A fully automatic immunohistochemical staining device

Abstract

The present invention discloses a fully automatic immunohistochemical staining device, which includes a micro reagent sampling mechanism and a plurality of incubation units. The incubation unit includes a heating mechanism, a large reagent sampling mechanism, a cleaning mechanism, a waste suction mechanism, a coverslip conversion mechanism and a transverse movement mechanism. The transverse movement mechanism can drive the sampling port of the large reagent sampling mechanism, the liquid inlet of the cleaning mechanism and the waste suction port of the waste suction mechanism to move; the coverslip conversion mechanism includes a two-axis movement mechanism, a coverslip support and a rotation conversion mechanism. The coverslip support is respectively provided with a large reagent coverslip installation position and a micro reagent coverslip installation position. The coverslip support is movably installed on the two-axis movement mechanism. The two-axis movement mechanism can drive the coverslip support to move along a straight line, and the rotation conversion mechanism drives the coverslip support to rotate. The micro reagent sampling mechanism includes a three-axis movement mechanism and a sampling needle installed thereon. The three-axis movement mechanism drives the sampling needle to move. This device has the advantages of high throughput processing capacity, extremely high reagent utilization rate and less cross contamination, etc.

Description

Technical Field

[0001] The present invention relates to the technical field of immunohistochemistry equipment, and particularly relates to a fully automatic immunohistochemistry staining device. Background Art

[0002] Immunohistochemistry technology has the advantages of high specificity, strong sensitivity and simple operation, which makes immunohistochemistry technology widely promoted and applied in the field of disease diagnosis, especially in clinical pathological diagnosis and tumor transformation diagnosis. Immunohistochemistry technology is widely used in clinical pathological diagnosis and tumor transformation diagnosis, and its test results are related to the physical and mental health of the tested personnel. The safety, rigor and accuracy of its detection are particularly important. There are many factors affecting immunohistochemistry staining, including environmental conditions, tissue processing, antibody quality, staining process, etc. Traditional manual operations have many steps and take a long time, and it is inevitable to produce artificial experimental errors and staining quality problems during the operation process, which affect the accuracy and stability of the staining results and even mislead pathologists.

[0003] Currently, the immunohistochemistry equipment on the market generally includes brands such as Roche, Leica, and Dako. However, these equipments have problems such as insufficient capacity of the glass slides and relatively long reaction time. Moreover, due to being foreign brands, the equipment procurement cost and technical maintenance cost are quite high. Summary of the Invention

[0004] In order to solve the problems existing in the above-mentioned prior art, the present invention provides a fully automatic immunohistochemistry staining device, which has a relatively simple structure, and has the advantages of high-throughput staining processing ability, extremely high reagent utilization rate and less cross-contamination.

[0005] The technical solution adopted to achieve the above object of the present invention is as follows:

[0006] A fully automatic immunohistochemistry staining device includes a frame, an incubation unit and a micro reagent injection mechanism. There are multiple incubation units, and the multiple incubation units are sequentially fixed on the frame along a straight line;

[0007] The incubation unit includes a heating mechanism, a glass slide module, a large reagent addition mechanism, a cleaning mechanism, a waste suction mechanism, a coverslip conversion mechanism, a transverse movement mechanism and an incubation bracket. The heating mechanism and the glass slide module are respectively installed on the incubation bracket. The heating mechanism includes a plurality of heating blocks arranged side by side. There is a row of glass slide positions on the glass slide module, and each heating block is respectively located directly below the corresponding glass slide position. The transverse movement mechanism is fixed on the incubation bracket. The addition part of the large reagent addition mechanism, the liquid inlet part of the cleaning mechanism and the waste suction part of the waste suction mechanism are respectively installed on the transverse movement mechanism. The transverse movement mechanism can drive the addition port of the large reagent addition mechanism, the liquid inlet port of the cleaning mechanism and the waste suction port of the waste suction mechanism to move along the straight line where the glass slide positions are distributed;

[0008] The cover slip conversion mechanism includes a two-axis moving mechanism, a cover slip support, a rotary conversion mechanism, a large reagent cover slip, and a micro reagent cover slip. The cover slip support is respectively provided with a row of large reagent cover slip mounting positions for mounting large reagent cover slips and a row of micro reagent cover slip mounting positions for mounting micro reagent cover slips. The straight line along which the large reagent cover slip mounting positions are distributed is parallel to the straight line along which the slide positions are distributed, and the straight line along which the micro reagent cover slip mounting positions are distributed is parallel to the straight line along which the slide positions are distributed. Each large reagent cover slip mounting position cooperates with the corresponding slide position, and each micro reagent cover slip mounting position cooperates with the corresponding slide position. The cover slip support is movably mounted on the two-axis moving mechanism. The cover slip support is located on one side of the slide module. The two-axis moving mechanism can drive the cover slip support to move along the vertical direction and the straight line perpendicular to the distribution of the slide positions respectively. The rotary conversion mechanism is fixed on the two-axis moving mechanism, and the rotary conversion mechanism drives the cover slip support to rotate;

[0009] The micro reagent sampling mechanism includes a three-axis moving mechanism and a sampling needle. The three-axis moving mechanism is mounted on the frame, and the sampling needle is mounted on the three-axis moving mechanism. The three-axis moving mechanism can drive the sampling needle to move along the straight line parallel to the distribution of the incubation units, along the straight line parallel to the distribution of the slide positions, and in the vertical direction respectively.

[0010] The heating mechanism includes a heating base and a lifting mechanism. A plurality of heating blocks are fixed side by side on the heating base. The fixed part of the lifting mechanism is fixed on the incubation support, and the moving part of the lifting mechanism is fixedly connected to the heating base. The lifting mechanism can drive the heating base to move up and down in the vertical direction.

[0011] A module fixing seat is provided at the top of the incubation support. The module fixing seat is of a frame structure. The module fixing seat includes two symmetrically arranged module support cross bars and two symmetrically arranged limiting rods. The two module support cross bars face each other directly and are parallel to each other. On one side of the two module support cross bars facing each other directly, module placement grooves are symmetrically provided. The two module placement grooves extend along the length directions of the two module support rods respectively. The cross section of the module placement groove is L-shaped. The slide module is square. The two sides in the length direction of the slide module are respectively clamped on the two module placement grooves. The two limiting rods face each other directly and are parallel to each other. The two limiting rods are respectively located directly below the two module support cross bars. The space formed by one limiting rod and the module support cross bar directly above it constitutes one of the upper and lower limiting spaces, and the space formed by the other limiting rod and the module support cross bar directly above it constitutes the other upper and lower limiting space. The heating base is square. The two side edges in the length direction of the heating base are respectively located in the two upper and lower limiting spaces. The lifting mechanism includes a pair of lifting cylinders. The cylinders of the pair of lifting cylinders are symmetrically fixed on the incubation support, and the piston rods of the pair of lifting cylinders are respectively fixedly connected to the heating base.

