cover element, method and treatment module to treat a biological sample on a substrate

专利摘要:
COVERAGE ELEMENT, METHOD AND TREATMENT MODULE TO TREAT A BIOLOGICAL SAMPLE ON A SUBSTRATE. Covering element for a substrate that supports a biological sample, which comprises the first and second opposite ends, the first and second opposite surfaces, a space on the second surface which, when juxtaposed with a substrate, forms a chamber and a fluid inlet towards the first end and in fluid communication with space. The space is limited by the walls of the space presenting one or more contour regions to improve the movement of the fluid within the chamber. A treatment module for a biological sample comprises the covering element, a support surface for a substrate carrying the biological sample and operable clamping device to removably retain the covering element in juxtaposition with the substrate for an incubation period. A method for incubating the biological sample with one or more reagents employs the cover element.
公开号:BR112014011935B1
申请号:R112014011935-0
申请日:2012-11-15
公开日:2020-11-03
发明作者:Mark Brian Dockrill；Anthony Favaloro；Kenneth Heng-Chong Ng；Martin Limon；Peter Toogood；Stephen John Bagnato
申请人:Leica Biosystems Melbourne Pty Ltd.；
IPC主号:

专利说明:

FIELD OF THE INVENTION
[001] This invention relates to instruments and methods for the automatic staining of anatomical pathology samples. It refers particularly, but not exclusively to a cover element, which forms a reagent chamber on a substrate, such as a slide on which a pathology sample has been placed. BACKGROUND OF THE INVENTION
[002] The instrumentation for the automated treatment of biological samples, such as anatomical pathology samples is well known. The treatment may include staining procedures of the types that are typical in immunochemistry, in situ hybridization, special staining and cytology. The automation of some staining procedures has increased the speed with which the test can be completed leading to early diagnosis and, in some cases, intervention. Staining is normally performed on samples placed on microscopic slides to enhance certain histological features in a biological sample, and incubation of the sample with small volumes of reagents is usually performed. In many cases, automated staining of samples involves manipulating the robotic arms to release an aliquot of the reagent to achieve staining. While automation has many advantages, there are also limitations associated with automating these procedures.
[003] In some cases, the coloring obtained by automated instrumentation is irregular or unreliable, which leads to the rejection or "failure" of some slides by the pathologist. The failure may be attributable to bubbles that form in the reagent that leads to irregular stains, and / or residues of reagents that produce inferior stains. In other cases, the cost to perform each test is prohibitively high, typically due to the high cost of purchasing and maintaining the instrumentation and / or the reagents used. In still other cases, stained areas are very small in relation to the sample size and are not useful for diagnostic analysis.
[004] The complexity of automated instrumentation can also be problematic with moving parts that require a myriad of calibration, maintenance and cleaning. In many cases, the yield of the processed sample is limited by batch processing regimes, where the sample processing time is limited by the slowest staining protocol to be administered to the batch.
[005] It would be desirable to improve the approaches available for the automated treatment of biological samples or at least provide a viable alternative to the methods and devices used.
[006] The discussion of the background of the invention included in this document with reference to documents, acts, materials, devices, articles and the like is intended to explain the scope of the present invention. It should not be taken as an admission or suggestion that any said material has been published, known or is part of common general knowledge in the field of patents as on the priority date of any of the claims. SUMMARY OF THE INVENTION
[007] Seen from one aspect, the present invention provides a covering element for a substrate that supports a biological sample, the covering element comprising: a) first and second opposite ends; b) first and second opposite surfaces; c) an empty space on the second surface that, when juxtaposed with a substrate, forms a chamber; and d) a fluid inlet to the first end and in fluid communication with the void; where the empty space is bounded by empty walls having one or more contour regions to improve the movement of the fluid inside the chamber.
[008] Preferably, the cover element includes a fluid outlet to the second end and in fluid communication with the vacuum and through which the fluid can be withdrawn.
[009] In one or more modalities, one or more contour regions comprise rounded corners that connect the side walls of the empty space with an end wall. In one embodiment, the contour regions can comprise the rounded corners for the second end of the cover member to encourage removal of fluid from the chamber. In another embodiment, the contour regions may comprise the rounded corners that connect the side walls of the void with an end wall towards the first end of the cover element to stimulate the flow of fluid within the chamber. In yet another embodiment, one or more contour regions may comprise rounded frames that connect the walls of the void to an empty ceiling on the second surface of the covering element. In another embodiment, one or more contour regions may comprise a cone or the final region similar to a flat cylinder that joins the opposite side walls of the void.
[0010] The cover member can provide a volume when the chamber is closed, for example, from 30 to 200 pL, preferably from 50 to 150 pL and more preferably from about 100 pL to 125 pL. In one or more embodiments, the chamber has a height of 50 to 200 pm. In some embodiments, the height is preferably 100 to 150 pm. In certain embodiments, the cover element includes an inlet reservoir, which has a volume sufficient to receive one or more aliquots of a fluid to be distributed in the chamber during a stage of a treatment protocol.
[0011] The second surface has a vacuum ceiling that, in several modalities, has a coating that increases the spread of the reagent from the inlet to the outlet. The finish can be, for example, the texture selected from a group that includes: embossed, wavy, dimpled, slanted, curved and wavy. Alternatively, the cladding may be a finishing material or cladding for at least part of the ceiling and / or walls of the void.
[0012] Preferably, the covering element is adapted to be kept in juxtaposition with the substrate during a treatment protocol. In some embodiments, the cover element is either disposable or semi-disposable (for example, used for 5, 10, 15 or 20 protocols before being replaced). In other embodiments, the cover element is formed from at least two parts, including a cover element body and a cover element insert, where the cover element insert is configured to form the chamber with the substrate. In this arrangement, the cover insert can be disposable.
[0013] In some embodiments, the covering element includes a moisture barrier configured to reduce the drying of a sample on a substrate with which the covering element is used. The moisture barrier can take any suitable shape that does not interfere with the sample on the substrate. For example, the moisture barrier may be a mantle material adapted to cover, but not come in contact with the sample on the slide. Alternatively, the moisture barrier can be a vapor barrier that prevents the sample on the substrate from dehydrating.
[0014] In one embodiment, the cover element includes a guiding device, at the entrance, configured to direct the fluid into the entrance. Preferably, the guiding device comprises a neck shaped to receive a correspondingly shaped distribution probe tip, so that they form combined contact for forced delivery of a fluid from the probe into the inlet. Thus, the neck can have a decreasing taper towards the second surface that can accommodate the probe tip. Ideally, the guide device is configured to form a tight fit with the end of a distribution probe. This can be achieved by providing the guide device with sufficient suitability to receive and form a seal with the end of a dispensing probe, although in another arrangement of the tip the probe is compatible.
[0015] In another embodiment, the cover component has a distribution edge arranged in fluid communication with the inlet. In use, the cover member is adapted to pivot around the distribution edge and the pivoting movement causes fluid movement in the inlet from the distribution edge towards the outlet. The cover part can be further adapted to articulate about an axis extending through it and perpendicular to a plane that extends orthogonally between the first and second ends, where the rotation about said axis tilts the cover element . It may be desirable to tilt the cover element to prevent premature release of fluid at the inlet, or to have access to a blade under the cover element, when in an open condition.
[0016] In yet another embodiment, the cover element includes a fluid delivery feature configured to disperse fluid from the inlet to at least one width of the chamber formed in the cover element. Preferably, the fluid delivery feature comprises a channel that spans the width of the chamber. In one embodiment, the channel has a stepped profile with increasing height towards the first end of the covering element. Ideally, the channel is configured to store a volume of fluid from the inlet. The stored fluid feeds a fluid front which is gradually spread over the substrate.
[0017] The fluid distribution functionality can be configured to disperse the fluid in a closed state or in an open state. For open dispensing, the fluid dispensing functionality is configured to disperse the fluid during the relative sliding movement of the cover element and the substrate from an opening condition, in which the sample is outside the chamber, to a condition closed in which the covering element covers at least a portion of the sample on the substrate, thus dragging the fluid from the dispensing functionality along the surface of the substrate. In a closed state, the covering element overlaps at least a portion of the sample on the capillary-acting substrate and draws the fluid from the distribution functionality along the substrate surface.
[0018] The covering element may further comprise a sliding guide device configured to guide the substrate during the relative sliding movement of the covering element and the substrate between the open and closed conditions. Ideally, the sliding coating element also includes a moisture barrier configured to reduce drying of a sample on a substrate with which the covering element is used. The moisture barrier can be a physical material barrier or a vapor or other barrier adapted to minimize drying of the sample.
[0019] Seen from another aspect, the present invention provides a treatment module for a biological sample, the module comprising: e) a cover element according to any one of the preceding claims; f) a support surface for a substrate that has a respective biological sample; and g) operable fixing devices to retain the removable cover element in juxtaposition with the substrate for an incubation period.
[0020] The clamping device applies sufficient clamping force to prevent the reagent from escaping from the space between the substrate and the covering element during a protocol, while not damaging or breaking the substrate. Clamping forces can be in the range, for example, from about 3 N to 300 N. In some cases, higher clamping forces may be difficult to obtain, for example, when a plurality of treatment modules are incorporated into an automated instrument. Thus, it may be desirable to use a lower clamping force, for example, from 250 N to 100 N. Clamping forces as low as 10 N can also be used. In one form, the clamp comprises an inclined resilient component to retain the cover element in juxtaposition with the substrate. In various embodiments, the treatment module also offers substrate retention means configured to retain the substrate on the support surface during opening of the chamber, for example, to overcome the adhesion forces.
[0021] In one or more modalities of the support surface it comprises a heat exchanger configured to control the temperature of a biological sample of the substrate during a treatment protocol. It should be understood, however, that the heat exchanger can be part of a cover element described above, or can be coupled to a cover element.
