A method and system for sample loading control for PCR reaction plate making

Abstract

This invention discloses a sample loading control method and system for PCR reaction plate preparation, relating to the field of PCR reaction plate preparation technology. The method includes: acquiring well position information of each spray needle in a spraying device that needs to be sprayed onto the reaction plate; generating a motion control program for the current spray needle based on the well position information and a preset movement speed; calculating the well spacing between two adjacent wells that need to be sprayed in the reaction plate, based on the well position information; calculating the PWM period for the spraying operation of the spray needle based on the well spacing, well position information, and preset movement speed; generating a spraying control program for the current spray needle based on the PWM period; and simultaneously executing the motion control program and the spraying control program. This invention significantly shortens the preparation time of PCR reaction plates, reduces the preparation cost of PCR reaction plates, and improves the accuracy of PCR experimental results.

Description

Technical Field

[0001] This invention relates to the field of PCR reaction plate preparation technology, and in particular to a sample addition control method and system for PCR reaction plate preparation. Background Technology

[0002] In current PCR experiments, reagent spraying on reaction plates involves a row / column of spray needles simultaneously targeting different wells on the plate. Each row / column of spray needles contains the same reagent. Since the spraying positions of the needles are pre-set to be fixed, it is impossible to spray the reagent onto other wells. However, it is necessary to add different reagents multiple times to the same well, and the addition of each well may not be the same. In this case, it is often necessary to complete the addition task in batches through multiple rounds of addition. The resulting problem is that it is necessary to increase the number of times the spray needles need to be cleaned, resulting in the loss of cleaning solution. Improper cleaning can also cause deviations in the PCR reaction system, affecting the PCR experimental results, and also greatly increasing the preparation time of the PCR reaction plate.

[0003] Therefore, a sample loading control method and system for PCR reaction plate preparation was developed to solve the above problems. Summary of the Invention

[0004] This invention proposes a sample loading control method and system for PCR reaction plate preparation, in order to solve the problems of long preparation time, high cost, and susceptibility to accuracy issues in existing PCR reaction plate preparation.

[0005] The present invention achieves the above objectives through the following technical solutions: This invention provides a sample loading control method for PCR reaction plate preparation, comprising: Obtain the hole position information of each spray needle in the spray device on the reaction plate that needs to be sprayed. The spray device includes multiple spray needles, each of which corresponds to a sample type. Based on the orifice information and the preset movement speed, a motion control program for the current spray needle is generated; the motion control program is used to control the spray needle to pass through at least each orifice in the orifice information on the reaction plate at the preset movement speed. Based on the hole position information, calculate the hole spacing between two adjacent holes that need to be sprayed in the reaction plate that the current spray needle passes through in sequence. The PWM cycle for the liquid spraying operation of the spray needle is calculated based on the hole spacing, hole position information and preset movement speed. Based on the PWM cycle, a spray control program for the current spray needle is generated. The spray control program is used to control the current spray needle to add samples to the corresponding hole position according to the PWM cycle. Simultaneously execute motion control program and liquid spraying control program.

[0006] Furthermore, before executing the motion control program and the liquid spraying control program, the following are also included: Initialize the virtual orifice position, which is the first orifice position in each column on the reaction plate; or, the virtual orifice position is located outside the first orifice in each column on the reaction plate where liquid spraying begins, and is one orifice spacing away from the first orifice. The initial virtual orifice position is set as the starting point for the movement of the injection needle, as well as the reference point for timing the start of the PWM cycle and motion control.

[0007] Furthermore, the PWM cycle for the liquid spraying operation of the spray needle is calculated based on the hole spacing, hole position information, and preset movement speed, including: When only one hole spacing is calculated based on the hole position information, a PWM cycle is independently calculated based on the preset motion speed and the hole spacing, according to the preset motion relationship. When multiple hole spacings are calculated based on hole position information, different PWM cycles corresponding to each hole spacing are independently calculated based on a preset motion relationship, according to a preset motion speed and each different hole spacing. The preset motion relationship is as follows: Hole spacing = movement speed × PWM cycle.