[0012] The described transverse movement mechanism includes a first Y-axis movement mechanism, a connecting vertical plate, and a positioning horizontal plate. The first Y-axis movement mechanism is installed on the incubation bracket. The bottom of the connecting vertical plate is connected to the moving part of the first Y-axis movement mechanism. One end of the positioning horizontal plate is connected to the top of the connecting vertical plate. The positioning horizontal plate is located above the glass slide module. The large-volume reagent dispensing mechanism includes a large-volume reagent container, a large-volume reagent dispensing peristaltic pump, a large-volume reagent dispensing hose, and a multi-channel dispenser. The large-volume reagent container is placed on the frame. The multi-channel dispenser includes multiple sample inlets and one sample outlet. The multi-channel dispenser is fixed on the positioning horizontal plate. The large-volume reagent dispensing hose is connected to the large-volume reagent peristaltic pump. The inlet of the large-volume reagent dispensing hose is connected to the large-volume reagent container. The outlet of the large-volume reagent dispensing hose is connected to one of the sample inlets. The cleaning mechanism includes a cleaning agent container, a cleaning agent adding hose, and a cleaning peristaltic pump. The cleaning agent container is placed on the frame. The cleaning agent adding hose is connected to the cleaning peristaltic pump. The inlet of the cleaning agent adding hose is connected to the cleaning agent container. The outlet of the cleaning agent adding hose is connected to another sample inlet. The waste suction mechanism includes a waste liquid container, a waste suction controller, a waste discharge hose, a waste discharge negative pressure pump, and a waste suction head. The waste liquid container is placed on the frame. The waste discharge hose is connected to the waste discharge negative pressure pump. The outlet of the waste discharge hose is connected to the waste liquid container. The inlet of the waste discharge hose is connected to the waste suction head. The fixed part of the waste suction controller is connected to the positioning horizontal plate. The moving part of the waste suction controller is connected to the waste suction head. The waste suction controller drives the waste suction head to move up and down in the vertical direction.

[0013] The described large-volume reagent dispensing mechanism further includes a preheater. The preheater is fixed on the positioning horizontal plate. The preheater is cylindrical. The large-volume reagent dispensing hose is wound around the outer wall of the preheater. The waste suction controller includes a push-pull electromagnet, a waste suction guide rod, a waste suction vertical support plate, and a waste suction head mounting plate. The bottom of the waste suction vertical support plate is fixed on the positioning horizontal plate. The lower end of the waste suction guide rod is fixed on the positioning horizontal plate. The push-pull electromagnet is fixed on the waste suction vertical support plate. The waste suction head mounting plate is L-shaped. The waste suction head is fixed on the horizontal part of the waste suction head mounting plate. The waste suction guide rod movably penetrates through the horizontal part of the waste suction head mounting plate. The push rod of the push-pull electromagnet is fixedly connected to the vertical part of the waste suction head mounting plate.

[0014] The described cover plate bracket includes a first cover plate mounting seat and a second cover plate mounting seat. The first cover plate mounting seat and the second cover plate mounting seat are strip-shaped. On one side in the length direction of the first cover plate mounting seat, there is a row of first cover plate mounting grooves for mounting a large number of reagent cover plates. There are a pair of positioning columns on the first cover plate mounting grooves, and a pair of positioning holes on the large number of reagent cover plates. Along the length direction of the first cover plate mounting seat, there is a row of magnets. Magnetically connected to the first cover plate mounting seat is a first cover plate that cooperates with the row of first cover plate mounting grooves. On one side in the length direction of the second cover plate mounting seat, there is a row of second cover plate mounting grooves for mounting micro reagent cover plates. There are a pair of positioning columns on the second cover plate mounting grooves, and a pair of positioning holes on the micro reagent cover plates. Along the length direction of the second cover plate mounting seat, there is a row of magnets. Magnetically connected to the second cover plate mounting seat is a second cover plate that cooperates with the row of second cover plate mounting grooves. The other side in the length direction of the first cover plate mounting seat is hinged to the other side in the length direction of the second cover plate mounting seat through a hinge. When the second cover plate mounting seat is in the horizontal position, the first cover plate mounting seat is located below the second cover plate mounting seat.

[0015] The described cover plate conversion mechanism further includes a cover plate cleaning tank, which is fixed on the side of the incubation bracket facing the first cover plate mounting seat. When the second cover plate mounting seat is in the horizontal position, the row of first cover plate mounting grooves on the first cover plate mounting seat is located directly above the cover plate cleaning tank. The rotation conversion mechanism includes a conversion motor. The two ends of the second cover plate mounting seat are respectively movably connected to two-axis moving mechanisms. The conversion motor is installed on the two-axis moving mechanisms and is connected to one end of the second cover plate mounting seat.

[0016] The described two-axis moving mechanism includes a first X-axis moving mechanism and a first Z-axis moving mechanism. The first X-axis moving mechanism is installed on the frame. The first X-axis moving mechanism is located on one side of the incubation bracket. The fixed part of the first Z-axis moving mechanism is installed on the moving part of the first X-axis moving mechanism. The two ends of the second cover plate mounting seat are respectively movably installed on the moving part of the first Z-axis moving mechanism through bearings. The conversion motor is installed on the moving part of the first Z-axis moving mechanism.

[0017] The described three-axis moving mechanism includes a second X-axis moving mechanism, a second Y-axis moving mechanism, and a second Z-axis moving mechanism. The second X-axis moving mechanism is installed on the frame. The fixed part of the second Y-axis moving mechanism is installed on the moving part of the second X-axis moving mechanism. The moving part of the second Z-axis moving mechanism is installed on the moving part of the second Y-axis moving mechanism. The micro reagent sampling mechanism further includes a micro reagent placement tank, which is installed on the frame. The sampling needle is fixed on the moving part of the second Z-axis moving mechanism, and the sampling needle is located above the micro reagent placement tank.

