EP4680932A1

EP4680932A1 - Automated staining apparatus with improved staining slot module

Info

Publication number: EP4680932A1
Application number: EP23822481.0A
Authority: EP
Inventors: Ivans GERCEVS; Artur MEZHEYEUSKI; Patrick MICKE; Carina STRELL; Taisija KOZARINA; Andris ZACS; Irina PTICINA; Vladimirs Petrovs
Original assignee: Argento Lab Sia; Transporta Un Sakaru Instituts AS
Current assignee: Argento Lab Sia; Transporta Un Sakaru Instituts AS
Priority date: 2023-03-15
Filing date: 2023-12-04
Publication date: 2026-01-21
Also published as: WO2024191277A1

Abstract

The present invention relates to automated staining apparatuses and especially to staining slot modules of the automated staining apparatuses. The staining slot module comprises a base plate, a board arranged on the base plate. A slide is arrangeable on the same base plate. A chip is arrangeable within the base plate and the board and on the slide. The staining slot module is characterized in that it comprises a pressing mechanism allowing secure and adjustable positioning of the chip in the staining slot module.

Description

AUTOMATED STAINING APPARATUS WITH IMPROVED STAINING SLOT MODULE

DESCRIPTION

Field of the invention

[001] The present invention relates to automated staining apparatuses and especially to staining slot modules of the automated staining apparatuses.

Background of the invention

[002] In research and medicine different methods of tissue staining are used, to reveal the structure of the tissue due to the unequal chemical affinity of various tissue elements for histological dyes. The most commonly used method is the staining with haematoxilin and eosin (H&E staining).

[003] The methods of immunostaining or immunohistochemical staining (IHC) and in-situ hybridization (ISH) are used for localizing the proteins or the mRNA and gene expression within the human or animal tissue samples. To visualize (to label) the presence of proteins, mRNA and genes different types of dyes or markers are used, which could be generally separated into two major classes: chromogenic and fluorescent ones. The procedures of IHC and ISH are therefore completed by the visualization stage, and thus will be denoted here by the term ‘staining’.

[004] Before staining, the tissue section is mounted on a special glass-slide. Staining, like all other stages of the process of preparing a histological specimen, can be performed manually and automatically (with varying degrees of automation).

[005] Modern applications require the IHC and ISH and other biological tissue staining methods, for the detection and visualization of several different specific targets in the same tissue sample (protein, RNA or gene): multiplex IHC, multiplex ISH, etc. Due to the specifics of the staining process, in most cases, multiplex staining requires multiple iterative replies to the staining protocol. [006] Manual method for IHC or ISH procedure requires a lot of time (up to several days) and a qualified specialist; all processes are performed manually, using different systems and equipment for staining. Automatic method for IHC or ISH procedure is performed by special systems - auto-stainers with varying levels of automation. The use of staining systems reduces the workload of qualified personnel and significantly increases productivity, as well as (in the ideal case) standardization of the process. Large medical research centers and institutions working with histological studies use automatic staining systems.

[007] International application publication No. WO02/072264 discloses a system for providing a fluidic interface to slides bearing microarrays of biomolecules or other samples immobilized thereon to perform a variety of chemical reactions or processing steps on the slide. An interface device seals against the slide to form a chamber or chambers containing all or a portion of the microarray, providing selective access to portions of the slide. The interface device includes inlet and outlet ports permitting liquid sample and reagents to be introduced to and removed from the chamber accessing the slide surface. Pre- and post- array microfluidic circuitry may be included in the interface device or in attachable modules. Similar design is disclosed in German patent application publication No. DE 103 16 723.

[008] International application publication No. WO2013/128322 discloses a biological and chemical sample processing slot module that comprises a high pressure-resistant, shallow and wide area microfluidic chamber having at least one wall formed by a detachable slide containing samples such as immobilized entities, biological samples or molecules. The slot module comprises an arrangement of microfluidic access holes for injecting to and collecting fluid from said chamber. The slot module is interfaced with inlet ports and microfluidic channels which are formed external to the chamber. Similar design is disclosed in the international application publication No. W003/106033.

