Liquid path system for full-automatic multi-piece independent FISH dyeing equipment and full-automatic multi-piece independent FISH dyeing equipment

Abstract

The invention belongs to the technical field related to medical in-vitro diagnosis molecular pathology instruments, and discloses a liquid path system for full-automatic multi-piece independent FISH staining equipment and the full-automatic multi-piece independent FISH staining equipment. The liquid path system comprises a first reagent liquid path system, a second reagent liquid path system, a third reagent liquid path system and an independent sample reaction bin; the independent sample reaction bin comprises a plurality of independent reaction tanks and a sample clamping bin, and the plurality of reaction tanks are arranged in parallel; one sample clamping bin is arranged in each reaction tank, the sample clamping bins are used for clamping sample slides, and the first reagent liquid path system, the second reagent liquid path system and the third reagent liquid path system are used for independently adding reagents into the sample clamping bins. According to the invention, cross contamination during dyeing of multiple samples is avoided.

Description

Technical Field

[0001] The present invention belongs to the technical field related to medical in vitro diagnostic molecular pathology instruments, and more specifically, relates to a liquid path system for a fully automatic multi-chip independent FISH staining device and a fully automatic multi-chip independent FISH staining device. Background Art

[0002] Fluorescence in situ hybridization (FISH) is an effective method for detecting nucleic acid sequences at the molecular level. It is widely used in the diagnosis, treatment and research of malignant tumors such as breast cancer, lymphoma and bladder cancer. The key steps of FISH experiments include baking, dewaxing, washing, permeabilization, water treatment, enzyme digestion, dehydration, denaturation, hybridization, elution, counterstaining and sealing. This process requires a variety of experimental instruments and reagents. Manual experimental operations are cumbersome and prone to errors, resulting in poor staining consistency. Secondly, depending on the probe reagents used or the detection requirements, staining usually takes more than 10 to 16 hours. Manual FISH experiments will consume a lot of manpower and material resources, and the detection efficiency is greatly limited.

[0003] In addition, the current staining equipment in the field basically adopts vat dyeing, that is, multiple samples are stained in the same staining vat, which is easy to cause biological contamination between samples. At present, it is urgent to develop fully automatic multi-chip independent FISH staining equipment to replace manual work and realize the process standardization and precision of the staining process. The liquid circuit system is an important part of the fully automatic FISH staining equipment. It is crucial to realize the independent staining of multiple processes and multiple samples. It is urgent to carry out its design and implementation to realize the full automation of the FISH staining process. Summary of the invention

[0004] In view of the above defects or improvement needs of the prior art, the present invention provides a liquid path system for a fully automatic multi-chip independent FISH staining device and a fully automatic multi-chip independent FISH staining device, which aims to solve the problem of cross contamination when staining multiple samples.

[0005] To achieve the above-mentioned purpose, according to one aspect of the present invention, a liquid circuit system for a fully automatic multi-slide independent FISH staining device is provided, wherein the liquid circuit system includes a first reagent liquid circuit system, a second reagent liquid circuit system, a third reagent liquid circuit system and an independent sample reaction chamber, wherein the independent sample reaction chamber includes a plurality of independent reaction slots and a sample clamping chamber, and the plurality of reaction slots are arranged in parallel; one sample clamping chamber is arranged in each reaction slot, and the sample clamping chamber is used to clamp sample slides, and the first reagent liquid circuit system, the second reagent liquid circuit system and the third reagent liquid circuit system are used to independently add reagents to the sample clamping chamber.

[0006] Furthermore, the first reagent liquid path system includes at least one first reagent supply unit, the first reagent supply unit includes a first reagent bottle, a peristaltic pump and a flow meter, the peristaltic pump is connected to the first reagent bottle and the flow meter, the flow meter is connected to the first reagent discharge port through a pipeline, and the first reagent discharge port is used to introduce the reagent from the first reagent supply unit into the corresponding reaction tank.

[0007] Furthermore, at least six first reagent supply units are provided; and the volume of the first reagent bottle is 300 mL to 1000 mL.