[0018] The described second Y-axis moving mechanism includes a housing, a second Y-axis motor, a driving pulley, a driven pulley, and a transmission belt. The housing is in the shape of a square strip. The bottom of one end of the housing is fixed on the moving part of the second X-axis moving mechanism. Strip-shaped slots are symmetrically provided at the top and bottom of the housing. The driving pulley and the driven pulley are respectively located inside the two ends of the housing, and the driven pulley is movably installed inside the housing. The second Y-axis motor is installed on one end of the housing close to the second X-axis moving mechanism. The driving pulley is sleeved on the output shaft of the second Y-axis motor. The driving pulley is connected to the driven pulley through the transmission belt. The second Z-axis moving mechanism includes a second Z-axis motor, a lifting rod, an engaging rod, and an adapter block. The second Z-axis motor is installed on one end of the housing close to the second X-axis moving mechanism. The cross-section of the engaging rod is in the shape of a regular polygon. The engaging rod is located inside the housing and is parallel to the moving direction of the transmission belt. One end of the engaging rod is connected to the output shaft of the second Z-axis motor, and the other end of the engaging rod is rotatably connected to the end of the housing far from the second X-axis moving mechanism. The adapter block is connected to the transmission belt. The lifting rod movably penetrates through the adapter block and respectively penetrates through the two slots. Tooth grooves are uniformly and densely arranged along the length direction of the lifting rod. The edges of the engaging rod are engaged with the tooth grooves on the lifting rod. The sampling needle is fixed to the lower end of the lifting rod.

[0019] Compared with the prior art, the beneficial effects and advantages of the present invention are as follows:

[0020] 1. This system can process dozens of samples simultaneously, enabling large-throughput batch staining, greatly improving the staining efficiency, and thus enhancing the detection efficiency of immunohistochemistry.

[0021] 2. This system adopts two sampling methods, namely micro reagent sampling and large reagent sampling, which can greatly save the reagents required in the staining process and reduce the staining cost.

[0022] 3. This device uses double cover glasses for antigen retrieval and staining. During the antigen retrieval process, a large reagent cover glass is used. The channel size on the large reagent cover glass is large and the capacity is large, which can accommodate sufficient antigen retrieval reagents, avoiding the problem of positive and negative slices caused by insufficient antigen amount in the antigen retrieval reagent. Moreover, exhaust holes communicating with the channels are provided on the large reagent cover glass, which can discharge air bubbles during the high-temperature heating process, avoiding the problem of flower slices caused by air bubble dry slices.

[0023] 4. Before adding the antigen retrieval reagent, this device uses a preheater to preheat the antigen retrieval reagent, which can precisely control the temperature during the antigen retrieval process and avoid the problem of positive and negative slices caused by the failure of the antigen retrieval reaction due to insufficient temperature.

[0024] 5. This device is provided with a waste suction mechanism, making each step of the reaction complete and improving the staining effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 It is a schematic structural diagram of a fully automatic immunohistochemical staining device.

[0026] Figure 2 It is an assembly drawing of a transverse movement mechanism, a large reagent sampling mechanism, a cleaning mechanism and a waste suction mechanism.

[0027] Figure 3 It is Figure 2 a partial schematic diagram of.

[0028] Figure 4 It is a schematic structural diagram of a coverslip conversion mechanism.

[0029] Figure 5 It is a schematic structural diagram of a coverslip support.

[0030] Figure 6 It is a schematic structural diagram of a coverslip support (with Figure 4 a different angle).

[0031] Figure 7 It is an assembly drawing of a heating mechanism, a glass slide module and an incubation support.

[0032] Figure 8 It is Figure 6 the front view of.

[0033] Figure 9 It is a schematic structural diagram of a three-axis movement mechanism.

[0034] Figure 10 It is a schematic internal structure diagram of a second Y-axis movement mechanism and a second Z-axis movement mechanism.

[0035] Figure 11 It is a schematic structural diagram of a large reagent coverslip.

[0036] Figure 12 It is a schematic structural diagram of a micro reagent coverslip.

[0037] Among them, 1 - frame, 2 - glass slide module, 3 - glass slide position, 4 - module support cross bar, 5 - limiting rod, 6 - module placement groove, 7 - heating base body, 8 - heating block, 9 - lifting cylinder, 10 - first Y-axis moving mechanism, 11 - connecting vertical plate, 12 - positioning horizontal plate, 13 - preheater, 14 - large reagent sampling hose, 15 - multi-channel sampler, 16 - cleaning agent adding hose, 17 - waste discharge hose, 18 - waste suction head, 19 - push-pull electromagnet, 20 - waste suction guide rod, 21 - waste suction vertical support plate, 22 - waste suction head mounting plate, 23 - first X-axis moving mechanism, 24 - first Z-axis moving mechanism, 25 - cover glass cleaning tank, 26 - first cover glass mounting seat, 27 - second cover glass mounting seat, 28 - first cover glass mounting groove, 29 - positioning column, 30 - magnet, 31 - first cover plate, 32 - hinge, 33 - incubation bracket, 34 - conversion motor, 35 - sampling needle, 36 - plunger pump, 37 - micro reagent sampling hose, 38 - micro reagent placement groove, 39 - second X-axis moving mechanism, 40 - outer housing, 41 - second Y-axis motor, 42 - driving pulley, 43 - driven pulley, 44 - transmission belt, 45 - second Z-axis motor, 46 - lifting rod, 47 - meshing rod, 48 - connecting block, 49 - tooth groove, 50 - slot, 51 - large reagent cover glass, 52 - micro reagent cover glass, 53 - first channel, 54 - large reagent adding hole, 55 - exhaust hole, 56 - positioning hole, 57 - second channel, 58 - micro reagent adding port, 59 - waste suction hole. Detailed implementation mode

[0038] The present invention will be described in detail below with reference to the accompanying drawings.

[0039] The structure of the full-automatic immunohistochemical staining device provided in this embodiment is as Figure 1 shown, including a frame 1, an incubation unit and a micro reagent sampling mechanism. The frame 1 provides support for the entire device, and the form can be various. There are 3 incubation units, and the 3 incubation units are sequentially fixed on the frame along a straight line.