[009] U.S. patent publication No. 5,958,760 discloses a support retaining member for use in processing a sample supported on a support. The member being such than when assembled together with a support it forms a support cell. The support cell comprising a substantially sealed chamber, and the chamber being provided with a fluid inlet and a fluid outlet for the introduction and removal respectively of fluids used in processing the sample. [010] US patent application publication No. US2001/003652 discloses a staining system wherein the slides are securely held by two pressing screws, on each side. US patent application publication No. US2022/062898 discloses a staining device wherein the slide is pressed and secured in place by means of four release coupling mechanisms. US patent application publication No. US2022/291246 discloses a staining module wherein the alternative securing mechanism comprises pivotable cover pressing plate which is fixedly secured by actuation of two sliding locking rods. US patent application publication No. JP6688323 discloses a staining module wherein the alternative securing mechanism comprises a pivotable cover pressing plate which is actuated by a linear actuator assembly.

Summary of the invention

[Oi l] The present invention is a staining slot module for auto-stainer system. The invention is set out in the appended set of claims. The staining slot module comprises a base plate, a board arranged on the base plate, a slide arranged on the same base plate, a chip positioned within the base plate and board and on the slide. The staining slot module is characterized in that it comprises a pressing mechanism allowing secure and adjustable positioning of the insertable chip and slide in the staining slot module.

[012] The base plate has a top surface and a lower surface and a base plate recess in the middle section thereof. The base plate may be considered as the main element of the staining slot module as other elements of the module are associated with the base plate.

[013] The board has an open cut-out which wider than the base plate recess. The open cut-out is arranged around the base plate recess so as to form a shoulder. The form of the open cut-out is such that the board resembles U-type part.

[014] In other embodiments of the invention the base plate and the board may be an integral part.

[015] The slide is formed so that the slide is slidable into the open cut-out of the board and supported by the base plate. Later the chip is based on the slide. [016] The chip has an upper surface and a lower surface. The chip comprises an open-type chamber formed in the lower surface of the chip. The chip also comprises an inlet hole and an outlet hole, both formed on the lower surface of the chip. The inlet hole is fluidly connected to the open-type chamber via an inflow channel. The open-type chamber is fluidly connected to the outlet hole via an outflow channel. Aforementioned communication provides staining liquid flow in/out from the chip. The chip further comprises a truncated edge merged with the open-type chamber. The inflow channel enters the open-type chamber at the truncated edge of the chip.

[017] The staining liquid flow is further provided by a port attached to the lower surface of the base plate and the port comprises an inlet aligned with an inlet hole of the chip and an outlet aligned with an outlet hole of the chip. This connection is provided through the inlet and outlet holes of the base plate.

[018] The staining slot module further comprises a heat transfer unit attached to the lower surface of the base plate for providing preset temperature or heat to the chip.

[019] The staining slot module comprises a pressing plate having a top surface and a lower surface. The pressing plate is arranged above and pressed against the chip. The pressing plate is associated with the pressing mechanism for providing a predictable and firm pressing force of the pressing plate against the chip. The pressing mechanism comprises a rod having first end and the second end. The rod is arranged through the holes or openings in the pressing plate and the base plate so that the first end of the rod extends above the top surface of the pressing plate and the second end of the rod extends beyond the lower surface of the base plate. The second end of the rod is threaded. The pressing mechanism comprises connection nuts fixing the rod to the base plate. The pressing mechanism comprises a spring arranged on the part of the rod that extends beyond the lower surface of the base plate, and a thrust nut mounted on the second end of the rod so as to pretension the spring according to the necessary pressing force. The spring force is adjusted by repositioning of the thrust nut onto the rod. The pressing mechanism further comprises an eccentric lock mounted onto the first end of the rod so as to provide adjustable movement of the pressing plate in relation to the chip securing the chip against the slide and the base plate. The pressing mechanism of the present invention provides easy and pre-settable securing of the chip in the staining slot module, as well as fast removal or change of chips within the staining slot module. [020] In other embodiments of the invention the base plate and the board may be an integral part.

Brief description of the drawings

[021] The drawings illustrate generally, by way of example, but not by way of limitation, various embodiments of the invention.

[022] Fig. l is a perspective view of an auto-stainer with four staining slot modules.

[023] Fig. 2A is a top perspective view of a staining chip with visible hidden lines.

[024] Fig. 2B is a bottom view of the staining chip with visible hidden lines.

[025] Fig. 2C is a side view of the staining chip with visible hidden lines.

[026] Fig. 2D is an A-A cross-section view of the staining chip as seen in Fig 2C.

[027] Fig. 2E is a B-B cross-section view of the staining chip as seen in Fig 2C.

[028] Fig. 2F is a C-C cross-section view of the staining chip as seen in Fig 2C.

[029] Fig. 3 is an exploded view of a staining slot module.

[030] Fig. 4 is another exploded view of the staining slot module as seen in Fig. 3.

[031] Fig. 4A is a top view of the staining slot module.

