Patent Application
20220205996
The present application is based on and claims the priority benefits of China application No. 202011639404.7, filed on Dec. 31, 2020. The entirety of the above-mentioned patent application is hereby incorporated by reference herein and made a part of this specification.
Technical Field
The invention relates to the technical field of protein immunoblotting analysis, in particular to an immunoblotting instrument and a control method thereof.
Immunoblotting methods are commonly used to identify certain proteins, can make qualitative and semi-quantitative analysis of protein, and can be extensively used in the fields of life science research and laboratory medicine, such as tumor marker detection and pathogenic microorganism detection. This method transfers the target protein separated by electrophoresis from the gel to a solid support (such as NC membrane or PVDF membrane), and then hybridizes to the specific ligand of the known protein (such as antibody, etc.). Through an enzyme system after color development, the target protein is detected. The immunodetection part successively includes five processes of blocking, primary antibody hybridization, washing, secondary antibody hybridization, and washing, involving multiple steps of liquid addition, liquid aspiration, incubation, and cleaning.
In order to perform the above steps, some immunoblotting instruments can automatically perform the above steps of liquid addition, liquid aspiration, incubation and so on according to a set program. However, by analyzing various types of immunoblotting instruments of the prior art, the inventors found that none of the current immunoblotting instruments can precisely control the incubation reaction conditions of immunoblotting membranes, and the immunoblotting membranes have different effects of the contact reaction with reagents under different environmental conditions, which means that different incubation reaction environments will cause different test results, and ultimately reduce the reliability of the test results.
For the problem that the reliability of the detection result of the immunoblotting instrument needs to be further improved in practical application, it is an object of the present application to provide an immunoblotting instrument, which through precise control of immunoblotting membrane incubation and reaction environment condition, can ensure the reliability of detection result. Based on the immunoblotting instrument, another objective of the application is to provide a control method for controlling the immunoblotting instrument. The specific scheme is as follows: an immunoblotting instrument comprises an immunoblotting instrument body, and an immunoblotting instrument control device, an incubation device and a liquid feeding and sucking device that are provided on the immunoblotting instrument body, and further comprises a temperature control device for controlling the incubation environment temperature of the immunoblotting membrane.
Through the above-mentioned technical scheme, the incubation environment temperature in the incubation device can be controlled so as to ensure that the reaction between the immunoblotting membrane and the specific protein in the incubation device can occur under the set environment temperature condition, and the accuracy of the detection result is ensured. Meanwhile, the set temperature can also improve the tightness of binding of the immunoblotting membrane and the specific protein, so that the whole detection efficiency can be improved, and the detection difficulty of the target protein is reduced.
Further, the incubation device comprises a tray rack provided on the immunoblotting instrument body, and a tray detachably mounted on the tray rack for containing the immunoblotting membrane and the reagent; the temperature control device comprises a refrigerator, a refrigeration plate provided with a plurality of refrigeration channels, and a refrigeration conduit communicated with the refrigerator and the refrigeration plate to form a refrigeration circuit, wherein the refrigeration conduit is provided with a liquid pump for driving a cooling liquid in the refrigeration conduit to flow; the refrigerator and the liquid pump are electrically connected to the immunoblotting instrument control device, receive the signal for refrigeration control output by the immunoblotting instrument control device and control the refrigeration power and the flow rate of the cooling liquid; and the refrigeration plate is made of heat conducting metal and is provided at a position on the tray rack supporting the tray.
Through the above technical scheme, the temperature of the tray for storing the immunoblotting membrane can be accurately controlled, so that the incubation reaction temperature of the immunoblotting membrane can be controlled, and the reliability and accuracy of the detection result are ensured.
Further, the liquid feeding and sucking device comprises a holder rack for holding the reagent bottle, a liquid injection conduit for injecting the reagent in the reagent bottle into the tray, a liquid suction conduit for discharging the reagent or waste liquid in the tray, and a peristaltic pump provided on the liquid injection conduit and the liquid suction conduit;
Through the above technical scheme, the temperature of the reagent injected into the tray can be adjusted, so that the incubation temperature in the tray is always kept at a set value, and the incubation environment condition of the immunoblotting membrane is kept stable.
Further, the holder rack is provided with a plurality of holding cups matched with the reagent bottles in shape and size, the adjustment member comprises a temperature adjusting conduit or temperature adjusting chamber that is provided around the outer side of the holding cups, filled with a heat-conducting medium and communicated with the refrigerator, and a conducting pump is provided between the temperature adjusting conduit or temperature adjusting chamber and the refrigerator;
Through the above technical scheme, cold generated by the refrigerator is transferred to the holding cup through the temperature adjusting conduit or temperature adjusting chamber, so that the temperature of reagents in the reagent bottle is maintained within a reasonable range.