[0040] The incubation unit includes a heating mechanism, a glass slide module 2, a large reagent sampling mechanism, a cleaning mechanism, a waste suction mechanism, a cover glass conversion mechanism, a transverse movement mechanism and an incubation bracket 33. The incubation bracket 33 provides support for the entire incubation unit.

[0041] As Figure 7 and Figure 8As shown in the figure, a module fixing base is provided at the top of the incubation bracket, and the module fixing base is of a frame structure. The module fixing base includes two symmetrically arranged module support crossbars 4 and two symmetrically arranged limiting rods 5. The two module support crossbars 4 are directly opposite and parallel to each other. On one side of the two directly opposite module support crossbars 4, module placement grooves 6 are symmetrically provided. The two module placement grooves 6 extend along the length directions of the two module support rods 4 respectively, and the cross-section of the module placement groove 6 is L-shaped. The slide module 2 is square, and a row of slide positions 3 is provided along its length direction. The straight line where the slide positions 3 are distributed is perpendicular to the straight line where the incubation units are distributed.

[0042] The slide module 2 is used to place the sample slides to be stained, and both sides of the slide module 2 in the length direction are respectively clamped on the two module placement grooves 6. The two limiting rods 5 are directly opposite and parallel to each other. The two limiting rods 5 are respectively located directly below the two module support crossbars 4. The space formed by one limiting rod 5 and the module support crossbar 4 directly above it constitutes one up-and-down limiting space, and the space formed by the other limiting rod 5 and the module support crossbar 4 directly above it constitutes another up-and-down limiting space.

[0043] The heating mechanism includes a heating base body 7 and a pair of lifting cylinders 9. The heating base body 7 is square, and a row of heating blocks 8 is provided along its length direction. The heating blocks 8 are square, and the shape and size of the heating blocks 8 match the shape and size of the slides. Each heating block 8 is respectively located directly below the corresponding slide position 3. The cylinders of a pair of lifting cylinders 9 are symmetrically fixed on the incubation bracket, and the piston rods of the pair of lifting cylinders 9 are respectively fixedly connected to the heating base body 7. The two side edges of the heating base body 7 in the length direction are respectively located in the two up-and-down limiting spaces.

[0044] As Figure 2 shown in the figure, the transverse movement mechanism includes a first Y-axis movement mechanism 10, a connecting vertical plate 11 and a positioning transverse plate 12. The first Y-axis movement mechanism 19 is installed on the incubation bracket. The bottom of the connecting vertical plate 11 is connected to the first Y-axis movement mechanism 19. One end of the positioning transverse plate 12 is connected to the top of the connecting vertical plate 11, and the positioning transverse plate 12 is located above the slide module 2. There are many ways of driving the first Y-axis movement mechanism 10. For example, the belt drive method can be used to drive the connecting vertical plate to move along the straight line where the slide positions are distributed.

[0045] As Figure 3As shown in the figure, the large-volume reagent sampling mechanism includes a large number of reagent containers, a large-volume reagent sampling peristaltic pump, a preheater 13, a large-volume reagent sampling hose 14, and a multi-channel sampler 15. The large number of reagent containers are placed on the rack. The preheater 13 is cylindrical and is fixed to the positioning cross plate 12. The large-volume reagent sampling hose 14 is wound around the outer wall of the preheater 13. The multi-channel sampler 13 includes a plurality of sampling ports and one sampling outlet, and the multi-channel sampler 13 is fixed to the positioning cross plate 12. The large-volume reagent sampling hose 14 is connected to the large-volume reagent peristaltic pump. The inlet of the large-volume reagent sampling hose 14 is connected to the large number of reagent containers, and the outlet of the large-volume reagent sampling hose 14 is connected to one of the sampling ports.

[0046] As Figure 3 shown in the figure, the cleaning mechanism includes a cleaning agent container, a cleaning agent filling hose 16, and a cleaning peristaltic pump. The cleaning agent container is placed on the rack. The cleaning agent filling hose 16 is connected to the cleaning peristaltic pump. The inlet of the cleaning agent filling hose 16 is connected to the cleaning agent container, and the outlet of the cleaning agent filling hose 16 is connected to another sampling port.

[0047] As Figure 3 shown in the figure, the waste suction mechanism includes a waste liquid container, a waste suction controller, a waste discharge hose 17, a waste discharge negative pressure pump, and a waste suction head 18. The waste liquid container is placed on the rack. The waste discharge hose 17 is connected to the waste discharge negative pressure pump. The waste suction controller includes a push-pull electromagnet 19, a waste suction guide rod 20, a waste suction vertical support plate 21, and a waste suction head mounting plate 22. The bottom of the waste suction vertical support plate 21 is fixed to the positioning cross plate 12. The lower end of the waste suction guide rod 20 is fixed to the positioning cross plate 12. The push-pull electromagnet 19 is fixed to the waste suction vertical support plate 21. The waste suction head mounting plate 22 is L-shaped. The waste suction head 18 is fixed to the horizontal part of the waste suction head mounting plate 22. The waste suction guide rod 20 movably penetrates through the horizontal part of the waste suction head mounting plate 22. The push rod of the push-pull electromagnet 19 is fixedly connected to the vertical part of the waste suction head mounting plate 22. The outlet of the waste discharge hose 17 is connected to the waste liquid container, and the inlet of the waste discharge hose 17 is connected to the waste suction head 18.

[0048] As Figure 4 shown in the figure, the cover sheet conversion mechanism includes a two-axis moving mechanism, a cover sheet cleaning tank 25, a cover sheet support, and a rotation conversion mechanism. As Figure 5 and Figure 6 shown in the figure, the cover sheet support includes a first cover sheet mounting seat 26 and a second cover sheet mounting seat 27. The first cover sheet mounting seat 26 and the second cover sheet mounting seat 27 are strip-shaped. On one side in the length direction of the first cover sheet mounting seat 26, there is a row of first cover sheet mounting grooves 28 for mounting a large number of reagent cover sheets. A pair of positioning columns 29 are provided on the first cover sheet mounting grooves 28. One end of the large number of reagent cover sheets is provided with a plastic connecting block, and positioning holes 56 that cooperate with the pair of positioning columns 29 are provided on the plastic connecting block. As Figure 11As shown, a square first channel 53 is formed on one surface of a large number of reagent coverslips 51. A large number of reagent filling holes 54 are formed on the large number of reagent coverslips 51, and the large number of reagent filling holes 54 communicate with the first channel 53. Three exhaust holes 55 are arranged in a straight line on the large number of reagent coverslips 51, and the three exhaust holes 55 respectively communicate with the first channel 53. A row of magnets 30 is arranged along the length direction of the first coverslip mounting seat 26, and a first cover plate 31 that cooperates with a row of first coverslip mounting grooves is magnetically connected to the first coverslip mounting seat 26.