[0008] Furthermore, the independent sample reaction chamber is provided with at least 10 reaction slots, and the reaction slots are arranged in parallel.

[0009] Furthermore, the second reagent liquid path system includes at least one second reagent supply unit, which includes a second reagent bottle, a pipette tip, a microinjection pump and a plurality of disposable pipette tips arranged on the equipment reaction platform, the tail of the pipette tip is connected to the microinjection pump, the pipette tip is used to suck up the disposable pipette tip and insert the disposable pipette tip into the second reagent bottle, and the reagent in the second reagent bottle is pumped into the corresponding disposable pipette tip through the microinjection pump; the second reagent liquid path system pumps out the second reagent by removing the disposable pipette tip from the second reagent bottle and moving it into the reaction tank.

[0010] Furthermore, at least 10 second reagent supply units are provided; and the volume of the second reagent bottle is 0.5 mL to 10 mL.

[0011] Furthermore, the third reagent liquid path system includes at least one third reagent supply unit, the third reagent supply unit includes a third reagent air pump, a third reagent bottle, a third reagent ventilation tube, a third reagent infusion tube and a third reagent nozzle, the third reagent air pump is connected to the third reagent bottle through the third reagent ventilation tube, and the third reagent bottle is connected to the third reagent nozzle through the third reagent infusion tube.

[0012] Furthermore, the volume of the third reagent bottle is 20 mL to 200 mL; and the reaction tanks are centrally arranged on the equipment reaction platform.

[0013] Furthermore, the emptying of the reagent in the reaction tank is completed by a reaction tank reagent emptying module; the reaction tank reagent emptying module includes a strip air nozzle, a reagent emptying module air pump and a reagent emptying module air pipe, and the reagent emptying module air pipe connects the reagent emptying module air pump and the strip air nozzle; the reaction tank emptying module emits a strip airflow through the strip air nozzle to flush the residual reagent in the reaction tank.

[0014] The present invention also provides a fully automatic multi-chip independent FISH staining device, comprising the liquid path system and controller for the fully automatic multi-chip independent FISH staining device as described above, wherein the controller is connected to the liquid path system.

[0015] In general, compared with the prior art, the above technical solutions conceived by the present invention mainly have the following beneficial effects in the liquid path system for a fully automatic multi-chip independent FISH staining device and the fully automatic multi-chip independent FISH staining device provided by the present invention:

[0016] 1. The liquid path system for the fully automatic multi-slice independent FISH staining device provided by the present invention can fully automatically complete the process-based dripping and discharge of independent reagents for multiple samples, avoiding the cross-contamination problem of staining multiple samples in the same staining tank and improving the reliability of the experimental results.

[0017] 2. The liquid path system for the fully automatic FISH staining equipment of the present invention can fully automatically inject or discharge reagents into or out of the reaction tank, avoiding the disadvantages of different operating levels of personnel, lack of standardized operations, and large experimental randomness, thereby reducing experimental differences, improving experimental efficiency, and making the detection operation meet the traceable GLP experimental requirements.

[0018] 3. The present invention is provided with an independent sample reaction chamber, which includes a plurality of reaction tanks arranged in parallel. When completing the staining process of multiple samples, each sample is independently processed and liquid is added. The reaction environment of each sample is isolated, avoiding the risk of contamination caused by staining using a staining jar.

[0019] 4. In view of the differences in reagent types, dosages, and precision requirements for each step in the fully automated FISH process, the first reagent liquid circuit system, the second reagent liquid circuit system, and the third reagent liquid circuit system are set up for different dosages, reagent types, and precision requirements; among them, the control accuracy of the first reagent liquid circuit system can reach ±0.5mL, the control accuracy of the second reagent liquid circuit system can reach ±1μL, and the control accuracy of the third reagent liquid circuit system can reach ±0.2mL; the first reagent liquid circuit system is for reagents with high consumption and low prices, the second reagent liquid circuit system is for reagents with low consumption and high prices, and the third reagent liquid circuit system is for reagents with moderate consumption and prices. It can achieve precise control of the dosage of various reagents, save reagent consumption, and thus reduce the cost of FISH experiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 It is a schematic diagram of a liquid path system for a fully automatic multi-chip independent FISH staining device provided by the present invention;

[0021] Figure 2 yes Figure 1A schematic diagram of a reaction tank reagent emptying module of a liquid path system for a fully automatic multi-chip independent FISH staining device;

[0022] Figure 3 yes Figure 1 Schematic diagram of the third reagent liquid path system of the liquid path system for a fully automatic multi-slice independent FISH staining device.