Further, the temperature adjusting conduit or temperature adjusting chamber is in communication with the refrigeration conduit to jointly form the refrigeration circuit.
Through the above technical scheme, the temperature of the reagent injected into the tray is consistent with the temperature of the existing reagent in the tray. It is ensured that great mutation of the incubation environment condition in the tray would not happen due to the injection of the reagent.
Further, the temperature control device further comprises a temperature sensor for measuring the temperature of the refrigeration plate that is in signal connection with the control device, and the immunoblotting instrument control device receives a temperature detection signal output by the temperature sensor and adjusts the action of the refrigerator and/or the liquid pump
Through the above technical scheme, the incubation temperature can be monitored and adjusted in real time according to the actual temperature in the tray, and the temperature is ensured to be in a proper range.
Further, the tray rack is movably provided on the immunoblotting instrument body through a mounting seat, and the immunoblotting instrument body is provided with an oscillating assembly for driving the tray rack to reciprocatedly vibrate.
Through the above technical scheme, when the immunoblotting membrane is placed in the tray for incubation, the vibration of the tray rack enables the immunoblotting membrane to be fully contacted with the reagent, so that the reaction efficiency is improved.
Further, the immunoblotting instrument control device comprises a power supply, a control main board and a man-machine interaction assembly. The control main board is provided with a memory and a processor, the memory is loaded with a set program for step execution, and the processor receives and responds to the input of the man-machine interaction assembly to control the actions of the incubation device, the liquid feeding and sucking device and the temperature control device based on the set step execution program in the memory.
Further, the refrigeration conduit comprises a main refrigeration conduit and at least one auxiliary refrigeration conduit that are connected in parallel with each other, two ends of the main refrigeration conduit and of the auxiliary refrigeration conduit are provided with an electrically-controlled multi-way valve electrically connected to the immunoblotting instrument control device; and
Through the above technical scheme, after reagent is transmitted through different refrigeration conduits, the temperatures of regent input into the tray can be different, so that different refrigeration conduits can be selected as transmission pipelines of the cooling liquid according to needs, and accurate control of the temperatures in the tray is guaranteed.
Based on the immunoblotting instrument, the invention also provides a control method for controlling the immunoblotting instrument comprising the following steps of:
Compared with the prior art, the beneficial effects of the present invention are as follows:
Reference numerals: 1. Immunoblotting instrument Control device; 2. Incubation device; 3. Liquid feeding and sucking device; 4. Temperature control device; 5. Tray rack; 6. Tray; 7. Mounting groove; 8. Holding groove; 9. Reagent bottle; 10. Holder rack; 11.Liquid injection conduit; 12. Liquid suction conduit; 13. Peristaltic pump; 14. Holding cup; 15. Hanging arm; 16. Passing hole; 17. Refrigerator; 18. Refrigeration plate; 19. Refrigeration conduit; 20. Liquid pump; 22. Control main board; 23. Man-machine interaction assembly; 24. Immunoblotting instrument housing; 25. Touch display screen; 26. Temperature adjusting chamber; 27. Heat conducting medium; 28. Conducting pump; 29. Mounting seat; 30. Connecting plate; 31. Drive motor; 32. Via hole; 33. Cam; 34. Roller shaft; 35. Driven gear; 36. Drive gear; 37. Synchronous belt ; 38. Vertical transmission rod.
The invention is described in further detail below in conjunction with the examples and drawings, but embodiments of the invention are not limited thereto.
An immunoblotting instrument, as shown in
In an embodiment of the present application, the incubation device 2 comprises a tray rack 5 provided on the immunoblotting instrument body, and a tray 6 detachably mounted on the tray rack 5 for storing the immunoblotting membrane and the reagent as shown. As shown in
In the embodiment, the tray 6 is made of transparent plastics and is convenient to use and detect; and the tray 6 is disposable so as to avoid the cross contamination.
In order to make the reagent injected into the tray 6 sufficiently contact and react with the immunoblotting membrane placed in the tray 6, an oscillating assembly is provided between the tray rack 5 and the top surface of the immunoblotting instrument housing 24.
Specifically, in connection with
As shown in
A vertical transmission rod 38 is connected between the position close to the edge of the cam 33 and the tray rack 5. The two ends of the vertical transmission rod 38 are respectively hinged with the cam 33 and the tray rack 5. When the cam 33 rotates, the vertical transmission rod drives the tray rack 5 to reciprocatedly vibrate with the mounting seat 29 as a supporting point. In some embodiments, in order to increase the amplitude of the tray rack 5, the connecting plate 30 is provided at the bottom of the pallet rack 5 at a position near its long side.