[0049] On one side of the second coverslip mounting seat 27 in the length direction, there is a row of second coverslip mounting grooves for mounting micro reagent coverslips, and a pair of positioning posts are arranged on the second coverslip mounting grooves. One end of the micro reagent coverslip is provided with a plastic connecting block, and positioning holes 56 that cooperate with the pair of positioning posts are arranged on the plastic connecting block. As Figure 12 shown, a wedge-shaped second channel 57 is formed on one surface of the micro reagent coverslip 52, and the projection of the second channel 57 on the plane where one surface of the micro reagent coverslip 52 is located is square. The height of the second channel 57 decreases sequentially along the length direction of the micro reagent coverslip 52. The end with the lowest height of the second channel 57 is located on the edge of one width direction of the surface of the micro reagent coverslip 52, and the end with the lowest height of the second channel 57 is the micro reagent filling port 58. The micro reagent coverslip 52 is provided with waste suction holes, and the waste suction holes respectively communicate with the second channel 57. A row of magnets is arranged along the length direction of the second coverslip mounting seat 27, and a second cover plate that cooperates with a row of second coverslip mounting grooves is magnetically connected to the second coverslip mounting seat 27. Channels are respectively arranged on the large number of reagent coverslips and the micro reagent coverslips, and the depth of the channel on the large number of reagent coverslips is greater than the depth of the micro reagent coverslips.

[0050] The other side in the length direction of the first coverslip mounting seat 26 is hinged to the other side in the length direction of the second coverslip mounting seat 27 through a hinge 32. The first coverslip mounting seat 26 is provided with a row of counterweight blocks 33, and the coverslip cleaning tank 25 is fixed on the side of the incubation bracket facing the first coverslip mounting seat 26. When the second coverslip mounting seat 27 is in the horizontal position, the first coverslip mounting seat 26 is located below the second coverslip mounting seat 27, and a row of first coverslip mounting grooves on the first coverslip mounting seat 26 is located directly above the coverslip cleaning tank 25.

[0051] As Figure 4As shown in the figure, the two-axis moving mechanism includes a first X-axis moving mechanism 23 and a first Z-axis moving mechanism 24. The first X-axis moving mechanism 23 is installed on the frame. The first X-axis moving mechanism 23 is located on one side of the incubation bracket. The fixed part of the first Z-axis moving mechanism 24 is installed on the moving part of the first X-axis moving mechanism 23. Both ends of the second cover plate mounting seat 27 are movably installed on the moving part of the first Z-axis moving mechanism 24 through bearings. There are many ways to drive the first X-axis moving mechanism 23 and the first Z-axis moving mechanism 24, and a way similar to the belt drive of the second Y-axis moving mechanism can be adopted. The first X-axis moving mechanism 23 drives the cover plate bracket to move along a straight line parallel to the line perpendicular to the slide position 3, and the first Z-axis moving mechanism 24 drives the cover plate bracket to move along the vertical direction.

[0052] The rotation conversion mechanism includes a conversion motor 34. The conversion motor 34 is installed on the moving part of the first Z-axis moving mechanism 24, and the conversion motor 34 is connected to one end of the second cover plate mounting seat 27.

[0053] As Figure 9 shown, the micro reagent sampling mechanism includes a three-axis moving mechanism, a sampling needle 35, a plunger pump 36, a micro reagent sampling hose 37, and a micro reagent placement groove 38. The micro reagent placement groove 38 is installed on the frame 1. One end of the micro reagent sampling hose 37 is connected to the plunger pump 36, and the other end is connected to the sampling needle 35. The three-axis moving mechanism includes a second X-axis moving mechanism 39, a second Y-axis moving mechanism, and a second Z-axis moving mechanism. The second X-axis moving mechanism 39 is installed on the frame. There are many ways to drive the second X-axis moving mechanism 39, and a way similar to the belt drive of the second Y-axis moving mechanism can be adopted. The second X-axis moving mechanism can drive the second Y-axis moving mechanism to move along a straight line where the incubation unit is distributed.

[0054] As Figure 10 shown, the second Y-axis moving mechanism includes a housing 40, a second Y-axis motor 41, a driving pulley 42, a driven pulley 43, and a transmission belt 44. The housing 40 is in the shape of a square strip. The bottom of one end of the housing 40 is fixed to the moving part of the second X-axis moving mechanism 39. Strip-shaped slot grooves 50 are symmetrically arranged at the top and bottom of the housing 40. The driving pulley 42 and the driven pulley 43 are respectively located inside both ends of the housing 40, and the driven pulley 43 is movably installed inside the housing 40. The second Y-axis motor 41 is installed on one end of the housing close to the second X-axis moving mechanism 39. The driving pulley 42 is sleeved on the output shaft of the second Y-axis motor 41, and the driving pulley 42 is connected to the driven pulley 43 through the transmission belt 44. The moving direction of the transmission belt 44 is perpendicular to the straight line where the incubation unit is distributed.

[0055] The second Z-axis moving mechanism includes a second Z-axis motor 45, a lifting rod 46, an engaging rod 47 and an adapter block 48. The second Z-axis motor 45 is installed at one end of the outer housing 40 close to the second X-axis moving mechanism 39. The cross-section of the engaging rod 47 is in the shape of a regular polygon. The engaging rod 47 is located inside the outer housing 40 and is parallel to the moving direction of the conveyor belt 44. One end of the engaging rod 47 is connected to the output shaft of the second Z-axis motor 45, and the other end of the engaging rod 47 is rotatably connected to one end of the outer housing 40 away from the second X-axis moving mechanism 39. The adapter block 48 is connected to the conveyor belt 44. The lifting rod 46 movably passes through the adapter block 48. Tooth grooves 49 are uniformly and densely arranged along the length direction of the lifting rod 46. The edges of the engaging rod 47 are engaged with the tooth grooves 49 on the lifting rod 46. The sampling needle 35 is fixed to the lower end of the lifting rod. The output shaft of the second Z-axis motor 45 rotates to drive the engaging rod 47 to rotate. The rotation of the engaging rod 47 drives the lifting rod 46 to move up and down. The lifting rod 46 drives the sampling needle 35 to move up and down for sampling.

