TRACE METER AND METHOD FOR CALIBRATING DYNAMIC ULTRA-MICRO PIPETTING DEVICE
20250058312 ยท 2025-02-20
Assignee
Inventors
- Lihua Zhou (Chengdu, CN)
- Jiangwen TANG (Chengdu, CN)
- Chuandi Xu (Chengdu, CN)
- Zhanyue Jiang (Chengdu, CN)
Cpc classification
B01L2200/148
PERFORMING OPERATIONS; TRANSPORTING
B01L3/0213
PERFORMING OPERATIONS; TRANSPORTING
B01L2200/147
PERFORMING OPERATIONS; TRANSPORTING
International classification
Abstract
A trace meter and a method for calibrating a dynamic ultra-micro pipetting device are provided, which relate to the technical field of dynamic pipetting precision calibration. The trace meter includes a measuring compartment, a temperature sensor, a pressure sensor and a controller. A top of the measuring compartment is open, the temperature sensor is provided at a bottom of the measuring compartment, the pressure sensor is provided below the measuring compartment, both the temperature sensor and the pressure sensor are electrically connected with the controller, the temperature sensor is configured to measure the temperature of liquid in the measuring compartment, and the pressure sensor is configured to measure the weight of liquid in the measuring compartment.
Claims
1. A trace meter, comprising a measuring compartment (1), a temperature sensor (2), a pressure sensor (3) and a controller (4), wherein a top of the measuring compartment (1) is open, the temperature sensor (2) is provided at a bottom of the measuring compartment (1), the pressure sensor (3) is provided below the measuring compartment (1), both the temperature sensor (2) and the pressure sensor (3) are electrically connected with the controller (4), the temperature sensor (2) is configured to measure a temperature of liquid in the measuring compartment (1), and the pressure sensor (3) is configured to measure a weight of the liquid in the measuring compartment (1).
2. The trace meter according to claim 1, wherein an inner bottom wall of the measuring compartment (1) is provided with a lowest inclination point, the lowest inclination point is located directly below a center of gravity of the measuring compartment (1), a measuring hole (5) is formed at the lowest inclination point of the bottom wall of the measuring compartment (1), one side, adjacent to an inside of the measuring compartment (1), of the measuring hole (5) is covered with a flexible heat-conducting film (6), and a sensing end of the temperature sensor (2) is placed in the measuring hole (5) and abuts against a lower surface of the flexible heat-conducting film (6).
3. The trace meter according to claim 1, wherein a steam shield (7) covers above the measuring compartment (1), and a through hole (8) is formed in the steam shield (7) for a pipetting head (16) to pass through.
4. The trace meter according to claim 3, further comprising a bottom shell (9) provided below the measuring compartment (1), wherein the pressure sensor (3) is provided in the bottom shell (9), the bottom shell (9) is provided with a windshield (10), the windshield (10) is configured to cover the measuring compartment (1), a dripping hole (11) is formed in a top wall of the windshield (10) for the pipetting head (16) to pass through, a protective plate (12) is provided above the windshield (10), a passage hole (13) is formed in the protective plate (12), an aperture of the through hole (8) and an aperture of the dripping hole (11) are both larger than an aperture of the passage hole (13), a center line of the dripping hole (11), a center line of the through hole (8) and a center line of the measuring hole (5) are overlapped with each other, and gaps are left between the measuring compartment (1) and the windshield (10) and between the steam shield (7) and the windshield (10), respectively.
5. The trace meter according to claim 1, wherein a weighing plate (14) is provided below the measuring compartment (1), the pressure sensor (3) is located below the weighing plate (14), a positioning groove (15) is formed in the weighing plate (14), the positioning groove (15) comprises a concave conical surface and a horizontal inner bottom wall, an outer bottom wall of the measuring compartment (1) is provided with a positioning portion, a shape of the positioning portion is matched with a shape of the positioning groove (15), the positioning portion is located in the positioning groove (15), and the measuring hole (5) of the measuring compartment (1) is located directly above the horizontal inner bottom wall of the positioning groove (15).
6. A method for calibrating a dynamic ultra-micro pipetting device, wherein the method calibrates the dynamic ultra-micro pipetting device by using the trace meter according to claim 1, the method comprises following steps: S1, moving a pipetting head to a target position; S2, moving the trace meter to locate a dripping hole directly below the pipetting head; S3, placing standard liquid at a position where liquid is to be taken; S4, clearing measurement data of the trace meter to zero; S5, operating a same pipetting head to dynamically pipet liquid into a measuring compartment for n times, and reading a temperature data and a weight data of the liquid after each pipetting, wherein n is total pipetting times in a single channel; S6, obtaining n groups of measurement data of the dynamic pipetting process in the single channel, checking and correcting the measurement data, and calculating consistency of pipetting in the single channel by the controller; S7, removing the standard liquid in the measuring compartment; S8, repeating Step S1 to Step S7 to measure a next channel; S9, after measuring all channels, calculating a total standard deviation of dynamic high-throughput pipetting.