[0008] Furthermore, it also includes optimizing the movement speed of the injection needle based on the orifice position information, including: Based on the orifice information, the movement trajectory of the spray needle on the reaction plate is divided into a continuous spray segment and a continuous non-spray segment, both of which include the case of a single orifice. The spray needle moves at a first speed in the continuous spray segment; The spray needle moves at a second speed in the continuous non-spraying section; The second speed is equal to or greater than the first speed.

[0009] If there are multiple orifice spacings in the motion trajectory, and the same motion speed is used in different sections, if the focus is on shortening the time, the accuracy of the spray cannot be guaranteed. If the focus is on the accuracy of the spray, this will cause redundancy in the total sample loading time and reduce the spray efficiency. Therefore, setting different motion speeds for different sections can achieve a balance between spray accuracy and efficiency.

[0010] Furthermore, the motion control program controls the spray needle to move from the first hole in the current column of the reaction plate that needs to be sprayed to the last hole in the current column that needs to be sprayed, and then from the last hole in the next column to the first hole in that column, and so on. The spray needle moves back and forth along the reaction plate column by column until it passes through every hole in the hole position information.

[0011] Furthermore, the motion control program is used to control the spray needle to pass through all the holes on the reaction plate at a preset speed.

[0012] Furthermore, the motion control program and the liquid spraying control program are executed simultaneously, including: Motion control commands are generated based on the motion control program; The drive motor used to drive the spray needle is controlled according to motion control commands; Generate spray control commands based on the spray control program; The spray valve of the spray needle is controlled according to the spray control command.

[0013] Furthermore, a series of spray needles are integrated into a single structure and driven by the same drive motor.

[0014] The present invention also provides a system for the sample loading control method for PCR reaction plate preparation, characterized in that it comprises: The information acquisition module is used to acquire the hole position information of each spray needle in the spray device on the reaction plate that needs to be sprayed. The spray device includes multiple spray needles, each of which corresponds to a sample type. The first generation module is used to generate a motion control program for the current spray needle based on the orifice information and the preset motion speed; the motion control program is used to control the spray needle to pass through at least each orifice in the orifice information on the reaction plate at the preset motion speed. The first calculation module is used to calculate the hole spacing between two adjacent holes that need to be sprayed in the reaction plate through which the current spray needle passes, based on the hole position information. The second calculation module is used to calculate the PWM cycle of the liquid spraying operation of the liquid spraying needle based on the hole spacing, hole position information and preset movement speed. The second generation module is used to generate a liquid spraying control program for the current liquid spraying needle according to the PWM cycle. The liquid spraying control program is used to control the current liquid spraying needle to add samples to the corresponding hole position according to the PWM cycle. An execution control module is used to simultaneously execute a motion control program and a liquid spraying control program.

[0015] The beneficial effects of this invention are as follows: The sample loading control method and system proposed in this invention for PCR reaction plate preparation greatly shortens the preparation time of PCR reaction plates, reduces the preparation cost of PCR reaction plates, and improves the accuracy of PCR experimental results. Attached Figure Description

[0016] Figure 1 This is a diagram illustrating the sample loading process used in the preparation of PCR reaction plates in this application. Detailed Implementation

[0017] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. The components of the embodiments of the present invention described and shown in the accompanying drawings can generally be arranged and designed in various different configurations.

[0018] Therefore, the following detailed description of the embodiments of the invention provided in the accompanying drawings is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the invention without inventive effort are within the scope of protection of the invention.

[0019] It should be noted that similar labels and letters in the following figures indicate similar items. Therefore, once an item is defined in one figure, it does not need to be further defined and explained in subsequent figures.

[0020] Furthermore, the terms "first," "second," etc., are used only to distinguish descriptions and should not be interpreted as indicating or implying relative importance.

[0021] The specific embodiments of the present invention will now be described in detail with reference to the accompanying drawings.