[0056] The detection method of the above-mentioned fully automatic immunohistochemical staining device is as follows:

[0057] 1. Add a large amount of reagent into a large reagent container, add deionized water into the cleaning agent container, place the micro reagent bottles containing micro reagent one, micro reagent two, micro reagent three, micro reagent four, micro reagent five and micro reagent six in sequence in the micro reagent placement groove, then initialize the whole device, fill each liquid path, and make each movable mechanism return to the initial position;

[0058] 2. For the convenience of description, take one incubation unit as an example for explanation. Place the sample slide used in the experiment on the slide module, then place the slide module 2 on the module fixing seat, turn on the lifting air cylinder 9. The lifting air cylinder 9 drives the heating seat body 7 to move downward to a set distance, and then the lifting air cylinder 9 is turned off. At this time, each heating block 8 on the heating seat body is respectively located directly below the corresponding sample slide, and each heating block 8 is in contact with the corresponding sample slide. Install the large reagent coverslips required for the experiment on the corresponding first coverslip mounting grooves 28 respectively, and magnetically connect the first cover plate 31 to the first coverslip mounting seat 26 to fix each large reagent coverslip. Install the micro reagent coverslips required for the experiment on the corresponding second coverslip mounting grooves respectively, and magnetically connect the second coverslip to the second coverslip mounting seat to fix each micro reagent coverslip;

[0059] 3. The heating mechanism starts to work, heat the temperature of each heating block 8 to 68 °C for preheating the slide. After the slide preheating is completed, the heating mechanism stops working, and the preheater 13 starts to work, and the temperature rises to 85 °C;

[0060] 4. The conversion motor 34 starts to operate, driving the first cover plate mounting seat 26 to rotate. When the first cover plate mounting seat 26 is in the horizontal position, the conversion motor 34 stops operating. The first X-axis moving mechanism 23 starts to operate, moving the first cover plate mounting seat 26 directly above the glass slide module 2. The first Z-axis moving mechanism 24 starts to operate, moving the first cover plate mounting seat 26 downward. When each large reagent cover plate is respectively covered on the corresponding sample glass slide, the first Z-axis moving mechanism 24 stops operating;

[0061] 5. The first Y-axis moving mechanism 10 starts to operate, driving the positioning cross plate 12 to move along the straight line where the sample glass slides are distributed. When the positioning cross plate 12 moves to the set position, the first Y-axis moving mechanism 10 stops operating. The large reagent peristaltic pump starts to operate, adding a large amount of reagent to the corresponding sample glass slides through the sample outlet of the multi-channel sampler 15. This process repeats until all sample glass slides are successfully sampled. The first Y-axis moving mechanism 10 returns to the initial position, and the large reagent peristaltic pump stops operating. The heating mechanism starts to operate. After heating and incubating for 40 minutes, both the preheater 13 and the heating mechanism stop operating, and the temperature is decreased for 20 minutes until the temperature reduction ends;

[0062] 6. The first Z-axis moving mechanism 24 starts to operate, moving the first cover plate mounting seat 26 upward. When the first cover plate mounting seat 26 moves a set distance, the first Z-axis moving mechanism 24 stops operating. The first X-axis moving mechanism 23 starts to operate, moving the first cover plate mounting seat 26 to the initial position;

[0063] 7. The conversion motor 34 starts to operate, driving the second cover plate mounting seat 27 to rotate. When the second cover plate mounting seat 27 is in the horizontal position, the conversion motor 34 stops operating. The first X-axis moving mechanism 23 starts to operate, moving the first cover plate mounting seat 26 directly above the glass slide module 2. The first Z-axis moving mechanism 24 starts to operate, driving the first cover plate mounting seat 26 downward to the set position. When each micro-reagent cover plate is respectively covered on the corresponding sample glass slide, the first Z-axis moving mechanism 24 stops operating; At this time, the large reagent cover plates on the first cover plate mounting seat 26 are being cleaned in the cover plate cleaning tank 25;

[0064] 8. The first Y-axis moving mechanism 10 starts to work. The first Y-axis moving mechanism 10 drives the positioning cross plate 12 to move along the straight line where the sample slides are distributed. When the positioning cross plate 12 moves to the set position, the first Y-axis moving mechanism 10 stops working. The push-pull electromagnet 19 starts to work. The push-pull electromagnet 19 pushes the waste suction head 18 to move downward to the set position. The push-pull electromagnet 19 stops working. The waste discharge negative pressure pump starts to work to extract the waste liquid on the corresponding sample slide. After the waste discharge negative pressure pump works for the set time, it stops working. The push-pull electromagnet 19 is turned off. The push-pull electromagnet 19 drives the waste suction head 18 to move upward to the set position. The peristaltic pump is turned on, and deionized water is added to the corresponding sample slide through the sample outlet of the multi-channel sampler 15 for cleaning. Then the push-pull electromagnet 19 starts to work for waste suction, and the above waste suction and cleaning actions are repeated several times until a large amount of residual reagent on the corresponding sample slide is cleaned;

[0065] 9. Repeat step 8 until all sample slides are cleaned, and the first Y-axis moving mechanism 10 returns to the initial position;

[0066] 10. The second Y-axis motor 41 starts to work. The second Y-axis motor 41 drives the sampling needle 35 to move above the micro reagent bottle containing the first micro reagent. The second Y-axis motor 41 stops working. The second Z-axis motor 45 starts to work. The second Z-axis motor 45 drives the sampling needle 35 to move downward below the liquid level of the micro reagent bottle containing the first micro reagent. The second Z-axis motor 45 stops working. The plunger pump 36 starts to work to extract the first micro reagent to the set volume. The second Z-axis motor starts to work. The second Z-axis motor 45 drives the sampling needle 35 to move upward to the set position. The second Z-axis motor 45 stops working. The second X-axis moving mechanism 39 starts to work to drive the sampling needle 35 to move a set distance. At this time, the sampling needle 35 is above the slide module 2. Then the second Y-axis motor 41 and the second Z-axis motor 45 perform actions in sequence, and the first micro reagent is added to the corresponding sample slide through the corresponding micro reagent cover slip inlet. After incubation for a period of time, waste suction and cleaning actions are performed to clean the residual first micro reagent on the sample slide;

[0067] 11. Repeat step 10, and add the second micro reagent, the third micro reagent, the fourth micro reagent, the fifth micro reagent, and the sixth micro reagent to each sample slide in sequence for incubation;

[0068] 12. The first Z-axis moving mechanism 24 and the first X-axis moving mechanism 23 perform actions in sequence, and the cover slip support returns to the initial position. At this time, the entire staining step is completed.