[0022] Typically, the treatment module includes a robot configured to position one or both of the substrate and the cover element of the treatment module, and can also be configured to deliver reagent to a cover element entry during a treatment protocol. . In various embodiments, the treatment module includes an operable coupling to alternately connect one or more outlets of the cover element with an opening to the atmosphere and one or more respective sources of negative pressure. Typically, one or more sources of negative pressure generate a controlled vacuum between -2 kPa and -15 kPa. One or more negative pressure sources can be controlled by a controller device programmed to apply negative pressure over a period of, for example, 1,000 ms to 5,000 ms, and preferably for about 2,000 ms to 3,000 ms.
[0023] The treatment module can be configured for use with an automatic sample processing instrument that comprises a plurality of operable treatment modules, independently under the control of an instrument controller. Ideally, the fixing devices, heat exchanger, robot, negative pressure sources and liquid distributors and other components with which the treatment module is used are also under the control of the instrument controller.
[0024] In one embodiment, the treatment module includes an articulation device configured to rotate the cover element over a distribution end over the cover element, causing fluid at the inlet to move from the distribution end towards the outlet , and where the hinge means can be operated to rotate the cover element from an open condition and a closed condition in which the cover element and the substrate are juxtaposed to form a chamber.
[0025] Preferably, the articulation device is an operable articulation arm for positioning the cover element in the open position, where a distribution edge of the first end of the cover element is in contact with the substrate and the second surface is arranged with an angle of 1 to 20 ° in relation to the substrate. The articulation devices can also be operable to stir the reagent inside the chamber. In one embodiment, the articulated arm is operated to position the cover element in the open position so that the substrate and the second surface are angled to receive an aliquot of the fluid at the entrance of the cover element. In one embodiment, the substrate and the second surface are arranged at an angle of about 5 to 60 degrees. In one embodiment, the substrate and the second surface are arranged at an angle of about 8 to 25 °. In one embodiment, the substrate and the second surface are arranged at an angle of approximately 10 °. The rotating arm can also be operable to arrange the module in a state of release in which the covering element and the substrate are dissociated, and / or to cause the covering element to tilt on an inclination axis that extends through the cover element and perpendicular to a plane extending orthogonally between the first and second ends. The slope can provide access to the substrate within the treatment module, and / or can prevent premature release of fluid from the inlet to the chamber. In one form, tilt means are provided for polarizing the tilt direction of the cover element over the tilt access.
[0026] The treatment module can also include a washing compartment to expose the second surface of the cover element to a washing reagent. Thus, the support surface can be shaped to receive a substrate having a sample on it and, in the absence of a substrate, to form the wash compartment.
[0027] In one embodiment, the treatment module includes a trigger for slidingly moving the covering element and the substrate between an open condition in which the sample is not covered by the covering element, and a closed condition, in which, at least part of the sample is covered in a chamber formed by a covering element and the substrate. The treatment module can also include a moisture barrier, as described above.
[0028] Seen from another aspect, the present invention provides a covering element for a substrate that supports a biological sample, the covering element comprising: a. first and second opposite ends; B. first and second opposite surfaces; ç. a vacuum on the second surface which, when juxtaposed with a substrate, forms a chamber; d. a fluid inlet towards the first end and in fluid communication with the void; and. a fluid outlet towards the second end and, in fluid communication with the empty space; and f. guide device at the entrance, configured to direct the fluid into the entrance.
[0029] The guiding device may comprise a neck shaped to receive a distribution probe tip correspondingly. The neck may have a decreasing taper in the direction of the second surface and / or fit. In any case, it is desirable that the guide device be configured to form a tight fit with the tip of the distribution probe.
[0030] Seen from yet another aspect, the present invention provides a method for incubating a biological sample with one or more reagents employing a cover element with a guiding device, which includes the steps of: a. provision of the sample on a substrate; B. positioning the substrate and the covering element to form the chamber; ç. positioning a tip of the distribution probe in combination contact with the fluid inlet; and d. directing a first volume of a first reagent into the inlet, with sufficient force for the first reagent to substantially cover the sample on the substrate.
[0031] The first reagent can be forced into the entrance of a positive pressure pump, such as a syringe plunger or a gear pump, coupled to the tip of the distribution probe.
[0032] The method may alternatively and additionally include the steps of: a. provision of the sample on a substrate; B. positioning the substrate and the covering element to form the chamber; ç. positioning a tip of the dispensing probe to dispense reagent inside the fluid inlet; and d. dispensing at least the second volume of a second reagent into the inlet.
[0033] The method can also include the application of a negative pressure on the outlet to drag the reagent inside the chamber towards the outlet. Typically, the first reagent (being a reagent that is delivered through the inlet, with a driving force), is a high value reagent, while the second reagent (being a reagent that is dispensed into the inlet without a driving force) is a low value reagent. The method may further include the step of tilting the cover member to raise the outlet limiting or thereby preventing premature release of the reagent from the inlet into the chamber.
[0034] Seen from yet another aspect, the present invention provides a covering element for a substrate that supports a biological sample, the covering element comprising: a. first and second opposite ends; B. first and second opposite surfaces; ç. an empty space on the second surface which, when juxtaposed with a substrate, forms a chamber; d. a fluid inlet to the first end and, in fluid communication with the void, and; and. a fluid outlet to the second end and, in fluid communication with the void; f. an edge arranged in fluid communication with the distribution inlet; where the cover member is adapted to pivot around the distribution edge, and where, in use, said pivoting movement causes fluid to flow in from the distribution edge towards the outlet.
[0035] The covering element can also provide a moisture barrier configured to reduce the drying of a sample on a substrate, with which the covering element is used, as described above. Likewise, the cover member can provide a reservoir at the entrance with a volume sufficient to receive one or more aliquots of a reagent.
[0036] Seen from another aspect, the present invention provides a treatment module for a biological sample, the module comprising: a. a cover element having a support edge; B. a support surface for a substrate that has a respective biological sample on it; and c. articulation device configured to articulate the covering element over the distribution edge causing fluid to pass from the inlet along the substrate from the distribution edge towards the outlet; where the hinge device is operable to articulate the cover element for an open condition and a closed condition in which the cover element and the substrate are juxtaposed to form a chamber.
[0037] The means of articulation can take any suitable shape. In a preferred embodiment, the articulation means comprise an operable articulating arm for positioning the cover element in the open position, where a distribution edge of the first end of the cover element is in contact with the substrate and the second surface is arranged with a 1 to 20 degrees angle to the substrate. Preferably, the articulated arm is operated to position the cover element in the open position, so that the substrate and the second surface are arranged at an angle of approximately 10 degrees, to receive an aliquot of the reagent at the entrance of the cover element. The pivotable arm can also be operated to cause the cover element to tilt over an inclination axis that extends through the cover element and perpendicular to a plane that extends orthogonally between the first and second ends. The articulation device can also be operable to position the module in a release state in which the covering element and the substrate are dissociated and / or to stir the reagent inside the chamber.
[0038] The treatment module may also include a tilt device to tilt the tilt direction of the cover element around the tilt access and / or a substrate holding device configured to removably retain the substrate on the support surface during separation of the covering element and the substrate. The substrate retaining device may comprise a resilient element configured to removably retain the substrate on the support surface with a force sufficient to overcome an adhesion force between the covering element and the substrate during separation.
[0039] Ideally, the treatment module also includes fixing devices for removably retaining the cover element in the closed position. A washing compartment can also be provided to expose the second surface of the cover element to a washing reagent. In one embodiment, the support surface is shaped to receive a substrate having a sample in it and, in the absence of a substrate, forms the washing compartment. The treatment module can also provide a moisture barrier. In one or more embodiments, the treatment module has an operable coupling to alternately couple one or more outlets of the cover element with one or more respective sources of negative pressure.
[0040] Seen from yet another aspect, the present invention provides a method for incubating a biological sample with one or more reagents using a treatment module as described above, including the steps of: a. provision of the sample on a substrate; B. positioning the substrate and the covering element in an open condition, in which the covering element is tilted in such a way that the distribution edge contacts the substrate; ç. dispensing a first reagent into the inlet; and d. hinge of the cover element for the closed condition, the hinge action causing the dispensed reagent to substantially cover the sample on the substrate.
[0041] Ideally, the articulation action is controlled at a rate that increases the capillary flow of the reagent to substantially cover the sample on the substrate. A negative pressure applied to the outlet can assist in the preparation of reagents inside the chamber towards the outlet. A negative pressure can be used to evacuate and / or stir the fluid in the chamber. Several steps can be achieved using a controller according to a pre-programmed hinge action, which increases the flow of reagent over the substrate for a plurality of reagents and / or for a plurality of protocols for the treatment of a sample.
[0042] The method may also include the step of removing the blade from the support surface and immersing the second surface of the cover element in a washing reagent.
[0043] Seen from another aspect, the present invention provides a covering element for a substrate that supports a biological sample, the covering element comprising: a. first and second opposite ends; B. first and second opposite surfaces; ç. a space on the second surface for forming a chamber with the substrate; d. a fluid inlet towards the first end and, in fluid communication with the space; and is. a fluid delivery feature from which the fluid is delivered; where the fluid delivery functionality is configured to deliver fluid from the inlet to at least one width of the substrate.