[0023] In all the drawings, the same figure numbers are used to represent the same elements or structures, wherein: 1-first reagent bottle, 2-peristaltic pump, 3-flow meter, 4-equipment reaction platform, 5-reaction tank, 6-waste diversion tube, 7-microinjection pump, 8-first reagent discharge port, 9-pipette tip, 10-disposable pipette tip, 11-well plate, 12-second reagent bottle, 21-strip air nozzle, 22-reagent emptying module air pump, 23-reagent emptying module air pipe, 31-third reagent bottle, 32-third reagent nozzle, 33-third reagent infusion tube, 34-third reagent ventilation tube, 35-third reagent air pump. DETAILED DESCRIPTION

[0024] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention. In addition, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0025] The present invention provides a liquid circuit system for a fully automatic multi-chip independent FISH staining device, which can fully automatically complete the process-based dripping and discharge of reagents for multiple samples, avoiding the cross-contamination problem of multiple samples dyeing in the same staining tank, and improving the reliability of experimental results; at the same time, it avoids the disadvantages of different operating levels of personnel, lack of standardized operations, and large experimental randomness, thereby reducing experimental differences, improving experimental efficiency, and making the detection operation meet the traceable GLP experimental requirements. In addition, the liquid circuit system can also realize the precise control of the amount of various reagents, save reagent consumption, and reduce the cost of FISH experiments.

[0026] See also Figure 1 , Figure 2 and Figure 3The liquid circuit system includes a first reagent liquid circuit system, a second reagent liquid circuit system, a third reagent liquid circuit system and an independent sample reaction chamber. The independent sample reaction chamber includes a plurality of independent reaction slots 5 and a sample clamping chamber. The plurality of reaction slots 5 are arranged in parallel. One sample clamping chamber is arranged in each reaction slot 5. The sample clamping chamber is used to clamp sample slides. The first reagent liquid circuit system, the second reagent liquid circuit system and the third reagent liquid circuit system are used to independently add reagents to the sample clamping chamber.

[0027] The first reagent liquid path system includes at least one first reagent supply unit, which includes a first reagent bottle 1, a peristaltic pump 2 and a flow meter 3. The peristaltic pump 2 is connected to the first reagent bottle 1 and the flow meter 3. The flow meter 3 is connected to a first reagent discharge port 8 through a pipeline. The first reagent discharge port 8 is used to introduce the reagent from the first reagent supply unit into the corresponding reaction tank 5, and drain the reagent after the reaction is completed.

[0028] In one embodiment, at least six first reagent supply units are provided, and six first reagent bottles 1 are connected to six peristaltic pumps 2 and six flow meters 3 through pipelines, and the liquid outlets of the six flow meters 3 are connected to four drainage ports. The liquid outlets of the first flow meter 3 and the second flow meter 3 are combined and connected to the first drainage port, the liquid outlets of the third flow meter 3 and the fourth flow meter 3 are combined and connected to the second drainage port, the liquid outlet of the fifth flow meter 3 is connected to the third drainage port, and the liquid outlet of the sixth flow meter 3 is connected to the fourth drainage port. The independent sample reaction chamber is provided with at least 10 reaction tanks 5, and the reaction tanks 5 are arranged in parallel; each reaction tank 5 can accommodate a sample slice and complete the independent reaction processes such as immersion and heating required for the staining experiment. A waste liquid diversion pipe 6 is provided below the reaction platform 4 of the equipment, and the waste liquid diversion pipe 6 is connected to the waste liquid barrel through a waste liquid pipe.