7. The method for calibrating the dynamic ultra-micro pipetting device according to claim 6, wherein in the Step S6, the controller calculates a liquid volume in the measuring compartment, wherein the liquid volume is
8. The method for calibrating the dynamic ultra-micro pipetting device according to claim 7, wherein in the Step S6, the controller corrects the liquid volume V.sub.x, wherein a volume correction value is V.sub.c=a+bV.sub.x, a measurement result of a corrected volume is V=Vc+V.sub.x, V is a final measurement result of the liquid volume after current pipetting, and a and b are correction coefficients of periodic calibration management and are preset in the controller after being measured and calibrated by a calibration unit at an upper level.
9. The method for calibrating the dynamic ultra-micro pipetting device according to claim 8, wherein in the Step S6, the consistency of pipetting in the single channel is
10. The method for calibrating the dynamic ultra-micro pipetting device according to claim 9, wherein in the Step S9, the total standard deviation of the dynamic high-throughput pipetting is
11. The method for calibrating the dynamic ultra-micro pipetting device according to claim 6, wherein an inner bottom wall of the measuring compartment (1) is provided with a lowest inclination point, the lowest inclination point is located directly below a center of gravity of the measuring compartment (1), a measuring hole (5) is formed at the lowest inclination point of the bottom wall of the measuring compartment (1), one side, adjacent to an inside of the measuring compartment (1), of the measuring hole (5) is covered with a flexible heat-conducting film (6), and a sensing end of the temperature sensor (2) is placed in the measuring hole (5) and abuts against a lower surface of the flexible heat-conducting film (6).
12. The method for calibrating the dynamic ultra-micro pipetting device according to claim 6, wherein a steam shield (7) covers above the measuring compartment (1), and a through hole (8) is formed in the steam shield (7) for a pipetting head (16) to pass through.
13. The method for calibrating the dynamic ultra-micro pipetting device according to claim 12, wherein the trace meter further comprises a bottom shell (9) provided below the measuring compartment (1), the pressure sensor (3) is provided in the bottom shell (9), the bottom shell (9) is provided with a windshield (10), the windshield (10) is configured to cover the measuring compartment (1), a dripping hole (11) is formed in a top wall of the windshield (10) for the pipetting head (16) to pass through, a protective plate (12) is provided above the windshield (10), a passage hole (13) is formed in the protective plate (12), an aperture of the through hole (8) and an aperture of the dripping hole (11) are both larger than an aperture of the passage hole (13), a center line of the dripping hole (11), a center line of the through hole (8) and a center line of the measuring hole (5) are overlapped with each other, and gaps are left between the measuring compartment (1) and the windshield (10) and between the steam shield (7) and the windshield (10), respectively.
14. The method for calibrating the dynamic ultra-micro pipetting device according to claim 6, wherein a weighing plate (14) is provided below the measuring compartment (1), the pressure sensor (3) is located below the weighing plate (14), a positioning groove (15) is formed in the weighing plate (14), the positioning groove (15) comprises a concave conical surface and a horizontal inner bottom wall, an outer bottom wall of the measuring compartment (1) is provided with a positioning portion, a shape of the positioning portion is matched with a shape of the positioning groove (15), the positioning portion is located in the positioning groove (15), and the measuring hole (5) of the measuring compartment (1) is located directly above the horizontal inner bottom wall of the positioning groove (15).
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0030] The drawings described herein are provided to provide a further understanding of the embodiments of the present disclosure and constitute a part of this application, and do not constitute limitations of the embodiments of the present disclosure.
[0031] In the figures:
[0032]
[0033]
[0034]
[0035]
[0036] Reference numerals in the drawings and names of corresponding parts: [0037] 1 measuring compartment; 2 temperature sensor; 3 pressure sensor; 4 controller; 5 measuring hole; 6 flexible heat-conducting film; 7 steam shield; 8 through hole; 9 bottom shell; 10 windshield; 11 dripping hole; 12 protective plate; 13 passage hole; 14 weighing plate; 15 positioning groove; 16 pipetting head; 17 upper computer; 18 analog-to-digital converter; 19 microcontroller unit; 20 external interface; 21 power module.