[0044] In one embodiment, the fluid distribution functionality comprises a channel that covers the width of the chamber. The channel can have a stepped profile with increasing height towards the first end of the cover element and can be configured to store a volume of fluid from the inlet, where the volume of the stored fluid feeds a fluid front, which is gradually spread over the substrate. The cover element can also provide an outlet for the second end of the cover element, through which the fluid can be drawn.
[0045] Preferably, the fluid distribution functionality is configured to supply the fluid during the relative sliding movement of the cover element and the substrate from an open condition, where the sample is outside the chamber, for a condition closed in which the cover element covers at least a portion of the sample on the substrate, thereby dragging the fluid from the fluid delivery functionality along the substrate. This can be called "open dispersion".
[0046] Alternatively and in addition the fluid distribution functionality is configured to supply fluid in a closed condition, in which the covering element overlaps at least a portion of the sample on the substrate, where said dispersant uses capillary action to drag fluid from the fluid distribution functionality along the surface substrate. This can be referred to as "closed distribution".
[0047] The covering element may further comprise a sliding guide device configured to guide the substrate during the relative sliding movement of the covering element and substrate between the open and closed conditions. A moisture barrier configured to reduce drying of a sample on a substrate with which the cover element is used can also be provided.
[0048] Seen from another aspect, the present invention provides a method for the incubation of a biological sample with one or more reagents with a cover element that has a fluid distribution functionality comprising the steps of: a. provision of the sample on a substrate; B. positioning the substrate and the covering element in an open configuration, in which at least a portion of the end of the substrate is arranged in juxtaposition with the second surface of the covering element in the region of the fluid distribution functionality; ç. distribution of a reagent at the entrance and using capillary action to drag the reagent along the substrate.
[0049] Preferably, the method includes sliding the substrate and the covering element with respect to the other substrate and the covering element, from an open condition, where the sample is outside the chamber, to a closed condition in the which the covering element covers at least a portion of the sample inside the chamber, where said sliding action drags the reagent from the dispensing functionality along the substrate. Ideally, the sliding action is controlled at a speed that increases the flow of reagent to substantially cover the sample on the substrate. The fluid can be distributed into the inlet, while the substrate and the covering element are in the open condition ("open distribution") or after they are in a respectively closed condition ("closed distribution").
[0050] A vacuum can be applied to extract reagent through the chamber from the inlet to the outlet in order to assist in the distribution of the fluid, or to evacuate, or stir the fluid in the chamber. In one embodiment, the method includes masking the substrate to limit drying of the sample, when the covering element and the substrate are reopened.
[0051] Seen from yet another aspect, the present invention provides a module for treating a biological sample, the module comprising: a. a cover element having a fluid distribution functionality; B. a support surface for a substrate that has a respective biological sample; ç. a linear movement device for the sliding movement of the covering element and the substrate between an open condition in which the sample is not covered by the covering element, and a closed condition, in which at least part of the sample is covered with a chamber formed by the covering element and the substrate.
[0052] In one or more embodiments, the treatment module includes a washing compartment to expose the second surface of the covering element of a washing reagent during a washing step of a treatment protocol with the treatment module. A moisture barrier can also be provided to protect the sample. The treatment module may also provide an operable coupling for alternately coupling one or more outputs of the cover element with one or more respective sources of negative vacuum pressure generation. BRIEF DESCRIPTION OF THE DRAWINGS
[0053] Modalities of the invention will now be described in more detail, by way of example only, with reference to the accompanying drawings. It should be understood that the modalities shown are only examples and may not be scaled, in all cases. The examples discussed are not to be considered as limiting the scope of the invention as defined in the appended claims. It should be understood that the parts described are numbered in series (for example, 1,000, 2,000, 3,000), where the same numbers designate equal parts in general.
[0054] Figure 1 is a schematic isometric view of a cover element according to an embodiment of the invention.
[0055] Figure 2 is a side view of the cover element of Figure 1 which also shows a substrate in the form of a blade.
[0056] Figure 3 is a schematic illustration of the second surface (underside) of the covering component of figures 1 and 2.
[0057] Figures 4a to 4e are schematic illustrations that represent variations in the form of entry of a cover element according to an embodiment of the invention. Figures 4a, c and d represent an end sectional view through the entrance. Figure 4b represents a top view of the cover member of Figure 4a and Figure 4e represents a top view of the cover member of Figures 4c and d.
[0058] Figures 5a through 5c are schematic illustrations of the cover element body, a cover element insert and a cover element (composed of a body cover element, in combination with a cover element insert), respectively .
[0059] Figures 6a to 6c are schematic illustrations of a treatment module according to an embodiment of the invention, with the cover element in the closed position (figure 6a) and an open position (figures 6b and 6c).
[0060] Figures 7a and 7b are schematic sectional views of a treatment module according to another embodiment of the invention, with a removable cover element insert and washing compartment.
[0061] Figure 8 is an isometric view of a cover element according to another aspect of the present invention.
[0062] Figures 9a to 9c are seen in end, side and bottom section of the cover element of figure 8.
[0063] Figures 10a to 10d show a simplified sectional view of a cover element according to another embodiment of the invention, in open, dispensing, closed and released conditions respectively, with respect to the blade.
[0064] Figure 11 is another schematic illustration of a cover element according to an embodiment of the invention.
[0065] Figure 12 is a schematic illustration of the elements of a treatment module for use with a cover element, of the type illustrated in figures 8 to 11.
[0066] Figure 13 is a side sectional view of a treatment module, of the type illustrated in figure 12.
[0067] Figure 14 is an isometric view of elements of a treatment module according to an embodiment of the invention.
[0068] Figure 15 is a side view of the treatment module in figure 14.
[0069] Figure 16 is a schematic bottom view of a cover element according to another aspect of the present invention.
[0070] Figure 17 is an isometric view of the cover element of Figure 16 with a substrate in the form of a pathology slide, in an open state.
[0071] Figure 18 is a sectional view of the cover and substrate element of figure 17.
[0072] Figure 19 is an enlarged sectional view showing the fluid and blade distribution functionality in Figures 16 to 18.
[0073] Figure 20 is an isometric view of the covering element and substrate of figures 16 to 19 in a closed condition.
[0074] Figure 21 is a side sectional view of the covering element and the substrate of figure 20.
[0075] Figure 22 is an isometric view of the covering element and substrate of Figure 17, with a moisture barrier, in the form of a physical envelope.
[0076] Figure 23 is an isometric view of the components of a treatment module for use with the cover element of figures 16 to 22.
[0077] Figure 24 is an example of an instrument for automatic sample processing with which modalities of the invention can be used.
[0078] Figure 25 is a schematic illustration of a controller for the instrument in Figure 24. DETAILED DESCRIPTION
[0079] It is desirable to incubate small volumes of reagents on a substrate such as a microscope slide. Samples can be treated, while the slides are held in a slide tray or individually in sample treatment modules.
[0080] Referring first to figure 1, a covering component 1,000 according to an embodiment of the invention is shown, for use with a substrate 200 (shown in figure 2) to support a biological sample. To facilitate consultation, substrate 200 will hereinafter be referred to as "blade" 200. The cover member has a first end 1010 and a second end 1020 and a first surface 1110 and a second surface 1120. A vacuum 1124 is formed on the second surface, defined by a vacuum limit in the form of walls 1122 and an empty space 1140.
[0081] Figure 2 is a side sectional view of the cover element 1000 and blade 200 arranged in juxtaposition, to form a chamber 1300. Fluid inlet 1012 is provided at the first end of the cover element and a fluid outlet 1022 is provided at the second end of the cover element. The entrance and exit are in fluid communication with the empty space 1124, in order to allow a reagent to enter the chamber through the entrance and exit through the exit 1022. A guide device 1014 is also provided at the entrance.
[0082] In a preferred embodiment, the covering element 1000 is configured to be used in an automated sample processing instrument 7000, such as the type illustrated in figure 25. US Provisional Patent Applications number 61 / 560,569, entitled " Biological Sample Treatment Apparatus "and 61 / 560,559, entitled" An Automated System and Method of Treating Tissue Samples on Slides "both filed simultaneously with this application on 16 November 2011 by the same applicant describe such instruments and the content of those applications is incorporated into this document as a reference.
[0083] The instrument uses a robotic arm to dispense a reagent at the entrance of the cover element. The guiding device 1014 directs a delivery probe 400 from the apparatus to the entrance, such that the robotic controller does not need to accurately locate the probe tip 410 into the entrance well. Instead, the controller only needs to position the probe tip 410 inside the inlet opening 1013 and the guide device 1014 directs reagent dispensed from the probe tip through the inlet 1012 and into the chamber 1300.
[0084] In a preferred embodiment, the guiding device is configured for reagent distribution contact into the inlet. Thus, the guide device 1014 comprises a neck 1016, which is shaped to receive a correspondingly shaped distribution probe tip 410 (Figure 2). The neck can be reduced to form, for example, an angle of 45 degrees with respect to an axis that passes through the entrance and receives a tip of the probe 410 of correspondingly shaped distribution having an angle of 45 ° between the axis that extends through the probe 400 and the outer probe tip walls. The probe tip 410 correspondingly shaped and the neck 1016 cooperated to form a coupling interface between the probe tip and the neck for dispensing a reagent.
[0085] In one or more modalities, the neck presents suitability, so that the coupling interface provides a perfect fit between the tip of the probe and the inlet neck to substantially prevent the leakage of forced reagent into the inlet through positive pressure . However, the use of a gasket or seal ring on the coupling interface is also contemplated. Suitability can be provided by a material property of the covering element, including the neck, for example, when the covering element is manufactured from a compatible material. Alternatively, there may be a coating of material compatible with the neck area of the cover element or the tip of the probe.
[0086] During the dispensing of the high value reagent, it is desirable that the tip of the dispensing probe 410 be placed in combination contact with the neck 1016, as described above. However, such contact may not be necessary for the release of less expensive volume fluid reagents, such as DI water, alcohol, a degreasing solution and the like. This is particularly the case when there is excess distribution (that is, distribution of more than one aliquot of reagent) or cleaning. In one embodiment, cleaning involves dispensing a cleaning reagent without contact into the inlet and then withdrawing the reagent, for example, using a vacuum, back through the inlet or through the outlet when one is provided.