[0029] Different reagents enter the reaction tank 5 at different time periods, and after heating, cooling, flowing, incubating, soaking, and washing the sample, a fully automatic sample experiment process is completed. The volume of the first reagent bottle 1 is 300mL to 1000mL, and is mainly used to store dewaxing agent, detergent, deionized water, anhydrous ethanol, gradient ethanol, and post-hybridization washing solution.

[0030] The second reagent liquid path system includes at least one second reagent supply unit, which includes a second reagent bottle 12, a pipette tip 9, a microinjection pump 7 and a plurality of disposable pipette tips 10 arranged on the equipment reaction platform 4. The tail of the pipette tip 9 is connected to the microinjection pump 7. The pipette tip 9 is used to absorb the disposable pipette tip 10 and insert the disposable pipette tip 10 into the second reagent bottle 12, and the reagent in the second reagent bottle 12 is pumped into the corresponding disposable pipette tip 10 through the microinjection pump 7. The second reagent liquid path system removes the disposable pipette tip 10 from the second reagent bottle 12 and moves it to the reaction tank 5 to pump out the second reagent. After the reaction is completed, the reagent in the reaction tank 5 is emptied.

[0031] In one embodiment, at least 10 second reagent supply units are provided, which are composed of 10 second reagent bottles 12, 96 disposable pipette tips 10 and a microinjection pump 7. The 96 disposable pipette tips 10 are placed on a 96-well plate 11 of the device reaction platform 4 and need to be replaced each time a different type of reagent is taken. The reaction tank 5 is centrally arranged on the device reaction platform 4. The volume of the second reagent bottle 12 is 0.5mL to 10mL, and is mainly used to store permeabilization agent, sealing oil, diluent, FISH probe, and DAPI.

[0032] The third reagent liquid path system includes at least one third reagent supply unit, which includes a third reagent air pump 35, a third reagent bottle 31, a third reagent ventilation pipe 34, a third reagent infusion pipe 33 and a third reagent nozzle 32. The third reagent air pump 35 is connected to the third reagent bottle 31 through the third reagent ventilation pipe 34, and the third reagent bottle 31 is connected to the third reagent nozzle 32 through the third reagent infusion pipe 33.

[0033] The third reagent air pump 35 is used to introduce a certain amount of air into the third reagent bottle 31. When the air pressure in the third reagent bottle 31 is greater than the atmospheric pressure outside the bottle, the reagent in the third reagent bottle 31 will be squeezed out through the liquid outlet of the third reagent bottle 31, and then introduced into the reaction tank 5 through the third reagent infusion tube 33 and the third reagent nozzle 32. After the reaction is completed, the reagent in the reaction tank 5 is emptied. The volume of the third reagent bottle 31 is 20mL to 200mL, and it is mainly used to store protease solution.

[0034] The emptying of the reagent in the reaction tank 5 is completed by the reagent emptying module of the reaction tank 5. The reagent emptying module of the reaction tank 5 includes a strip-shaped air nozzle 21, a reagent emptying module air pump 22 and a reagent emptying module air pipe 23, and the reagent emptying module air pipe 23 connects the reagent emptying module air pump 22 and the strip-shaped air nozzle 21. The reaction tank 5 emptying module emits a strip-shaped airflow through the strip-shaped air nozzle 21 to flush the residual reagent in the reaction tank 5, and empties all the reagents in the reaction tank 5 under the synergistic effect of the displacement platform.

[0035] Among them, one waste liquid diversion pipe 6 is provided, and one waste liquid barrel is provided.

[0036] The control accuracy of the first reagent liquid circuit system can reach ±0.5mL, the control accuracy of the second reagent liquid circuit system can reach ±1μL, and the control accuracy of the third reagent liquid circuit system can reach ±0.2mL. The first reagent liquid circuit system is for reagents with large consumption and low price, the second reagent liquid circuit system is for reagents with small consumption and high price, and the third reagent liquid circuit system is for reagents with moderate consumption and price, so that the liquid circuit system can achieve accurate control of the amount of various reagents, save reagent consumption, and reduce the cost of FISH experiments.

[0037] The present invention also provides a fully automatic multi-chip independent FISH staining device, which includes the liquid path system and controller for the fully automatic multi-chip independent FISH staining device as described above, and the controller is connected to the liquid path system.