Claims

1. A fully automatic immunohistochemical staining device, characterized in that: it includes a frame, an incubation unit and a micro reagent injection mechanism. There are multiple incubation units, and the multiple incubation units are sequentially fixed on the frame along a straight line; The incubation unit includes a heating mechanism, a slide module, a large reagent adding mechanism, a cleaning mechanism, a waste suction mechanism, a coverslip conversion mechanism, a transverse movement mechanism and an incubation bracket. The heating mechanism and the slide module are respectively installed on the incubation bracket. The heating mechanism includes multiple heating blocks arranged side by side. There is a row of slide positions on the slide module, and each heating block is respectively located directly below the corresponding slide position. The transverse movement mechanism is fixed on the incubation bracket. The adding part of the large reagent adding mechanism, the liquid inlet part of the cleaning mechanism and the waste suction part of the waste suction mechanism are respectively installed on the transverse movement mechanism. The transverse movement mechanism can drive the adding port of the large reagent adding mechanism, the liquid inlet port of the cleaning mechanism and the waste suction port of the waste suction mechanism to move along the straight line where the slide positions are distributed; The coverslip conversion mechanism includes a two-axis movement mechanism, a coverslip bracket, a rotation conversion mechanism, large reagent coverslips and micro reagent coverslips. The coverslip bracket is respectively provided with a row of large reagent coverslip installation positions for installing large reagent coverslips and a row of micro reagent coverslip installation positions for installing micro reagent coverslips. The straight line where the large reagent coverslip installation positions are distributed is parallel to the straight line where the slide positions are distributed, and the straight line where the micro reagent coverslip installation positions are distributed is parallel to the straight line where the slide positions are distributed. Each large reagent coverslip installation position cooperates with the corresponding slide position, and each micro reagent coverslip installation position cooperates with the corresponding slide position. The coverslip bracket is movably installed on the two-axis movement mechanism. The coverslip bracket is located on one side of the slide module. The two-axis movement mechanism can drive the coverslip bracket to move respectively along the vertical direction and the straight line perpendicular to the distribution of the slide positions. The rotation conversion mechanism is fixed on the two-axis movement mechanism, and the rotation conversion mechanism drives the coverslip bracket to rotate; The micro reagent injection mechanism includes a three-axis movement mechanism and an injection needle. The three-axis movement mechanism is installed on the frame, and the injection needle is installed on the three-axis movement mechanism. The three-axis movement mechanism drives the injection needle to move respectively along the straight line parallel to the distribution of the incubation units, along the straight line parallel to the distribution of the slide positions and in the vertical direction.

2. The fully automatic immunohistochemical staining device according to claim 1, characterized in that: the heating mechanism includes a heating base and a lifting mechanism. Multiple heating blocks are fixed side by side on the heating base. The fixed part of the lifting mechanism is fixed on the incubation bracket, and the moving part of the lifting mechanism is fixedly connected with the heating base. The lifting mechanism can drive the heating base to move up and down in the vertical direction.

3. The fully automatic immunohistochemical staining device according to claim 2, characterized in that: The top of the incubation bracket is provided with a module fixing seat, and the module fixing seat is of a frame structure. The module fixing seat includes two symmetrically arranged module support cross bars and two symmetrically arranged limiting rods. The two module support cross bars face each other directly and are parallel to each other. On one side of the two module support cross bars facing each other directly, module placement grooves are symmetrically provided. The two module placement grooves extend along the length directions of the two module support bars respectively. The cross section of the module placement groove is L-shaped. The glass slide module is square. The two sides of the glass slide module in the length direction are respectively clamped on the two module placement grooves. The two limiting rods face each other directly and are parallel to each other. The two limiting rods are respectively located directly below the two module support cross bars. The space formed by one limiting rod and the module support cross bar directly above it constitutes one up-and-down limiting space, and the space formed by the other limiting rod and the module support cross bar directly above it constitutes another up-and-down limiting space. The heating seat body is square. The two side edges of the heating seat body in the length direction are respectively located in the two up-and-down limiting spaces. The lifting mechanism includes a pair of lifting cylinders. The cylinders of the pair of lifting cylinders are symmetrically fixed on the incubation bracket, and the piston rods of the pair of lifting cylinders are respectively fixedly connected to the heating seat body.

4. The fully automatic immunohistochemical staining device according to claim 1, characterized in that: The transverse movement mechanism includes a first Y-axis movement mechanism, a connecting vertical plate and a positioning transverse plate. The first Y-axis movement mechanism is installed on the incubation bracket. The bottom of the connecting vertical plate is connected to the moving part of the first Y-axis movement mechanism. One end of the positioning transverse plate is connected to the top of the connecting vertical plate. The positioning transverse plate is located above the glass slide module. The large amount of reagent adding mechanism includes a large amount of reagent container, a large amount of reagent adding peristaltic pump, a large amount of reagent adding flexible hose and a multi-channel sampler. The large amount of reagent container is placed on the frame. The multi-channel sampler includes a plurality of sample inlets and one sample outlet. The multi-channel sampler is fixed on the positioning transverse plate. The large amount of reagent adding flexible hose is connected to the large amount of reagent peristaltic pump. The inlet of the large amount of reagent adding flexible hose is connected to the large amount of reagent container, and the outlet of the large amount of reagent adding flexible hose is connected to one of the sample inlets. The cleaning mechanism includes a cleaning agent container, a cleaning agent adding liquid flexible hose and a cleaning peristaltic pump. The cleaning agent container is placed on the frame. The cleaning agent adding liquid flexible hose is connected to the cleaning peristaltic pump. The inlet of the cleaning agent adding liquid flexible hose is connected to the cleaning agent container, and the outlet of the cleaning agent adding liquid flexible hose is connected to another sample inlet. The waste suction mechanism includes a waste liquid container, a waste suction controller, a waste discharge hose, a waste discharge negative pressure pump and a waste suction head. The waste liquid container is placed on the frame. The waste discharge hose is connected to the waste discharge negative pressure pump. The outlet of the waste discharge hose is connected to the waste liquid container. The inlet of the waste discharge hose is connected to the waste suction head. The fixed part of the waste suction controller is connected to the positioning transverse plate, and the moving part of the waste suction controller is connected to the waste suction head. The waste suction controller drives the waste suction head to move up and down in the vertical direction.