[0087] The distribution probe can be, for example, a Fluid Transfer Probe (FTP) 7028 robot (using a temporary or permanent pipette tip) or a Volume Fluid Robot (BFR) 7 014 from an automated instrument 7000, such as the type shown in figure 24. In one embodiment, FTP or BFR can also be used to position a cover element 1000 on a blade 200 so that they form chamber 1300 between them. In figure 24, a plurality of cover elements are shown on the individual sample treatment modules 7012 inside the 7000 instrument. Each of these can be controlled independently, so that the instrument transfer rate for individual treatment modules 7012 it is not limited by the incubation time required for protocols to be performed on the other modules in the device.
[0088] In this provision, the instrument may have reduced complexity since a dedicated robot for the placement of the cover element is not necessary. Since the covering element is arranged in juxtaposition with a slide having a biological sample placed on it, it is fixed in position using any suitable means and does not move for the duration of the treatment protocol. In figure 2, arrow C designates the direction of a clamping force applied to the cover element to maintain its position throughout the protocol.
[0089] Advantageously, once the covering component 1000 is positioned and fixed in place, the covering element does not need to move in relation to the cursor 200 for the duration of the protocol. The use of positive pressure to force the reagent into the chamber and / or a vacuum to drag the reagent through the chamber is sufficient to complete most protocols. Since the treatment protocol can be completed without moving the relative position of the covering component 1000 and the blade 200, there is minimal exposure of the sample to atmospheric air. Therefore, the risk of dehydration of the sample is low and at the conclusion of a given protocol, the sample can be covered by the slides for transport and / or further processing.
[0090] Reagents can remain inside the chamber for an incubation period, before being removed via outlet 1022. During incubation, the temperature of the sample (and the reagent) can be modified, for example, by heating or cooling of a heat exchanger related to the treatment module. The heat exchanger is normally supplied in the form of a 5300 heating / cooling block (figures 6a to 6c). Ideally, the heat exchanger has the ability to vary the temperature of the sample (and the reagent in the chamber) in the range of 20 to 95 ° C, although higher temperatures (above 120 ° C, for example) may be necessary for some protocols. Some reagents can lead to the formation of bubbles during the heating steps. Typically, bubbles migrate towards inlet port 1012 and / or outlet port 1022, when vented into the atmosphere. The rate of temperature change can be critical to the effectiveness of the protocol, for example, in PCR where fast transitions are needed. Ideally, the heat exchanger accommodates these changes and also has the ability, in one or more modes, to cool. In various aspects of the present invention, the heat exchanger is illustrated as a heating / cooling pad positioned below the blade. It should be understood, however, that the heat exchanger can be coupled or incorporated into the cover component, in various modalities. For example, the cover element can comprise a metal block with high thermal mass, such that it can heat and actively cool the samples (for example, by refrigeration). Alternatively, the heating means may comprise heating pads, RF, microwave, and / or convection means and the cooling means may comprise a cooling medium, fins and / or a Peltier effect refrigerator. In other embodiments, the covering element can heat and / or cool and the substrate support heats and / or cools in combination.
[0091] Typically, the high-value reagent is forced into the inlet in "contact mode" (ie, with the probe tip in contact with the inlet), using a positive pressure pump, such as a syringe plunger. Preferably, the operation of the syringe pump is under the control of a controller 7060 associated with the automated instrument 7000. Thus, once the tip of probe 410 is received in an adjusted manner inside the neck 1014, the syringe pump is activated to dispense an aliquot of reagent into the chamber. With this approach, actively moving the reagent into the chamber using positive pressure this minimizes the amount of reagent needed, and the time for the reagent to enter the chamber and cover the sample on the slide.
[0092] During forced release of the reagent into chamber 1300, outlet 1022 is vented to atmospheric pressure. Controlling the forced release rate provides fluid flow management as it moves over the blade, thereby minimizing the risk of bubble formation within the chamber. In some protocols, the reagent can be particularly viscous and the spread of the reagent over the slide surface inside the chamber can be assisted by applying a vacuum to outlet 1022. After the incubation period, the reagent can be evacuated from the chamber by applying a vacuum at the outlet or by washing with the injection of an excess of reagent. The arrow F (figure 2) designates the flow direction of the reagents distributed into the chamber. In order to provide the required pressure gradient across the chamber, a valve (not shown) can be provided and is operable to switably connect the outlet to atmospheric air or a negative pressure source.
[0093] A typical treatment protocol involves the distribution of fluid reagents by volume inside the chamber to wash or otherwise treat the sample. During a washing step, it is desirable to clean the inlet 1012 to remove any high-value residual reagent that has adhered to the inlet walls, for example, during forced delivery of a high-value reagent, in contact mode. Thus, a probe dispensing fluid reagents in volume at port 1012 does not need to make combination contact with the 1014 guide / neck device. In several stages of a protocol it may be desirable for certain reagents to be dispensed in "non-contact mode" in such a way the coupling surface to be washed.
[0094] Figures 4a to 4e illustrate examples of different air inlet profiles for the covering element 1000. Figures 4a, 4c and 4d represent sectional end views through inlet 1012. Figure 4b represents a top view of the cover element of figure 4a, and figure 4e represents a top view of the cover components of figures 4b and 4c. As shown in figures 4b and 4e, outlet 1022 may display the top of the cover element (i.e., through the first surface) or through the second end of the cover element as in figures 1 to 3, or, for example, by means of a front or rear surface of the cover element.
[0095] Figure 4a shows a variation of the entrance profile of the cover element of figures 1-3, where the guide device 1014 is extended to accommodate a greater volume of reagent, thus forming a reservoir 1018. Similar reservoirs 1018 are shown in air inlet profiles of figures 4c and 4d. Reservoir 1018 has a volume sufficient to store more than one aliquot of reagent. An advantage of providing a 1012 inlet to the reservoir is to allow mixing of the various reagents before entering the 1300 chamber. Another advantage is that storing multiple reagent dispensers can reduce the load on the dispensing robots used in an automated instrument thereby reducing the waiting time between steps in a protocol. In addition, the larger elliptical openings in figures 4c and 4d, reduce the complexity of the movements performed by automated instrumentation robots to position the nozzles of the reagent dispenser since the target area of the distribution is larger. To attenuate the premature release of reagent from the reservoir into the 1018 chamber, a treatment module with which the cover element is used can be adapted to tilt the cover element to raise the outlet thus avoiding the release of reagent into the chamber.
[0096] Figure 3 is a schematic illustration of the second surface (underside) of the covering component of figures 1 and 2. Figure 3 shows boundary walls contoured in 1126. Contoured boundaries 1126 in relation to the first auxiliary end with fluid flow inside the chamber 1300. bypassed limits 11261 'to the second end 1020 militate against reagent and / or reagent debris remaining inside the chamber after washing or evacuation. Evacuation can be achieved, for example, by activating a source of negative pressure (i.e., vacuum), coupled to outlet 1022 to remove or clean reagent from the chamber. Although in some modalities the boundary limits may have the same geometric shape, it should be noted that this need is not the case. For example, in figure 3, the boundary boundaries 1126 have a smaller radius than the boundary boundaries 1126 '. In another embodiment (not shown) the contour boundaries can be merged to form a tapering of one end (or both ends) of the void, so that the void does not include a head shape at either (or both) ends with a flat arrow or similar to a flat cylinder. However, the inclusion of such a taper can reduce the area of the slide covered by the chamber and thus limit the effectiveness of the reagents distributed into the slide staining chamber.
[0097] In figure 3, an inlet 1012 has a similar diameter to outlet 1022, although this need is not the case. As can be seen in figure 4b, the diameter of the entrance opening to the void can be greater than the diameter of the exit exiting the void.
[0098] Figures 5a to 7b show an alternative embodiment of a covering component 1000, which consists of two parts: body of the covering element 1100 (figure 5a) and insertion of the covering element 1200 (figure 5b). Figure 5c shows the cover element body and the insertion of the cover element together. Meanwhile, the body of the cover element 1100 has grooves 1150 where the opposition of the tongue portions 1250 of the insert of the cover element 1200 is received slidingly.
[0099] Inlet 1012 in the cover element body 1100 is arranged to couple with inlet extension 1012 'in the insertion of the cover element. Likewise, the outlet 1022 on the cover element body is configured to couple with the outlet extension 1022 'in figure 5b. Coupling the inlet / inlet extension and outlet / outlet extension in this way facilitates the distribution of a reagent where the chamber formed by the insertion of the cover element 1200 having an empty space 1124 which, when juxtaposed with a blade 200 (figure 6a to 6c) forms a reagent chamber. While the arrangement shown in figures 5a to 5c provides a coupling between the body of the cover element and the insertion of the cover element that allows sliding engagement, it should be understood that other arrangements are also contemplated, such as, for example, magnetic couplings and suction between the elements comprising a cover element.
[00100] Figures 6a to 6c show a treatment module 5000 according to an embodiment of the invention. Treatment module 5000 includes a support surface 5100 on which a blade 200 is supported. Optionally, a heat exchanger in the form of a 5300 heating / cooling block (as described above) is provided between the support surface 5100 and the blade 200 to change the temperature of the reagents inside the chamber during a treatment protocol. The blade 200 fits below the second surface 1120 of the covering element / insert of the covering element. Figures 6a to 6c further show a drive arm 5110 for positioning the covering element 1000 in juxtaposition with the blade 200. A clamping element 3200 is provided to retain the covering element 1000 and blade 200 in juxtaposition for the duration of a protocol. of treatment. The fastening element 3200 can be, for example, a torsion spring, which exerts a force on the covering element 1000.
[00101] Although the illustrated embodiment shows the drive arm 5110 positioned on the longest side of the cover element 1000, it should be understood that the drive arm can also be located at one end of the cover element. Thus, the arm 5110 can be operable to open and close the cover element 1000 longitudinally.
[00102] In addition to performing advanced staining protocols, a cover element 1000 that incorporates a removable / replaceable cover element insert 1200 can be useful in applications involving Polymerized Chain Reaction (PCR) protocols. In these protocols, transition of debris from one protocol to another can lead to contamination and insufficiency of samples. Therefore, it is necessary to clean completely or otherwise prevent the passage from one test to the next. Thus, incorporating a disposable removable cover element insert 1200 into the covering component 1000 can eliminate or at least reduce the risk of transit debris or cross-contamination and therefore may be desirable for applications such as PCR.