[032] Fig. 4B is an A-A cross-section view of the staining slot module as seen in Fig. 4A.

[033] Fig. 4C is a B-B cross-section view of the staining slot module as seen in Fig. 4A.

[034] Fig.4D is a C-C cross-section view of the staining slot module as seen in Fig. 4A. [035] Fig. 5 A is another top view of the staining slot module.

[036] Fig. 5B is a D-D cross-section view of the staining slot module as seen in Fig. 5A.

[037] Fig. 5C is an E-E cross-section view of the staining slot module as seen in Fig. 5A.

[038] Fig. 6A is another top view of the staining slot module.

[039] Fig. 6B is a G-G cross-section view of the staining slot module as seen in Fig. 6A.

[040] Fig. 7 is a top perspective view of a base plate (3) and a board (4).

[041] Fig. 8 is a bottom perspective view of a pressing plate (9).

Detailed description of the embodiments

[042] The preferred embodiments of the invention are now described with reference to the figures to illustrate the objectives, the advantages, and efficiency of the present invention.

[043] Fig. 1 is a perspective view of an auto-stainer (1) with four staining slot modules (2). In other embodiments of the invention, the auto-stainer (1) may comprise more than four staining slot modules (2) as the auto-stainer (1) is easily scalable. The auto-stainer (1) also comprises a module for storage and management of staining liquids (100), in which test-tube cartridges (101) are arranged.

[044] Figs. 2A to 2F illustrate a chip (6) also known as a staining chip. The chip (6) has an upper surface (68) and a lower surface (69). The chip (6) further comprises an open-type chamber (63) formed in the lower surface (69) of the chip (6). The chip (6) comprises an inlet hole (61) and an inflow channel (64). The inflow channel (64) provides fluid connection between the inlet hole (61) and the open-type chamber (63) so that the staining liquids can be supplied to the open-type chamber (63) of the chip (6). The chip (6) comprises an outlet hole (62) and an outflow channel (65). The outflow channel (65) provide fluid connection between the open-type chamber (63) and the outlet hole (62) so that the staining liquid can be drained from the open-type chamber (63). The chip (6) further comprises a truncated edge (66) merged with the open-type chamber (63) of the chip (6). The inflow channel (64) enters the open-type chamber (63) at the truncated edge (66) of the chip (6).

[045] Figs. 3 to 6B illustrate one embodiment of the staining slot module (2) in more detail. The staining slot module (2) comprises a base plate (3) having a top surface (31) and a lower surface (32) and a base plate recess (33) in the middle section thereof. The staining slot module

(2) also comprises a board (4) having an open cut-out (41) so that the board (4) has an U-type profile. The open cut-out (41) is wider than the base plate recess (33) of the base plate (3) and the open cut-out (41) is arranged around the base plate recess (33) so as to form a shoulder (34). The staining slot module (2) also comprises a slide (5) formed so that the slide (5) is slidable into the open cut-out (41) of the board (4) and supported by the base plate (3). The chip (6) with its lower surface (69) is positioned on the slide (5) and the inlet hole (61) of the chip (6) is aligned with an inlet hole (37) of the base plate (3) and the outlet hole (62) of the chip (6) is aligned with an outlet hole (36) of the base plate (3). Moreover, the inlet hole (37) and the outlet hole (36) of the base plate (3) are arranged on the shoulder (34) of the base plate

(3). All features of the base plate (3) and the board (4) are illustrated in Fig. 7.

[046] The staining slot module (2) comprises a port (7) attached to the lower surface (32) of the base plate (3) and the port (7) comprises an inlet (71) aligned with an inlet hole (61) of the chip (6) and an outlet (72) aligned with an outlet hole (62) of the chip (6). The port (7) provides supply and drainage of the staining liquid. The staining slot module (2) further comprises a heat transfer unit (8) attached to the lower surface (32) of the base plate (3) (see Figs. 3 to 6B).