[0022] like Figure 1 As shown, a sample loading control method and system for PCR reaction plate preparation includes: Obtain the hole position information of each spray needle in the spray device on the reaction plate that needs to be sprayed. The spray device includes multiple spray needles, each of which corresponds to a sample type. Based on the orifice information and the preset movement speed, a motion control program for the current spray needle is generated; the motion control program is used to control the spray needle to pass through at least each orifice in the orifice information on the reaction plate at the preset movement speed. Based on the hole position information, calculate the hole spacing between two adjacent holes that need to be sprayed in the reaction plate that the current spray needle passes through in sequence. The PWM cycle for the liquid spraying operation of the spray needle is calculated based on the hole spacing, hole position information and preset movement speed. Based on the PWM cycle, a spray control program for the current spray needle is generated. The spray control program is used to control the current spray needle to add samples to the corresponding hole position according to the PWM cycle. Simultaneously execute motion control program and liquid spraying control program.

[0023] In one embodiment, before executing the motion control program and the liquid spraying control program, the method further includes: Initialize the virtual orifice position, which is the first orifice position in each column on the reaction plate; or, the virtual orifice position is located outside the first orifice in each column on the reaction plate where liquid spraying begins, and is one orifice spacing away from the first orifice. The initial virtual orifice position is set as the starting point for the movement of the injection needle, as well as the reference point for timing the start of the PWM cycle and motion control.

[0024] In one embodiment, the PWM period for the liquid spraying operation of the spray needle is calculated based on the hole spacing, hole position information, and a preset movement speed, including: When only one hole spacing is calculated based on the hole position information, a PWM cycle is independently calculated based on the preset motion speed and the hole spacing, according to the preset motion relationship. When multiple hole spacings are calculated based on hole position information, different PWM cycles corresponding to each hole spacing are independently calculated based on a preset motion relationship, according to a preset motion speed and each different hole spacing. The preset motion relationship is as follows: Hole spacing = movement speed × PWM cycle.

[0025] In one embodiment, the method further includes optimizing the movement speed of the injection needle based on the orifice location information, including: Based on the orifice information, the movement trajectory of the spray needle on the reaction plate is divided into a continuous spray segment and a continuous non-spray segment, both of which include the case of a single orifice. The spray needle moves at a first speed in the continuous spray segment; The spray needle moves at a second speed in the continuous non-spraying section; The second speed is equal to or greater than the first speed.

[0026] If there are multiple orifice spacings in the motion trajectory, and the same motion speed is used in different sections, if the focus is on shortening the time, the accuracy of the spray cannot be guaranteed. If the focus is on the accuracy of the spray, this will cause redundancy in the total sample loading time and reduce the spray efficiency. Therefore, setting different motion speeds for different sections can achieve a balance between spray accuracy and efficiency.

[0027] In one embodiment, the motion control program controls the spray needle to move from the first hole in the current column of the reaction plate that needs to be sprayed to the last hole in the current column that needs to be sprayed, and then from the last hole in the next column to the first hole in that column, and so on. The spray needle moves back and forth along the reaction plate column by column until the spray needle passes through every hole in the hole position information.

[0028] In one embodiment, the motion control program is used to control the spray needle to pass through all the holes on the reaction plate at a preset motion speed.

[0029] In one embodiment, the motion control program and the liquid spraying control program are executed simultaneously, including: Motion control commands are generated based on the motion control program; The drive motor used to drive the spray needle is controlled according to motion control commands; Generate spray control commands based on the spray control program; The spray valve of the spray needle is controlled according to the spray control command.

[0030] In one embodiment, a row of spray needles is integrated into a single structure and driven by a single drive motor. Since the control of each spray needle during spraying is located in a different thread, the logic control between the spray needles is decoupled, and they do not interfere with each other. Therefore, this method can accommodate any number of needles for sample application. Furthermore, different combinations of motion speed and PWM cycle can be adjusted according to the hole spacing of different board specifications, thus enabling adaptation to reaction boards of different sizes.