[00103] Figures 7a and 7b are schematic representations of another embodiment of the cover element also showing the driving arm 5110. Cover element body 1100 is shown with a cover element insert 1200. Figure 7a shows treatment module 5000 with a blade 200 held on a heating pad 5300 on the support surface 5100. In figure 7b, the blade 200 has been removed and the surface 5310 and walls 5320 of the heating pad 5300 form a washing compartment 5500. Thus, since the blade 200 has been removed after the completion of a treatment protocol, the second surface of the cover element (or insertion of the cover element) can be immersed into the washing compartment 5500 for cleaning. The cleaning reagent can be dispensed through the inlet of the cover component 1012/1012 'and removed through the outlet 1022/1022'. Alternatively, the cleaning reagent can be dispensed directly into the 5500 wash compartment and drained through one of the residues in the wash compartment, which is probed to a waste receptacle on board the instrument or through a secondary inlet port ( not shown). In such an arrangement, the cover insertion can be, for example, semi-disposable, for example, it can be configured for replacement every 5, 10, 15, 20 or more protocols.
[00104] In a preferred embodiment, a treatment module 5000 further includes retaining means (see, for example, figure 12) configured to retain the blade 200 on the support surface 5100 during the removal of the covering component 1000, at the conclusion of a protocol. The substrate retention means can be particularly important to overcome the adhesion forces that can develop between the surface of the slide and the cover / insert element of the cover element due to the remaining reagent inside the chamber.
[00105] Advantageously, the covering component of figures 1-7 requires only two movements, while the blade is in the instrument. One movement applies the cover member to the blade and the other movement is to remove the cover member from the blade to provide access so that the treated blade can be removed and / or a new blade inserted. Minimizing the number of necessary robotics movements within an automated instrument reduces the rotation time required to complete a protocol for a given sample, as well as reducing the complexity of the instrument. In addition, in one embodiment, using positive pressure to force the reagent into the chamber, fluid distribution will be faster, as long as the waiting time for the chamber to fill under capillary action can be reduced or eliminated. The vacuum-assisted filling also increases the yield for sample processing.
[00106] Figures 8 to 15 illustrate a cover element according to another aspect of the invention. Figure 8 shows a covering element 2000, which, as a covering component 1000, has a first end 2010, the second end 2020, the first surface 2110 and the second surface 2120. An inlet 2012 is provided to the first end and an outlet 2022 is provided to the second end. Entry 2012 is in the form of a through hole (not shown), as is exit 2022. Figures 9A and 9B offer sectional end and side views, respectively, of the cover element 2000. The entry profile can vary, for example , as illustrated in figures 4a to 4e, such that multiple distributions can be received per well. Distribution nozzles do not need to contact the entrance.
[00107] An axis of rotation 2500 extends through the cover element, perpendicular to a plane that extends perpendicularly between the first and second ends. A fluid distribution end 2128 is provided, around which the cover element hinges. Figure 9c provides a bottom view of the cover member 2000, where the distribution end 2128 is visible. In use, when the cover component 2000 is in an open state, the reagent is dispensed into the entrance 2012 and drains to the interface formed by the distribution end 2128 and the blade 200. Ideally, the second surface of the control element cover 2120 and blade 200 form an angle of about 10 degrees when the reagent is dispensed into the inlet, although other angular openings are contemplated. The surface tension stabilizes the passive movement of the fluid once dispensed, while the articulation movement of the covering component 2000 around the dispensing end 2128 facilitates the movement of the reagent from the dispensing end 2128 towards the outlet 2022. Forces capillaries between the blade 200 and the cover element 2000 stabilize the fluid front as it propagates through the blade reducing the formation of bubbles.
[00108] Like the covering element 1000, the covering element 2000 provides an empty space 2124 defined by the empty boundary 2122 which presents contour walls 2126 towards the second end of the covering element. The 2126 contour walls improve the filling and emptying performance of the chamber. Figure 9c shows the entry opening 2012 for the empty space 2124. The large opening helps to prevent the formation of bubbles, which block the flow of fluid and can adversely affect the color of the sample. The area of the second surface 2120 around the empty boundary 2122 forms a locking face which, when in the closed condition, constitutes a sealing face 2130. In the closed condition, the cover member 2000 and the blade 200 are typically tightened together during an incubation period. In this condition, the reagent can also be removed by applying a vacuum to the outlet.
[00109] In color sampling systems of the prior art, a common problem has been the collection of debris and residual reagent at the limit of the chamber formed along the sealing face. Contour limiting walls 1126 in the present invention guide the reagent in the direction of exit 2022 reducing the collection of debris. It should be understood that, although exit 2022 is shown leaning against the wall of empty space 2122, this contact is not essential. Instead, the outlet opening for the void can be arranged more centrally of the cover element, such that its opening into the void is not aligned with the wall of the void.
[00110] The covering component 2000, figures 8 and 9a-9c has a shoulder 2600 that provides a surface for engaging a torsion spring of a treatment module with which the covering element can be used. The torsion spring ensures the correct angle of inclination of the covering component. This is further described in relation to figure 13.
[00111] Now going back to figures 10a-10d, a simplified version of the cover element 2000 is shown in several provisions related to blade 200. In figure 10a, blade 200 and cover element 2000 are positioned in an open condition, where the cover element 2000 is inclined with the distribution edge 2128 in contact with the slide 200. An aliquot of the reagent 300 is distributed into the entrance 2012 and the cover element 2000 is gradually pivoted towards the closed position, in a direction P, causing the distributed reagent 300 to be propagated through the slide, as shown in figure 10b. Preferably, the rate of articulation of the cover element 2000 is actively controlled according to the flow properties of the reagent. The active control of the articulation rate takes advantage of the capillarity forces between the blade 200 and the covering component 2000. Ideally, when the covering element 2000 is in the closed position (figure 10c), the reagent is distributed throughout the entire slide surface, or at least the entire sample surface. By actively moving the reagent using capillary action, the risk of bubbles forming inside the chamber is minimized.
[00112] In a preferred embodiment, the articulation action of the covering element is controlled by a 7060 controller of an automatic sample processing instrument. Typically, the controller has access to a 7126 database of pre-programmed articulation actions that increase or optimize the flow of reagent through slide 200, for a plurality of different types of reagents and / or protocols that employ various types of reagents . In some of these protocols, the 7060 controller can also be programmed to agitate the reagent by the slight movement of the cover component 2000. Alternatively, in addition, the controller can operate a vacuum pump coupled to the cover element outlet 2020, to apply a vacuum that draws the reagent through the chamber or that evacuates reagent from the chamber while the cover element is in the closed condition. The vacuum pump can also be operated in a way that causes fluid to stir inside the chamber.
[00113] Figures 10d and 12 show the covering element in a release condition where the covering component 2000 is separated (ie dissociated) from the blade 200. In this condition, a robotic arm of the instrument can load or unload a blade 200 in a 5000 treatment module, or the cover component 2000 can be cleaned, removed or replaced. The cleaning of the covering element in the release condition allows the entire second surface 2120 to be cleaned, including the walls 2122 of the void and the ceiling and the sealing surface of the covering element 2130 that contacts the blade 200. This improves methods which involve cleaning the cover element, while in the closed configuration, for example, by washing, as the debris from other reagents can remain along the "tracks" that form the sealing interface between the blade 200 and the component of cover 2000. In a preferred embodiment, cleaning of the cover element in the release state is automated by the sample processing instrument eliminating the time-consuming step of manually removing and cleaning the cover elements before loading them back onto the instrument. . The washing reagent can be drained from the cover element and into a waste receptacle on board the instrument, treated if dangerous, and in some embodiments it can be recycled.
[00114] Now, with reference to figure 11, a schematic illustration of a roofing component 2000 is shown featuring the 2012 inlet, the 2012 outlet with the 2024 pipe attached, which, in a preferred embodiment, is probed for a waste receptacle . The reagent is dispensed, typically by a robotic arm, such as FTP or a BFR at the entrance and which moves in the direction of the arrow F towards the exit. Pins 2550 arranged on the first pair of surfaces 2110 couple the cover component 2000, with articulated arms 5200 (see also figures 13 to 15).
[00115] Figure 12 is a schematic illustration of the elements of a treatment module 5000 for the treatment of a biological sample, for example, for histological staining, PCR or similar. The cover element 2000 is provided in the closed condition on a blade 200 that has a unique identification region 210 that bears a bar code. Ideally, treatment module 5000 is incorporated into an automatic sample processing instrument 7000 with a 7068 reader for reading the unique identifier and associating it with a treatment protocol to be performed on the sample carried by slide 200. Typically , reader 7068 is in communication with a controller 7060 that has access to a database that contains protocol information 7126, such as, for example, the volume of reagent to be distributed in several stages of the protocol, the rate at which the cover element is articulated to maximize capillary action by dragging particular reagents across the entire slide, reagent incubation times and incubation temperatures, optionally, shaking requirements and the like.
[00116] In a preferred embodiment, the instrument controller 7060 controls the operation of the articulated arms 5200 to articulate the covering element 2000 around the distribution end 2128, gradually moving the covering component between the open conditions (figures 10a , 10b) and closed (figure 10c, figure 12). Ideally, the pivoting action is at a speed that optimizes the flow of reagent from the dispensing end through the sample and the slide. The exploration of the capillarity between the blade 200 and the empty ceiling 2140 reinforces this movement. The ideal rate of rotation is determined, at least in part, by the viscosity of the reagent, although internal finish, coating and / or chamber geometry may also be affected.