In an embodiment of the present application, in order to inject the reagent in the reagent bottle 9 into the tray 6, the liquid feeding and sucking device 3 comprises a holding rack 10 for holding the reagent bottle 9, a liquid injection conduit 11 for injecting the reagent in the reagent bottle 9 into the tray 6, a liquid suction conduit 12 for discharging the reagent or waste liquid in the tray 6, and a peristaltic pump 13 provided on the liquid injection conduit 11 and the liquid suction conduit 12. In connection with
In order to smoothly introduce the reagent in the reagent bottle 9 into the tray 6, an opening at an end of the liquid injection conduit 11 far away from the reagent bottle 9 extends to the position above the tray 6. In order to fix the above liquid injection conduit 11, an inverted L-shaped hanging arm 15 is provided on the tray rack 5. An end of the hanging arm 15 is hinged with the tray rack 5, and the other end thereof is a free end and extends to the position right above the tray 6. The hanging arm 15 is locked with the tray rack 5 through a locking piece, such as a clamping block, a bolt and so on. A passing hole 16 for clamping the conduit is provided on the free end of the hanging arm 15, so that the opening of the liquid injection conduit 11 is stably positioned right above the tray 6, and the reagent in the reagent bottle 9 can be directly injected into the tray 6 after being pumped by the peristaltic pump 13.
In an embodiment of the present application, a plurality of reagent bottles 9 are provided, and respectively store a reagent for reaction, a cleaning liquid for cleaning the tray 6, and a waste liquid. Correspondingly, the number of the peristaltic pumps 13 and the number of the liquid injection conduits 11 and the number of the liquid suction conduits 12 are multiple. The above-mentioned liquid injection conduits 11 and the liquid suction conduits 12 each pass through the interior of the immunoblotting instrument housing 24. The peristaltic pumps 13 are all electrically connected to the immunoblotting instrument control device 1, receive and respond to a control signal output from the immunoblotting instrument control device 1 to act. For example, a reagent a in a reagent bottle 9a is injected into a first tray 6 by a liquid injection conduit 11a, a reagent b in a reagent bottle 9b is injected into the first tray 6 by a liquid injection conduit 11b, and waste liquid in the first tray 6 is sucked into a waste liquid collecting bottle or a drain pipe by a liquid suction conduit 12.
In order to maintain the temperature in the tray 6 in a proper range, ensure the activity of protein and ensure that the incubation and reaction can occur efficiently and smoothly, the immunoblotting instrument in the example also comprises a temperature control device 4 for controlling the incubation environment temperature of the immunoblotting membrane.
The temperature control device 4 comprises a refrigerator 17 provided in the immunoblotting instrument housing 24, a refrigeration plate 18 provided with a plurality of refrigeration channels, and a refrigeration conduit 19 communicated with the refrigerator 17 and the refrigeration plate 18 to form a refrigeration circuit. The refrigeration conduit 19 is provided with a liquid pump 20 for driving the cooling liquid in the refrigeration conduit 19 to flow, which may be configured as a peristaltic pump. The refrigerator 17 and the liquid pump 20 are electrically connected to the immunoblotting instrument control device 1, receive the signal for refrigeration control output from the immunoblotting instrument control device 1 to control the refrigeration power of the refrigerator 17 and the pumping rate of the liquid pump 20, so as to control the temperature and the flow rate of the cooling liquid. In an embodiment of the present application, the refrigeration plate 18 is made of a heat-conducting metal, such as aluminum. As shown in
In some embodiments, the refrigeration conduit 19 may be configured as a main refrigeration conduit and at least one auxiliary refrigeration conduit that are connected in parallel with each other. The two ends of the main refrigeration conduit and of the auxiliary refrigeration conduit are provided with an electrically-controlled multi-way valve electrically connected to the immunoblotting instrument control device 1. Based on the above settings, different refrigeration pipelines may be selected to be connected to the refrigeration circuit, and a plurality of the transmission circuit of the cooling liquid may be provided. The heat conductivity coefficients of the main refrigeration conduit and the auxiliary refrigeration conduit are different from each other. For example, the outer side of the main refrigeration conduit may be coated with an insulating layer, such as a heat insulation rubber felt layer. The outer side of the auxiliary refrigeration conduit may be coated with a heat-radiating layer, such as a heat-radiating film; or the auxiliary refrigeration conduit is directly provided as a corrugated pipe. As such, after the reagent is transmitted through different refrigeration conduits 19, the temperature of the cooling liquid that is finally transferred to the tray 6 from different refrigeration conduits 19 will be different, so that different refrigeration conduits 19 (main refrigeration conduit and/or the auxiliary refrigeration duct) can be selected according to the need as the transmission pipeline of the cooling liquid, and the cooling liquid with different temperatures can be adjusted to achieve more diverse and accurate temperatures, so as to realize the precise control of the environment temperature in the tray 6.