[0031] The present invention also provides a system for the sample loading control method for PCR reaction plate preparation, characterized in that it comprises: Obtain the hole position information of each spray needle in the spray device on the reaction plate that needs to be sprayed. The spray device includes multiple spray needles, each of which corresponds to a sample type. The first generation module is used to generate a motion control program for the current spray needle based on the orifice information and the preset motion speed; the motion control program is used to control the spray needle to pass through at least each orifice in the orifice information on the reaction plate at the preset motion speed. The first calculation module is used to calculate the hole spacing between two adjacent holes that need to be sprayed in the reaction plate through which the current spray needle passes, based on the hole position information. The second calculation module is used to calculate the PWM cycle of the liquid spraying operation of the liquid spraying needle based on the hole spacing, hole position information and preset movement speed. The second generation module is used to generate a liquid spraying control program for the current liquid spraying needle according to the PWM cycle. The liquid spraying control program is used to control the current liquid spraying needle to add samples to the corresponding hole position according to the PWM cycle. An acquisition module is used to simultaneously execute a motion control program and a liquid spraying control program.

[0032] The implementation process of this invention in a specific application is illustrated below: like Figure 1Multiple needles (DJ1, DJ2, DJ3, DJ4) are arranged in a row to form a spray device. When adding samples, the spray device moves from the first well in a row that needs to be added to the last well, ensuring that the tip of each needle passes over each well and then returns from the other side to pass over the next row of wells. This process is repeated over the entire plate of wells. Figure 1 The barcode area contains the identification code for the reaction plate information. The reaction hole is the hole position. For odd-numbered columns, the spraying direction is from top to bottom, and for even-numbered columns, the spraying direction is from bottom to top.

[0033] Before the sample dispensing process begins, the dispensing position information of each nozzle on the reaction plate is transmitted to the processor of the control device. The CPU then assigns a control thread to each nozzle to process the dispensing information.

[0034] The entire spraying action is divided into minimum control units, with columns as the unit. A virtual hole position is determined outside the first hole in each column where spraying begins. Based on the spraying information, the hole position to be sprayed by each spray needle in the current column is determined. Holes with the same spray needle are grouped together, and the following information is calculated for each group: Using virtual orifices as a reference, the time to reach the first spray orifice is calculated. Consecutive spray orifices are grouped into a single spray group, and the time to complete one spray group is calculated. Similarly, consecutive non-spraying orifices are grouped into a single non-spraying group, and the time to complete one non-spraying group is calculated. This converts the spray information of a column into a set of time information. Using the virtual point as a reference, timing begins when the needle passes the virtual point, sequentially timing each time segment within the time information group. The duration of each time segment is the spraying and closing time of the spray needle. If all orifices in a column require spraying, the spray needle will remain open, and the spraying duration is the PWM (Pulse Width Modulation) duration. Within each spraying time, the calculated PWM controls the opening and closing of the spray valve. This process is repeated as the spray device moves to the next column after completing the spraying action of one column, thus completing the spraying of the entire board.

[0035] The sample loading control method and system for PCR reaction plate preparation proposed in this invention solves the problems of long preparation time, high cost, and easy impact on accuracy in the existing technology of PCR reaction plate preparation.

[0036] The present invention has the following advantages compared with the prior art: 1. Wide range of applications: Since the spray nozzle position of each spray needle can be edited, it can meet most current sample addition requirements without changing the hardware, and quickly realize the change of sample addition scheme.

[0037] 2. High scalability: Due to the scalability of this technology, devices with different numbers of injection needles can be developed based on this technology, and it can also be adapted to different reaction plate specifications.

[0038] 3. Reduced plate preparation time: Since each spray needle works independently during sample addition, the sample addition process can be completed in one batch to meet the sample addition requirements of a whole plate without the need for batch sample addition.

[0039] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the technical principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A method for controlling sample loading in PCR reaction plate preparation, characterized in that, include: Obtain the hole position information of each spray needle in the spray device on the reaction plate that needs to be sprayed. The spray device includes multiple spray needles, each of which corresponds to a sample type. Based on the orifice information and the preset movement speed, a motion control program for the current spray needle is generated; the motion control program is used to control the spray needle to pass through at least each orifice in the orifice information on the reaction plate at the preset movement speed. Based on the hole position information, calculate the hole spacing between two adjacent holes that need to be sprayed in the reaction plate that the current spray needle passes through in sequence. The PWM cycle for the liquid spraying operation of the spray needle is calculated based on the hole spacing, hole position information and preset movement speed. Based on the PWM cycle, a spray control program for the current spray needle is generated. The spray control program is used to control the current spray needle to add samples to the corresponding hole position according to the PWM cycle. Simultaneously execute motion control program and liquid spraying control program.