[00117] At the conclusion of a treatment protocol, the covering component 2000 is separated from the blade and the blade is removed from the treatment module. The separation can be achieved by hinging the cover element 2000, for the opening condition and / or by displacing the cover element from the blade 200 (or vice versa) so that they are separated in the release condition ( figure 10d, figure 13). In both cases, the residual reagent in the chamber can give rise to adhesion forces that must be overcome so that the blade and the cover element 2000 are separated. Thus, in a preferred embodiment, treatment module 5000 provides blade retention means 5400 configured to retain blade 200 on support surface 5100 during separation of cover member 2000 from the blade. In the illustrated embodiment, the blade retaining means 5400 are resilient elements inclined towards the support surface 5100 so that, when the cover element 2000 is in the closed position, a portion of the blade 200 protruding from below the element cover is maintained between the blade holding device 5400 and the support surface 5100. However, it should be understood that several alternatives are contemplated, such as, for example, blade retention between a pin or rails and the support surface, devices magnetic retention devices and the like.
[00118] Figure 13 shows a view of the blade of treatment module 5000 of figure 12. The cover element 2000 is coupled to the articulated arms 5800 by pin 2550. The torsion spring 5750 engaging the lugs 2600 of the cover element ensures proper inclination orientation, before the cover element 2000 is moved from the release condition to the open state, ready to receive a reagent. Blade holding device 5400 contacts the blade when the covering element 2000 is in the closed condition and holds the blade 200 in place by overcoming the adhesion forces that can exist when the covering element and the blade are separated at the conclusion of a protocol.
[00119] Figures 14 and 15 are schematic illustrations of elements of a treatment module according to an embodiment of the invention. A blade 200 on the support surface 5100 is housed below the cover element 2000. The support surface 5100 can incorporate locating elements, such as pins 5110, 5112 and 5114, which guide the substrate into position and / or act as reference points for loading substrates of different widths. Cover element 2000 is coupled to the hinge arm 5800 by pin 2550. At a distal end, the hinge arm 5800 contacts the opening cam 5700 that hinges the arm around a second axis 5600 to move the cover element 2000 in the direction or away from the blade 200 and is therefore operable to move the cover member in the release state, as well as the open condition, where the dispensing end 2128 of the cover member 2000 contacts the blade. Ideally, the speed profile with which the articulated arm (or other drive mechanism) moves is optimized, so that the movements during the dissociation of the blade 200 and the cover element 2000 are faster than the movements made while the distribution end of the cover component is in contact with the blade and is moving from open to closed condition. The speed is reduced when the covering element 2000 is approaching the closed condition and when it exceeds adhesion forces when opening, since it is during these movements that control is most important.
[00120] To tilt the cover element from the open to the closed condition, the opening of the meat 5700 lowers the articulated arm 5800 after the "open condition" point (usually forming about 10 degrees between the second surface of the cover element and the blade) causing the cover element to rotate on the pivot axis 2550. Simultaneous rotation of the pivot arm around the pivot axis 5600 moves the pivot axis of the cover element 2550 towards the blade, such that the cover element gradually approaches the closed condition.
[00121] Advantageously, in the embodiment illustrated in figures 14 and 15, only one axis of movement is necessary to move the cover element between the open and closed release conditions. This has the added advantage of being able to accommodate any thickness of the blade. It should be understood, however, that other mechanisms can be used, which employ an articulated arm to rotate the cover element between closed and open conditions. This can be in combination with, for example, a linear actuator to raise and lower the articulation arm in order to move the cover element between the released and open conditions. Once closed, fixation devices hold the cover element and the slide together, in the closed configuration, while the reagent is incubated. In the illustrated embodiment, the fixing devices are spring-loaded 5200, although the drive mechanism used to drive the articulated arm can also be used to actively secure the cover and blade element in the closed condition.
[00122] In one embodiment, a moisture barrier is provided (like the barrier illustrated in figure 22), for example, in the form of a flexible skirt or a vapor shroud to cover a sample on a substrate, in order to mitigate sample drying or dehydration when the chamber is open. Ideally, if the sample / reagent has been heated, it will be cooled to room temperature before opening the chamber to further minimize the risk of sample dehydration. The moisture barrier can be provided as part of the covering component 2000, or as part of the treatment module 5000.
[00123] Figures 16 to 22 are schematic representations of a cover element in accordance with yet another aspect of the present invention. The cover element includes an inlet 3012, an outlet 3022, a first end 3010 and a second end 3020. An empty space 3124 is bounded by empty walls 3122 and an outer region of the second designated surface 3130 forms a sealing face when the element cover is placed in contact with a blade 200 (figures 17 to 21). The inlet profile can vary, for example, as shown in figures 4a to 4e, such that multiple distributions can be received per well. Distribution nozzles do not need to contact the entrance.
[00124] The isometric perspective view in figure 17 shows the first surface 3110 (ie, top) of the covering component 3000, together with a blade 200 that has an identifier portion 210 for carrying a unique identifier that designates the type of sample or a required protocol, or a batch identity for the sample. As figure 17 shows, when cover element 3000 is placed on slides 200, voids 3124 form a chamber for receiving reagent distributed at inlet 3012. In figure 17 the cover element and the slides are in an open condition. Figure 18 shows the same arrangement in longitudinal cross section.
[00125] Preferably, the inlet is adapted to receive multiple distributions of a reagent, so as to form a 3018 reservoir, as shown in figure 18. In another embodiment (not shown), a reagent distribution buffer capable of storing a The plurality of distributions of individual reagents can be tightly coupled to port 3012. The dispensing plug may comprise a drum that rotates between the dispensing and waiting positions. When turned to a dispensing position, a required volume of reagent is released into the inlet and drained into the distribution channel where the fluid causes the capillary reagent to pass into the 3500 space in channel 3300. The use of a buffer in this way reduces the number of individual distributions that are required by BFR or FTP robots within an automated instrument.
[00126] Figure 19 is an enlarged sectional view showing the detail of the 3300 fluid delivery functionality according to an embodiment of the invention. During the use of the reservoir of the covering element 3000, the reagent is distributed in the inlet 3012 and kept in the reservoir 3018. Fluid in the reservoir 3018 leaves the inlet through the inlet 3014 and using the surface tension of the fluid fills the distribution channel of fluid 3300 that extends over the entire width of the blade 200, as illustrated in figure 16. In the embodiment illustrated in figure 19, the channel has a smooth stepped profile that increases in height towards the first end of the cover element. This allows channel 3300 to retain a volume of reagent in the 3500 space, which feeds a fluid front, as it gradually propagates through blade 200 during movement from the open to the closed condition.
[00127] In a preferred embodiment, the space 3500 has a height of approximately 2.5 mm for a chamber volume of approximately 130 pL. The stepped profile subtends angles, as shown, where α is approximately 15 degrees, β is about 60 degrees and θ is about 8 degrees. In addition, the contoured empty space limits 3126 (figure 16) preferably have a radius of about 9 mm. If an outlet 3022 is provided, an outlet opening in the void with a diameter of about 10.3 mm is considered suitable for the effective evacuation of the reagent from the chamber.
[00128] A volume of reagent retained in the 3500 space is in contact with both the distribution channel 3300 and the blade 200. The shape of the channel 3300 is contoured in such a way that the surface tension forces inside the fluid prevent there is leakage from the channel and over the slide. In a preferred embodiment, the cover component 3000 is mounted with the side walls 3400 also being part of the fluid distribution functionality. The side walls 3400 complete the space limit 3500 within which the fluid wall is formed.
[00129] The arrangement of the channel across a width of the chamber provides a structure that facilitates the distribution of the reagent through the blade 200 by the sliding movement of the blade 200 and cover element 3000 in overlapping engagement. In one embodiment, this is achieved by moving the cursor 200 in the S direction, at the same time the cover element 3000 is kept stationary, thus carrying capillary fluid in channel 3300 and space 3500 along the sliding surface in direction F Alternatively, the cover element 3000 can be moved in the M direction, while the blade 200 is kept stationary. This also has the effect of dragging the fluid in the distribution channel 3300 and space 3500 along the surface of the slide in the F direction. Thus, in one embodiment, the reagent is distributed along the surface of the slide 200 by the relative movement of the slide and cover element 3000 from an opening condition (figures 17, 18) to a closed position (figures 20, 21). This method is referred to below as an "open" distribution.
[00130] Preferably, the closing rate is actively controlled according to the flow properties of the reagent. Reagents with higher viscosity require a slower closing speed, so that the shear forces generated during closing do not exceed the capillarity / surface tension forces that keep the fluid wall within the 3500 space that feeds the fluid front as traced through slide 200. Distribution of the reagent in this way reduces the risk of bubbles forming inside chamber 3124.
[00131] In a preferred embodiment, the sliding action of the covering element 3000 and / or blade 200 is controlled by a controller 7060 of an automated sample processing instrument 7000 of the type previously discussed. Typically, the controller has access to a 7126 database of pre-programmed slip profiles corresponding to various reagents used in the protocols performed by the instrument. Thus, the 7060 controller is configured to control the operation of a driver that optimizes the flow of reagent across the surface of the slide. An example of such a controller is shown in figure 25. In some protocols, the controller can also be programmed to agitate the reagent, causing small movements of the cover element or the blade, while in the closed condition.
[00132] The 7060 controller is shown schematically in figure 25, and includes a 7090 processor in communication with a first 7092 memory device for storing computer program code and a second 7094 memory device for storing data generated by the 7090 processor , when implementing the computer program code, through the communications infrastructure 7096. A display interface 7098 and the corresponding display 7100 allow user interaction with the controller 7060.
[00133] The 7060 controller also includes drive modules 7102 to 7112 to control the motors, pumps, scanners / readers, heat exchangers and other 7114-7124 devices necessary for the operation of the 7000 device. Treatment protocols including staining protocols (for example, example, ordering of reagents to be distributed by the BFR 7014 and the FTP robot 7028 for the corresponding slides and incubation times) are stored in a 7126 protocol database, accessible by the 7090 processor, through the 7096 communications infrastructure, allowing the 7090 processor operates the BFR 7014 and the FTP robot 7028 to distribute reagents to substrates at the blade treatment plants at the required rate.
[00134] In another embodiment, the fluid is distributed, while the cover element 3000 and blade 200 are in a closed condition. This method will be referred to as "closed" distribution and is suitable for more aqueous fluids. The closed distribution relies on the capillary action of the fluid, and not on a spreading action caused by the movement of the slide or cover element, for the reagent to distribute on the slide.