5. The fully automatic immunohistochemical staining device according to claim 4, characterized in that: The large reagent loading mechanism further includes a preheater which is fixed on the positioning cross plate. The preheater is cylindrical, and the large reagent loading hose is wound around the outer wall of the preheater. The waste suction controller includes a push-pull electromagnet, a waste suction guide rod, a waste suction vertical support plate and a waste suction head mounting plate. The bottom of the waste suction vertical support plate is fixed on the positioning cross plate, the lower end of the waste suction guide rod is fixed on the positioning cross plate, the push-pull electromagnet is fixed on the waste suction vertical support plate, the waste suction head mounting plate is L-shaped, the waste suction head is fixed on the horizontal part of the waste suction head mounting plate, the waste suction guide rod movably penetrates through the horizontal part of the waste suction head mounting plate, and the push rod of the push-pull electromagnet is fixedly connected with the vertical part of the waste suction head mounting plate.

6. The full-automatic immunohistochemical staining device according to claim 1, characterized in that: The cover slip bracket includes a first cover slip mounting seat and a second cover slip mounting seat. The first cover slip mounting seat and the second cover slip mounting seat are strip-shaped. On one side in the length direction of the first cover slip mounting seat, there is a row of first cover slip mounting grooves for mounting large reagent cover slips. There are a pair of positioning posts on the first cover slip mounting grooves, and a pair of positioning holes on the large reagent cover slips. Along the length direction of the first cover slip mounting seat, there is a row of magnets. A first cover plate that cooperates with the row of first cover slip mounting grooves is magnetically connected to the first cover slip mounting seat. On one side in the length direction of the second cover slip mounting seat, there is a row of second cover slip mounting grooves for mounting micro reagent cover slips. There are a pair of positioning posts on the second cover slip mounting grooves, and a pair of positioning holes on the micro reagent cover slips. Along the length direction of the second cover slip mounting seat, there is a row of magnets. A second cover plate that cooperates with the row of second cover slip mounting grooves is magnetically connected to the second cover slip mounting seat. The other side in the length direction of the first cover slip mounting seat and the other side in the length direction of the second cover slip mounting seat are hinged by a hinge. When the second cover slip mounting seat is in the horizontal position, the first cover slip mounting seat is located below the second cover slip mounting seat.

7. The full-automatic immunohistochemical staining device according to claim 1, characterized in that: The cover slip conversion mechanism further includes a cover slip cleaning tank which is fixed on the side of the incubation bracket facing the first cover slip mounting seat. When the second cover slip mounting seat is in the horizontal position, the row of first cover slip mounting grooves on the first cover slip mounting seat is located directly above the cover slip cleaning tank. The rotation conversion mechanism includes a conversion motor. The two ends of the second cover slip mounting seat are respectively movably connected to the two-axis movement mechanism. The conversion motor is installed on the two-axis movement mechanism and is connected to one end of the second cover slip mounting seat.

8. The full-automatic immunohistochemical staining device according to claim 7, characterized in that: The two-axis movement mechanism includes a first X-axis movement mechanism and a first Z-axis movement mechanism. The first X-axis movement mechanism is installed on the frame and is located on one side of the incubation bracket. The fixed part of the first Z-axis movement mechanism is installed on the moving part of the first X-axis movement mechanism. The two ends of the second cover slip mounting seat are respectively movably installed on the moving part of the first Z-axis movement mechanism through bearings. The conversion motor is installed on the moving part of the first Z-axis movement mechanism.

9. The full-automatic immunohistochemical staining device according to claim 1, It is characterized in that: The three-axis moving mechanism includes a second X-axis moving mechanism, a second Y-axis moving mechanism and a second Z-axis moving mechanism. The second X-axis moving mechanism is installed on the frame. The fixed part of the second Y-axis moving mechanism is installed on the moving part of the second X-axis moving mechanism. The moving part of the second Z-axis moving mechanism is installed on the moving part of the second Y-axis moving mechanism. The micro reagent sampling mechanism further includes a micro reagent placement groove which is installed on the frame. The sampling needle is fixed on the moving part of the second Z-axis moving mechanism, and the sampling needle is located above the micro reagent placement groove.

10. The fully automatic immunohistochemical staining device according to claim 9, It is characterized in that: The second Y-axis moving mechanism includes an outer housing, a second Y-axis motor, a driving pulley, a driven pulley and a transmission belt. The outer housing is in a square strip shape. The bottom of one end of the outer housing is fixed on the moving part of the second X-axis moving mechanism. Strip-shaped slot grooves are symmetrically arranged at the top and bottom of the outer housing. The driving pulley and the driven pulley are respectively located inside the two ends of the outer housing, and the driven pulley is movably installed inside the outer housing. The second Y-axis motor is installed on one end of the outer housing close to the second X-axis moving mechanism. The driving pulley is sleeved on the output shaft of the second Y-axis motor. The driving pulley is connected to the driven pulley through the transmission belt. The second Z-axis moving mechanism includes a second Z-axis motor, a lifting rod, an engaging rod and a connecting block. The second Z-axis motor is installed on one end of the outer housing close to the second X-axis moving mechanism. The cross section of the engaging rod is in a regular polygon shape. The engaging rod is located inside the outer housing and is parallel to the moving direction of the transmission belt. One end of the engaging rod is connected to the output shaft of the second Z-axis motor, and the other end of the engaging rod is rotatably connected to one end of the outer housing away from the second X-axis moving mechanism. The connecting block is connected to the transmission belt. The lifting rod movably penetrates through the connecting block and respectively penetrates through the two slot grooves. Tooth grooves are uniformly and densely arranged along the length direction of the lifting rod. The edge of the engaging rod meshes with the tooth grooves on the lifting rod. The sampling needle is fixed on the lower end of the lifting rod.

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