[0038] It will be easily understood by those skilled in the art that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A liquid path system for a fully automatic multi-chip independent FISH staining device, characterized in that: The liquid circuit system includes a first reagent liquid circuit system, a second reagent liquid circuit system, a third reagent liquid circuit system and an independent sample reaction chamber, the independent sample reaction chamber includes a plurality of independent reaction slots and a sample clamping chamber, and the plurality of reaction slots are arranged in parallel; a sample clamping chamber is arranged in each reaction slot, and the sample clamping chamber is used to clamp sample glass slides, and the first reagent liquid circuit system, the second reagent liquid circuit system and the third reagent liquid circuit system are used to independently add reagents to the sample clamping chamber.

2. The liquid path system for a fully automatic multi-chip independent FISH staining device according to claim 1, characterized in that: The first reagent liquid path system includes at least one first reagent supply unit, the first reagent supply unit includes a first reagent bottle, a peristaltic pump and a flow meter, the peristaltic pump is connected to the first reagent bottle and the flow meter, the flow meter is connected to a first reagent discharge port through a pipeline, and the first reagent discharge port is used to introduce the reagent from the first reagent supply unit into the corresponding reaction tank.

3. The liquid path system for a fully automatic multi-chip independent FISH staining device according to claim 2, characterized in that: At least six first reagent supply units are provided; the volume of the first reagent bottle is 300mL-1000mL.

4. The liquid path system for a fully automatic multi-chip independent FISH staining device according to claim 1, characterized in that: The independent sample reaction chamber is provided with at least 10 reaction slots, and the reaction slots are arranged in parallel.

5. The liquid path system for a fully automatic multi-chip independent FISH staining device according to claim 1, characterized in that: The second reagent liquid path system includes at least one second reagent supply unit, which includes a second reagent bottle, a pipette tip, a microinjection pump and a plurality of disposable pipette tips arranged on the equipment reaction platform, the tail of the pipette tip is connected to the microinjection pump, the pipette tip is used to suck the disposable pipette tip and insert the disposable pipette tip into the second reagent bottle, and the reagent in the second reagent bottle is pumped into the corresponding disposable pipette tip through the microinjection pump; the second reagent liquid path system pumps out the second reagent by removing the disposable pipette tip from the second reagent bottle and moving it into the reaction tank.

6. The liquid path system for a fully automatic multi-chip independent FISH staining device according to claim 5, characterized in that: At least 10 second reagent supply units are provided; the volume of the second reagent bottle is 0.5 mL to 10 mL.

7. The liquid path system for a fully automatic multi-chip independent FISH staining device according to claim 1, characterized in that: The third reagent liquid path system includes at least one third reagent supply unit, and the third reagent supply unit includes a third reagent air pump, a third reagent bottle, a third reagent ventilation tube, a third reagent infusion tube and a third reagent nozzle. The third reagent air pump is connected to the third reagent bottle through the third reagent ventilation tube, and the third reagent bottle is connected to the third reagent nozzle through the third reagent infusion tube.

8. The liquid path system for a fully automatic multi-chip independent FISH staining device according to claim 7, characterized in that: The volume of the third reagent bottle is 20 mL to 200 mL; the reaction tanks are centrally arranged on the equipment reaction platform.

9. The liquid path system for a fully automatic multi-chip independent FISH staining device according to any one of claims 1 to 8, characterized in that: The emptying of the reagent in the reaction tank is completed by the reaction tank reagent emptying module; the reaction tank reagent emptying module includes a strip air nozzle, a reagent emptying module air pump and a reagent emptying module air pipe, and the reagent emptying module air pipe connects the reagent emptying module air pump and the strip air nozzle; the reaction tank emptying module emits a strip airflow through the strip air nozzle to flush the residual reagent in the reaction tank.

10. A fully automatic multi-slice independent FISH staining device, characterized by: It comprises a liquid path system and a controller for a fully automatic multi-chip independent FISH staining device as described in any one of claims 1 to 9, wherein the controller is connected to the liquid path system.

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