[00135] In both methods of open or closed distribution it is necessary that the chamber formed by the covering component 3000 and the blade 200 vent to the atmosphere. In the illustrated embodiment, this ventilation is provided through outlet 3022 which can also be coupled, through a valve or solenoid (not shown) to a vacuum source to evacuate the reagent chamber. However, it should be understood that an outlet 3022 on the cover element 3000 does not need to be provided. Instead, it is possible in the closed condition to maintain an opening between the blade 200 and the second end of the covering element 3020, such that the chamber is not completely closed. The omission of outlet 3022, therefore, provides a gap between the blade and the covering element, so that the chamber ventilates directly into the atmosphere, which simplifies the design of the covering and manufacturing element, however at the expense of a vacuum coupling.
[00136] The reagent distribution step in a sample treatment protocol can be followed by suppressing a second reagent. This can be preceded by evacuation from the chamber, connecting a vacuum to outlet 3022. Evacuation is reinforced by void space limits 3126 (Figure 16), which encourage the evacuation of reagent from the sealing face 3122.
[00137] In a preferred embodiment, the covering element 3000 is provided with a moisture barrier 3900 to control or limit the evaporation of moisture from the sample, once the reagent has been distributed over slide 200 and the filter element. cover has been shifted to the open condition. An example of a moisture barrier in the form of a 3900 physical enclosure is illustrated in figure 22. Providing a cover in this way limits the amount of moisture that can dissipate from the tissue sample once the chamber has been opened. In the modality shown in figure 22, the moisture barrier 3900 is adapted to cover the entire length of the blade. However, this may not be necessary. A moisture barrier that extends only over part of a slide 200 may be sufficient to limit evaporation in a way that preserves the integrity of the sample between application of the reagent and / or before the cover is slid and dispatched for further processing.
[00138] It is desirable that the 3900 moisture barrier does not interfere with a sample on the slide. Thus, moisture barrier 3900 in figure 22 comprises a substantially rigid canopy having wall sections 3910 supporting the top of the canopy. The front section 3 92 0 can be opened or closed. It should be understood, however, that the 3900 moisture barrier does not need to be a rigid or semi-rigid structure. Instead, it can be a flexible skirt or apron that extends along the blade when in the open configuration. In addition, it should be understood that the moisture barrier may, in certain embodiments, be part of or linked to a treatment module with which cover elements are used, instead of the cover element itself. When the moisture barrier is flexible, it is desirable that it can be supported, in order to maintain an opening between the moisture barrier and the sample on the slide, so as not to contaminate or affect the sample. In yet other embodiments, the moisture barrier may be a vapor wrapper consisting of a nebulized gas or water or other appropriate fluid to maintain moisture inside the sample once it has left the chamber.
[00139] Figure 23 illustrates an example of a treatment module 5000 adapted for use with the covering element 3000, where a linear actuator 5900 slides the covering element from an open condition, on blade 2 00 and to a condition closed to distribute the reagent at port 3012 over the sample carried by the slide.
[00140] In several respects, the cover element of the present invention can be adapted to allow agitation of the liquid inside the chamber. Shaking may be desirable to encourage the movement of fluid molecules in the chamber, so that there is an effective exchange between the surface of the slide (which supports the sample to be processed) and the fluid molecules. Thus, agitation of the fluid can cause more efficient processing, even with lower volumes of reagent inside the chamber. Stirring can also increase the reaction speed for a given step in a treatment protocol, thereby reducing the response time between steps. In addition, agitation of the liquid inside the chamber can reduce the impact of bubbles by movement of the bubbles inside the chamber to ensure that the entire surface of the sample is exposed to the reagent fluid during the incubation period. Stirring can be achieved by several means, including positive and / or negative pressure applied to an inlet and / or outlet port, the introduction and / or withdrawal of fluid from an inlet and / or outlet port or other means that facilitate the flow of fluid through the inlet and / or outlet openings to generate sufficient turbulence to promote fluid agitation.
[00141] In addition, the agitation of the fluid can reduce the stain on the artifacts resulting from the presence of bubbles, increase the uniformity of reagent throughout the chamber, minimization of "dead zones", facilitating cleaning and / or washing in situ of the surface cover element. The movement of the fluid inside the chamber can be enhanced by a contour geometry of the walls of the chamber (for example, as described with reference to figure 1). Contours can be provided at an inlet end of the cover element to improve fluid flow and, when supplied to the outlet end, can provide better fluid evacuation from the chamber, such that little debris remains. Alternatively and in addition, one or both ends of the chamber can be defined by walls that form a curved or tapered end wall. In addition, it may be desirable to provide a coating on the empty ceiling that increases the spread of reagent within the chamber. The coating may consist of a texture, such as, for example, wrinkling, engraving, ripples or arrow outlines on the second surface forming the empty space ceiling. The slope of the ceiling of empty space or curve of it or provision of undulations within the ceiling can also improve the flow of fluid. Alternatively and additionally, the ceiling and / or walls of the void can be coated or treated with a coating material that improves fluid flow.
[00142] Several aspects of the present invention give rise to cover elements that minimize the amount of reagent needed to perform the steps of the treatment protocol of the type employed by the 7000 instrument. Ideally, several aspects of the present invention facilitate a reaction chamber effective formed by the covering element, which has a volume as small as 120 to 135 pL. Although volumes as small as 30 pL are also contemplated. In some reactions, it may be necessary to provide a larger reaction chamber with closed volumes of, for example, up to 200 pL.
[00143] In one or more modalities, the detection of the liquid level for the reagents distributed at the entrance may be desirable. Detection of the liquid level can be performed using touch probe technology and / or can be performed by monitoring changes in capacitance or pressure at the end of a distribution probe. Alternatively, optical liquid level detection systems and ultrasound systems can be employed. Measurements of reagent volumes made at the entrance, in the chamber and / or through the exit can be compared by a controller 706 0 on the edge of an automated instrument 7000 for cross checking against the volume of distribution calculated according to the number of protocols performed. This cross-check can then be used to control the inventory of reagents stored on the edge of the automated instrument.
[00144] Although the various modalities of the cover element illustrated demonstrate only one outlet, it should be understood that several outlets could be provided. However, when a vacuum is applied to increase the movement of liquids (including agitation) inside the chamber and / or evacuation of the reagent from the chamber, separate vacuum sources are required for each of the outlets. Thus, when designing a cover element according to the present invention, the person skilled in the art will balance complexity and price with performance. Although each or more of the outlets can be coupled to a vacuum source, modalities that use forced pressure distribution (figures 1 to 3) and capillary distribution, requiring that the chamber be ventilated to the atmosphere. Therefore, the outlet can be interchangeably coupled with a vacuum source and an opening for the atmosphere.
[00145] The use of a vacuum when filling the chamber can reduce the likelihood of bubbles forming inside the chamber. In addition, the use of a vacuum to evacuate fluid from the chamber reduces the likelihood of residues remaining within the chamber between reagent distributions. Another advantage of using a vacuum to remove the reagent from the chamber is that less reagent can be used, since evacuation of the chamber, before the application of the second reagent, minimizes the risk of mixing.
[00146] Ideally, the 7060 automated controller instrument accesses a 7128 database of protocol information, which is used to control one or more vacuum sources to apply the correct vacuum magnitude and duration, depending on the reagent used (eg example, viscous or aqueous) and / or the type of sample or cut thickness (for example, skin sample or cytological sample which can vary in thickness from 1 pm to 15 pm and preferably from 3 pm to 5 pm).
[00147] It should be noted that the entrance can be formed on the cover element body in any orientation, and can leave the covering element on the entire surface, although in the illustrated modalities the entrance opening is provided on the first surface (ie , top) of the cover element. In addition, it should be noted that, although each modality is illustrated with an entry, the provision of multiple entries is also contemplated. Likewise, as described above, several exits are contemplated. It should be understood that these exits can leave the covering element on the entire surface, although the illustrated modalities show the exit leaving the covering element on the first surface (that is, on the top) and the front surface (figure 1). The location of the outlet opening can be influenced by the location of the couplings and / or conductors that connect the outlet to a vacuum source and / or a valve or solenoid through which the outlet is coupled with the vacuum source and / or ventilated to the atmosphere. In addition, although not essential, the location of the outlet in contact with a limit of empty space inside the cover element can improve the evacuation of reagent from the chamber.
[00148] Throughout this specification, the illustrated modalities are described with reference to the blade being maintained with a substantially horizontal orientation. It should be understood, however, that horizontal orientation is not necessarily required and that the support surface can support the blade at an angle. In addition, the invention is described in terms of fluid transmission longitudinally, from the first end to the second end of the cover element. It should be understood, however, that the cover element can be configured for the flow of transverse fluid across the blade using a wider fluid front, although the risk of bubble formation may be higher in this configuration. It is also noted that the processing of the blade according to embodiments of the invention need not be limited to processing a horizontal orientation.
[00149] Preferably, when the cover element of the invention is used by an automated sample processing instrument, each slide to be processed contains a unique identifier, such as a barcode or RFID tag that identifies one or more among types sample and a protocol to be performed on the sample. This information is detected by a reading device on the instrument and used to program actions for the distribution of BFR and FTP robots within the instrument, according to the required protocol.
[00150] Whenever the terms "comprises", "comprise", "comprised" or "comprising" are used in this specification (including the claims), they are interpreted as indicating the presence of the indicated characteristics, integers, steps or components, but not preventing the presence of one or more other characteristics, whole numbers, steps or components or groups thereof.
[00151] It should be understood that various modifications, additions and / or changes can be made to the parts previously described without departing from the scope of the present invention as defined in the appended claims.