2. The sample loading control method for PCR reaction plate preparation according to claim 1, characterized in that, Before executing the motion control program and the liquid spray control program, the following is also included: Initialize the virtual orifice position, which is the first orifice position in each column on the reaction plate; or, the virtual orifice position is located outside the first orifice in each column on the reaction plate where liquid spraying begins, and is one orifice spacing away from the first orifice. The initial virtual orifice position is set as the starting point for the movement of the injection needle, as well as the reference point for timing the start of the PWM cycle and motion control.

3. The sample loading control method for PCR reaction plate preparation according to claim 1, characterized in that, The PWM cycle for the liquid spraying operation of the spray needle is calculated based on the hole spacing, hole position information, and preset movement speed, including: When only one hole spacing is calculated based on the hole position information, a PWM cycle is independently calculated based on the preset motion speed and the hole spacing, according to the preset motion relationship. When multiple hole spacings are calculated based on hole position information, different PWM cycles corresponding to each hole spacing are independently calculated based on a preset motion relationship, according to a preset motion speed and each different hole spacing. The preset motion relationship is as follows: Hole spacing = movement speed × PWM cycle.

4. The sample loading control method for PCR reaction plate preparation according to claim 3, characterized in that, This also includes optimizing the movement speed of the injection needle based on the orifice location information, including: Based on the orifice information, the movement trajectory of the spray needle on the reaction plate is divided into a continuous spray segment and a continuous non-spray segment, both of which include the case of a single orifice. The spray needle moves at a first speed in the continuous spray segment; The spray needle moves at a second speed in the continuous non-spraying section; The second speed is equal to or greater than the first speed.

5. The sample loading control method for PCR reaction plate preparation according to claim 1, characterized in that, The motion control program controls the spray needle to move from the first hole in the current column of the reaction plate that needs to be sprayed to the last hole in the current column that needs to be sprayed, and then from the last hole in the next column to the first hole in that column, and so on. The spray needle moves back and forth along the reaction plate column by column until it passes through every hole in the hole position information.

6. A sample loading control method for PCR reaction plate preparation according to claim 1 or 5, characterized in that, The motion control program is used to control the spray needle to pass through all the holes on the reaction plate at a preset speed.

7. The sample loading control method for PCR reaction plate preparation according to claim 1, characterized in that, Simultaneously execute motion control program and liquid spraying control program, including: Motion control commands are generated based on the motion control program; The drive motor used to drive the spray needle is controlled according to motion control commands; Generate spray control commands based on the spray control program; The spray valve of the spray needle is controlled according to the spray control command.

8. The sample loading control method for PCR reaction plate preparation according to claim 7, characterized in that, A row of spray needles is integrated into a single structure and driven by the same drive motor.

9. A system for a sample loading control method for PCR reaction plate preparation as described in any one of claims 1-8, characterized in that, include: The information acquisition module is used to acquire the hole position information of each spray needle in the spray device on the reaction plate that needs to be sprayed. The spray device includes multiple spray needles, each of which corresponds to a sample type. The first generation module is used to generate a motion control program for the current spray needle based on the orifice information and the preset motion speed; the motion control program is used to control the spray needle to pass through at least each orifice in the orifice information on the reaction plate at the preset motion speed. The first calculation module is used to calculate the hole spacing between two adjacent holes that need to be sprayed in the reaction plate through which the current spray needle passes, based on the hole position information. The second calculation module is used to calculate the PWM cycle of the liquid spraying operation of the liquid spraying needle based on the hole spacing, hole position information and preset movement speed. The second generation module is used to generate a liquid spraying control program for the current liquid spraying needle according to the PWM cycle. The liquid spraying control program is used to control the current liquid spraying needle to add samples to the corresponding hole position according to the PWM cycle. An execution control module is used to simultaneously execute a motion control program and a liquid spraying control program.

Images