[047] The staining slot module (2) comprises a pressing plate (9) having a top surface (91) and a lower surface (92). The pressing plate (9) is in combination with a pressing mechanism. The pressing plate (9) is arranged above the chip (6) and is pressed against the same chip (6). Figs. 3 to 6B illustrate the pressing plate (9) in various states and in addition, all features of the pressing plate (9) are illustrated in Fig. 8. The pressing plate (9) comprises a recess (93) shaped or resembles a profile or outline of the chip (6) providing more aligned positioning of the pressing plate (9) onto the chip (6). The pressing plate (9) also comprises two rod slots (94) for two rods (11) of a pressing mechanism. [048] The pressing mechanism provides a predictable and firm pressing force of the pressing plate (9) against the chip (6). The pressing mechanism comprises a rod (11) having first end (12) and the second end (13). The rod (11) is arranged through the rod slots (94) of the pressing plate (9) and the base plate (3) so that the first end (12) of the rod (11) extends above the top surface (91) of the pressing plate (9) and the second end (13) of the rod (11) extends beyond the lower surface (32) of the base plate (3). The second end (13) of the rod (11) is threaded. The pressing mechanism comprises a spring (14) arranged on the part of the rod (11) that extends beyond the lower surface (32) of the base plate (3). The pressing mechanism also comprises a thrust nut (15) mounted on the second end (13) of the rod (11) so as to pretension the spring (14). The pressing mechanism comprises connection nuts (17) for fixing the rods (11) to the base plate (3). The pressing mechanism also comprises an eccentric lock (16) mounted onto the first end (12) of the rod (11) so as to provide pressing force against the pressing plate (9) which further presses the chip (6) against the slide (5) and the slide against the base plate (3) and locking aforementioned elements together. The spring (14) force is adjustable by repositioning of the thrust nut (15) onto the rod (11).

[049] While the invention may be susceptible to various modifications and alternative forms, specific embodiments of which have been shown by way of example in the figures and have been described in detail herein, it should be understood that the invention is not intended to be limited to the particular forms disclosed. Rather, the invention includes all modifications, equivalents, and alternatives falling within the scope of the invention as defined by the following claims.

Claims

1. A staining slot module (2), comprising: a base plate (3) having a top surface (31) and a lower surface (32) and a base plate recess (33) in the middle section thereof; a board (4) having an open cut-out (41), wherein the open cut-out (41) is wider than the base plate recess (33), and the open cut-out (41) is arranged around the base plate recess (33) so as to form a shoulder (34); a port (7) attached to the lower surface (32) of the base plate (3); a heat transfer unit (8) attached to the lower surface (32) of the base plate (3); a pressing plate (9) having a top surface (91) and a lower surface (92), and the pressing plate (9) is arranged above an insertable chip (6) and pressed against the same chip (6); and a pressing mechanism for providing a predictable and firm pressing force of the pressing plate (9) against the chip (6), wherein the pressing mechanism comprises: a rod (11) having first end (12) and the second end (13), wherein the rod (11) is arranged through the pressing plate (9) and the base plate (3) so that the first end (12) of the rod (11) extends above the top surface (91) of the pressing plate (9) and the second end (13) of the rod (11) extends beyond the lower surface (32) of the base plate (3), and wherein the second end (13) of the rod (11) is threaded; connection nuts (17) fixing the rod (11) to the base plate (3); a spring (14) arranged on the part of the rod (11) that extends beyond the lower surface (32) of the base plate (3); a thrust nut (15) mounted on the second end (13) of the rod (11) so as to pretension the spring (14); and an eccentric lock (16) mounted onto the first end (12) of the rod (11) so as to provide adjustable movement of the pressing plate (9) in relation to the chip (6) securing the chip (6) against the base plate (3).

2. The staining slot module (2) according to Claim 1, wherein the module (2) comprises a chip (6) having an upper surface (68) and a lower surface (69), wherein the chip (6) comprises: an open-type chamber (63) formed in the lower surface (69) of the chip (6); an inlet hole (61); an inflow channel (64) fluidly connecting the inlet hole (61) with the open-type chamber (63); an outlet hole (62); and an outflow channel (65) fluidly connecting the open-type chamber (63) with the outlet hole (62).

3. The staining slot module (2) according to claim 1 or 2, wherein the chip (6) further comprises a truncated edge (66) merged with the open-type chamber (63) of the chip (6), and wherein the inflow channel (64) enters the open-type chamber (63) at the truncated edge (66) of the chip (6).

4. The staining slot module (2) according to any of claims 1 to 3, wherein the spring (14) force is adjusted by repositioning of the thrust nut (15) onto the rod (11).

5. The staining slot module (2) according to any of claims 1 to 4, wherein the base plate (3) and the board (4) are an integral part.

6. The staining slot module (2) according to any of claims 1 to 5, wherein the module (2) comprises a slide (5) formed so that the slide (5) is slidable into the open cut-out (41) of the board (4) and supported by the base plate (3).

7. The staining slot module (2) according to any of claims 2 to 6, wherein the port (7) comprises an inlet (71) aligned with the inlet hole (61) of the chip (6) and an outlet (72) aligned with the outlet hole (62) of the chip (6).