权利要求:
Claims (35)
[0001]
1. Module for the treatment of a biological sample, the module characterized by the fact that it comprises: a. a cover element comprising i. opposite ends comprising a first end and a second end; ii. opposite surfaces comprising a first surface and a second surface; iii. a void on the second surface which, when juxtaposed with a substrate, forms a chamber; and iv. a fluid inlet towards the first end and in fluid communication with the empty space; a support surface for a substrate with a biological sample on it; and operable fixing device to retain the removable cover element in juxtaposition with the substrate for an incubation period, and a pivot arm configured to rotate the cover element around a dispersing edge, causing the fluid to move from the entrance along the substrate of the spreading edge towards a fluid outlet, the articulation arm being operable to rotate the cover element to an open condition and to a closed condition, in which the articulation arm is operable to rotate the cover element and raise the cover element from the support surface, so that, from the first and second opposite ends, only one of the first and second opposite ends is lifted from the support surface.
[0002]
2. Treatment module, according to claim 1, characterized by the fact that it comprises a robot configured to position one or both the substrate and the covering element in the treatment module; and optionally in which the robot is configured to dispense a reagent for admission of the cover element during a treatment protocol.
[0003]
Treatment module according to claim 1 or 2, characterized in that it comprises an operable coupling to alternately connect one or more outlets, including the outlet fluid of the cover element with one or more fans to the atmosphere and respective one or more sources of negative pressure.
[0004]
Treatment module according to claim 1, characterized in that it additionally comprises a substrate retaining member configured to retain the substrate on the support surface during the opening of the chamber or separation of the covering element and the substrate.
[0005]
5. Treatment module, according to claim 1, characterized by the fact that the treatment module is configured for use with an automated sample processing instrument comprising a plurality of treatment modules operable independently under the control of a controller. instrument; and in which the operation of one or more of a clamp, a heat exchanger, a robot, a source of negative pressure and a fluid dispenser is under the control of the instrument controller.
[0006]
Treatment module according to claim 1, characterized in that it also includes a washing compartment for exposing a surface of the cover element to a washing reagent, where the support surface is optionally molded to receive a substrate with a sample in it and, in the absence of a substrate, to form the wash compartment.
[0007]
7. Method for the incubation of a biological sample with one or more reagents using a covering element as defined in claim 1, characterized by the fact that it includes the steps of: a. provision of the sample on a substrate; B. positioning the substrate and the covering element to form the chamber; ç. positioning a tip of the distribution probe in combination contact with a fluid inlet of the cover element; and d. directing a first volume of a first reagent into the inlet, with sufficient force for the first reagent to substantially cover the sample on the substrate; and optionally shaking the reagent chamber; and stirring the reagent inside the chamber.
[0008]
Method according to claim 7, including one or more additional steps, characterized in that it comprises: a. apply negative pressure to a fluid outlet of the cover element to aspirate reagent into the chamber towards the outlet; and b. remove the substrate from a support surface and immerse a second surface of the cover element in a washing reagent.
[0009]
9. Method for incubating a biological sample with one or more reagents using a treatment module as defined in claim 1, characterized in that it includes the steps of: a. provide the sample on a substrate; B. positioning the substrate and a covering element to form a chamber; ç. positioning a dispensing probe tip in contact with a fluid inlet of the cover element; and d. conducting a first volume of a first reagent into the inlet with sufficient force that the first reagent substantially covers the sample on the substrate; and optionally and stir the reagent inside the chamber.
[0010]
Method according to claim 7, characterized in that it includes one or more additional steps, comprising: a. apply negative pressure to the outlet of the cover element to aspirate the reagent into the chamber towards the outlet and b. remove the slide from a support surface of the treatment module and immerse a second surface of the cover element in a washing reagent.
[0011]
11. Treatment module, according to claim 1, characterized by the fact that the cover element is removable from the treatment module.
[0012]
12. Treatment module according to claim 1, characterized by the fact that the empty cover element is limited by empty walls having one or more contoured regions to improve the movement of the fluid within the chamber.
[0013]
13. Treatment module, according to claim 1, characterized by the fact that the covering element includes: the outlet of fluid towards the second end and in fluid communication with the void; and a guide at the entrance, configured to direct fluid to the entrance.
[0014]
14. Treatment module according to claim 1, characterized by the fact that the cover element includes: the fluid outlet, the fluid outlet being disposed towards the second end and in fluid communication with the void; and the dispersing edge, the dispersing edge being arranged in fluid communication with the fluid inlet; wherein the cover member is adapted to rotate around the dispersing edge, causing movement of the fluid in the fluid inlet from the dispersing edge towards the fluid outlet.
[0015]
15. Treatment module, according to claim 1, characterized by the fact that the cover element includes a fluid dispersion feature from which the fluid is dispensed, in which the fluid dispersion feature is configured to dispense fluid from the inlet to at least one width of the substrate.
[0016]
16. Treatment module, according to claim 1, characterized by the fact that the cover element further comprises the outlet of fluid towards the second end and in fluid communication with the void and through which the fluid can be withdrawn .
[0017]
17. Treatment module, according to claim 1, characterized by the fact that the empty cover element is bounded by empty walls with one or more contoured regions and one or more contoured regions comprise rounded corners connecting side walls of the void with a end wall of the cover element to encourage fluid flow into the chamber.
[0018]
18. Treatment module, according to claim 1, characterized by the fact that the empty cover element is bounded by empty walls having one or more contoured regions and in which one or more contoured regions comprise a rounded cone that connects the side walls of the void with an end wall of the cover element to encourage fluid flow within the chamber.
[0019]
19. Treatment module, according to claim 1, characterized by the fact that the empty cover element is bounded by empty walls with one or more contoured regions and in which one or more contoured regions comprise rounded cornices that connect the walls of the void with an empty roof on the second surface of the cover element.
[0020]
20. Treatment module according to claim 1, characterized by the fact that an empty roof on the second surface of the cover element has a finish configured to improve the spread of reagent from the fluid inlet to the fluid outlet and, optionally , where the finish is at least one of engraved, wavy, wavy, inclined, curved and wavy.
[0021]
21. Treatment module, according to claim 1, characterized by the fact that the cover element is disposable.
[0022]
22. Treatment module according to claim 1, characterized in that the cover element is formed from at least two parts, including a cover element body and a cover element insert, wherein the cover element cover element insertion is configured to form the chamber with the substrate; and optionally, wherein the insertion of the cover element is disposable.
[0023]
23. Treatment module according to claim 1, characterized by including a guide configured to form a comfortable fit with a distribution probe tip.
[0024]
24. Treatment module, according to claim 1, characterized by the fact that the guide has sufficient conformity to receive and form a seal with a distribution probe tip.
[0025]
25. Treatment module for a biological sample, the module characterized by the fact that it comprises: a covering element having; i. opposite ends comprising a first end and a second end; ii. opposite surfaces comprising a first surface and a second surface; iii. a void on the second surface which, when juxtaposed with a substrate, forms a chamber; and iv. a fluid inlet towards the first end and in fluid communication with the void; a support surface for a substrate containing a biological sample; a pivot arm attached to the cover element and located at the first or second end thereof to position the cover element in an open condition and in a closed condition, the closed condition being a position where the cover element and the substrate are juxtaposed to form the chamber, the actuation arm being connected to a clamp that resiliently tilts the cover element in juxtaposition with the substrate for an incubation period; and a torsion spring attached to the cover element of the drive arm and configured to exert a force on the cover element towards the substrate, and in which the fluid inlet extends through the cover element.
[0026]
26. Treatment module according to claim 1, characterized by the fact that the articulation arm comprises pins extending from the articulation arm and coupling the cover element to the articulation arm.
[0027]
27. Treatment module according to claim 1, characterized in that the articulation arm is configured in such a way that turning the cover element from the closed condition to the open condition raises the first opposite end of the cover element from the substrate without lifting the second opposite end of the substrate cover element, where the first end of the cover element comprises the fluid outlet, and the second end of the cover element comprises the fluid inlet and the spreading edge.
[0028]
28. Treatment module according to claim 1, characterized by the fact that the covering element comprises the spreading edge at the fluid inlet, and in which the fluid inlet comprises a step that increases in height, along a longitudinal axis of the fluid inlet through the cover member and narrows the fluid inlet towards the dispersing edge of the fluid inlet.
[0029]
29. Treatment module according to claim 28, characterized in that it further comprises the substrate, and in which, in the closed condition in which the covering element is sealed to the substrate, a lower portion of the step on the dispersing edge forms an angle of 15 ° to the substrate and an upper portion of the step away from the shape of the spreading edge an angle of 8 ° to the substrate, and the step comprising an angle of 60 ° from the lower portion to the upper portion in relation to the substrate.
[0030]
30. Treatment module, according to claim 1, characterized by the fact that it also includes: a controller configured to implement the control of the articulation arm: to rotate the cover element around the spreading edge, rotate the cover element to the open condition and the closed condition, and to rotate the cover element from the closed condition to the open condition by lifting at least part of the cover element from the substrate.
[0031]
31. Treatment module according to claim 25, characterized in that the fluid inlet extends through the covering element from the first surface to the second surface.
[0032]
32. Treatment module according to claim 1, characterized by the fact that the articulation arm is still operable to lift the covering element of the substrate while maintaining contact between the spreading edge and the substrate, rotating the covering element to the along a longitudinal axis of the covering element in relation to a longitudinal axis of the substrate.
[0033]
33. Treatment node according to claim 32, characterized in that the spreading edge of the covering element is on a longitudinal end side of the covering element.
[0034]
34. Treatment module according to claim 28, characterized in that the longitudinal axis of the fluid inlet is perpendicular to the longitudinal axis of the cover element, and in which the fluid inlet gradually narrows from part of the step , farther from the substrate, to the dispersion edge.
[0035]
35. Treatment module according to claim 31, characterized in that the torsion spring extends from the cover element to at least one of the pins.

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JP2018021932A|2018-02-08|

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US20140315256A1|2014-10-23|

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EP2780721A4|2015-07-01|

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IN2014CN04294A|2015-09-04|

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BR112014011935A2|2017-05-30|

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AU2012339616A1|2014-06-12|

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CA2855511A1|2013-05-23|

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EP2780721A1|2014-09-24|

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CN104094122B|2017-03-22|

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法律状态:
2018-12-04| B06F| Objections, documents and/or translations needed after an examination request according [chapter 6.6 patent gazette]|

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2019-12-17| B06U| Preliminary requirement: requests with searches performed by other patent offices: procedure suspended [chapter 6.21 patent gazette]|

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2020-06-16| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|

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2020-11-03| B16A| Patent or certificate of addition of invention granted|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 15/11/2012, OBSERVADAS AS CONDICOES LEGAIS. |

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PCT/AU2012/001407|WO2013071352A1|2011-11-16|2012-11-15|Cover member, method and treatment module for treating a biological sample on a substrate|

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