) Automated specimen processing systems and methods of using the same
US20110305842A1 (en) Floatable opposables for applying fluids to process biological samples
EP2939030B1 (fr) Systèmes de traitement d'échantillons et procédé de chauffage uniforme de lames
US10509216B2 (en) Specimen processing systems and methods for aligning slides
EP2938436B1 (fr) Systèmes et procédés de traitement d'échantillons permettant de modérer l'évaporation
US20220268673A1 (en) Specimen processing systems and methods for holding slides

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 10794643

Country of ref document: EP

Kind code of ref document: A2

NENP Non-entry into the national phase in:

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 10794643

Country of ref document: EP

Kind code of ref document: A2