Since different reagents need to be injected into the tray 6 at different time according to the preset program or the control signal input through the man-machine interaction assembly during the entire incubation reaction process, in order to avoid sudden changes in the temperature in the tray 6 after the reagents are injected, the temperature control device 4 also includes an adjustment member for adjusting and controlling the environment temperature of the holding rack 10 for the reagent bottle 9.
As shown in
By adopting the technical scheme, the temperature of the reagent in the reagent bottle 9 placed in the holding cup 14 can be maintained within a set range, so that the temperature in the tray 6 cannot change suddenly when the reagent is injected into the tray 6.
In another embodiment, refrigeration of the holding cup 14 may be achieved by winding a temperature adjusting conduit communicating with the refrigerator 17 around the outer side of the holding cup 14.
In some embodiments, the refrigeration conduit 19, the refrigerator 17, the liquid pump 20 and the temperature adjusting chamber 26 in the present application jointly form a refrigeration circuit, that is, the temperature adjustment chamber 26 and the refrigeration plate 18 are connected through the refrigeration conduit 19 and the liquid pump 20. In this way, the temperature of the reagent injected into the tray 6 is consistent with that of the existing reagent in the tray 6, and the incubation environment condition in the tray 6 is ensured not to be greatly changed due to the injection of the reagent.
To accurately control the temperature of the reagent in the tray 6, the temperature control device 4 in the present application further comprises a temperature sensor for measuring the temperature of the refrigeration plate 18. The temperature sensor is electrically connected to the immunoblotting instrument control device 1, collects the temperature on the refrigeration plate 18 and outputs the temperature detection signal to the immunoblotting instrument control device 1. Based on the temperature detection signal, the immunoblotting instrument control device 1 adjusts the action of the refrigerator 17 and/or the liquid pump 20. For example, when the temperature in the tray 6 is too high, the refrigeration power of the refrigerator 17 or the pumping power of the liquid pump 20 is increased, so that the temperature of the cooling liquid is reduced or the flow rate of the cooling liquid is accelerated, the heat in the tray 6 is taken away more quickly to realize the accurate control of the temperature, and the activity of the target protein in the tray 6 is ensured.
As shown in
It should be noted that the plurality of liquid feeding and sucking devices 3 and temperature control devices 4 are independently provided to provide different incubation and reaction temperatures to the plurality of trays 6.
Based on the immunoblotting instrument, the invention also provides a control method for controlling the immunoblotting instrument, as shown in
In the above step S100, based on the characteristics of various target proteins to be detected, a corresponding detection program is stored in advance in the memory of the immunoblotting instrument control device 1, such as the temperature in the tray 6, the time for filling reagent, the time for reaction and the time for feeding the liquid, and then according to the above-mentioned steps a correspondent immunoassay control table is generated. The above-mentioned immunoassay control table is actually a control code that can be directly stored in the single-chip microcomputer or a memory chip connected with it.
In the step S200, the operator communicates different liquid injection conduits 11 and liquid suction conduits 12 with corresponding reagent bottles 9 before detection, and then detects whether the instrument can be controlled normally or not. When the detection is completed, the instrument is started to begin detection, The immunoblotting instrument control device 1 in the instrument can control the operation of the peristaltic pump 13 according to a set control program so as to regularly and quantitatively extract and inject the reagent into the tray 6, and suck the reagent or waste liquid out of the tray 6 at a set time; meanwhile, it can stabilize the reaction temperature of the whole process, so that the normal reaction is ensured; manual intervention is not needed in the detection process, so that the detection can be efficiently and automatically finished with a high efficiency. Since the reaction environment is consistent, the reliability of the detection result is improved.
Based on the above-mentioned steps, as detailed, the above-mentioned control method for controlling an immunoblotting instrument further includes the following steps:
The foregoing description is merely preferred embodiments of the invention, The scope of the invention is not limited to the embodiments described above. It should be noted that modifications and amendments, which fall within the scope of the present invention, will also be considered to those of ordinary skill in the art without departing from the principles of the present invention as falling within the scope of the invention.
| Number | Date | Country | Kind |
|---|---|---|---|
| 202011639404.7 | Dec 2020 | CN | national |