DETAILED DESCRIPTION OF THE EMBODIMENTS
[0038] In order to make the purpose, the technical solution and the advantages of the present disclosure more clear, the present disclosure will be further described in detail in conjunction with embodiments and attached drawings. The illustrative embodiments of the present disclosure and their descriptions are only used for explaining the present disclosure, and are not used as limitations of the present disclosure. It should be noted that the present disclosure has been in the actual development and use stage.
Embodiment 1
[0039] As shown in
[0040] As a preferred embodiment, as shown in
[0041] As a preferred embodiment, as shown in
[0042] As a preferred embodiment, as shown in
[0043] As a preferred embodiment, as shown in
[0044] As shown in
[0045] As shown in
[0046] As shown in
[0047] The embodiment further provides a method for calibrating a dynamic ultra-micro pipetting device by using the trace mete, as shown in
[0048] In S1, a pipetting head 16 is moved to a target position.
[0049] In S2, moving the trace meter is moved to locate a dropping hole 11 directly below the pipetting head 16.
[0050] In S3, standard liquid is placed at the position where liquid is to be taken, where the standard liquid adopts a pure water medium;
[0051] In S4, the temperature and weight data previously measured by the trace meter is cleared to zero.
[0052] In S5, the pipetting head 16 is operated to descend and extends into the measuring compartment, and liquid is dynamically pipetted into the measuring compartment 1 by the same pipetting head 16 for 10 times, and the temperature data and the weight data of the liquid are read after each pipetting by the microcontroller unit 19.
[0053] In S6, 10 groups of measurement data in a single channel transmitted by the microcontroller unit 19 is obtained, the 10 groups of measurement data in the single channel are checked and corrected, and the consistency of pipetting of the measurement data in the single channel is calculated by the controller 4;
[0054] In S7, standard liquid in the measuring compartment 1 is removed;
[0055] In S8, the Step S1 to the Step S7 are repeated to measure the next channel;
[0056] In S9, after measuring all channels, the total standard deviation of dynamic high-throughput pipetting is calculated by the controller 4.
[0057] In the Step S6 and the Step S9, the controller 4 checks, corrects and calculates the data, and all the data are uploaded to the upper computer for recording and storage.
[0058] Further, in the Step S6, the controller 4 checks the temperature data. The temperature
[0059] measurement value is calculated by a formula where t is the temperature measurement value, and to is the temperature measurement lower limit value of the temperature sensor 2 in the unit of C.; X.sub.t is the output value of the analog-to-digital converter (ADC) 18 at the current temperature; X.sub.t0 is the output value corresponding to the respective analog-to-digital converter at the temperature measurement lower limit value; R.sub.t is the resolution of the temperature measuring circuit consisted of the temperature sensor 2 and the analog-to-digital converter, that is, the corresponding change amount of the output data corresponding to the analog-to-digital converter every time there is a change of 1 C.
[0060] Further, in the Step S6, the controller 4 checks the weight data, and the weighing value is calculated by the formula
where mx is the mass measurement value, m.sub.0 is the weight measurement lower limit value of the pressure sensor 3 in the unit of g; X.sub.m is the output value of the analog-to-digital converter (ADC) 18 at the current temperature; X.sub.m0 is the output value corresponding to the respective analog-to-digital converter at the temperature measurement lower limit value; R.sub.m is the resolution of the weighing circuit consisted of the pressure sensor 3, the corresponding analog-to-digital converter and the weighing circuit, that is, the corresponding change amount of the output data corresponding to the analog-to-digital converter every time there is a change of 1 C.
[0061] Further, in the Step S6, the controller 4 checks the liquid volume V.sub.x in the measuring compartment 1, where the liquid volume is
V.sub.x is the liquid volume after each pipetting in the single channel, mx is the mass of the measured liquid, and t is the temperature of the measured liquid. The temperature value input by the operator or the temperature value obtained in Step S5 can be used.
[0062] Further, in the Step S6, the controller corrects the liquid volume V.sub.x, where the volume correction value is V.sub.c=a+bV.sub.x, the measurement result of the corrected volume is V=V.sub.c+V.sub.x, V is the final measurement result of the liquid volume after the current pipetting, and a and b are the correction coefficients of periodic calibration management and are preset in the controller 4 after the trace meter is measured and calibrated by a calibration unit at an upper level. In this embodiment, the linear fitting method is used to correct the volume in order to reduce the comprehensive error of the liquid volume measurement result of the trace meter. The conventional correction method mostly uses a broken line fitting method, which needs to input standard values and measurement values of all calibration points one by one. It is inconvenient to operate. The linear fitting method is used to correct the volume, which only needs to input two constants, which is simple to operate and stable in correction effect, and is convenient to improve the accuracy of the measurement data of the trace meter.
[0063] Further, in the Step S6, the controller 4 calculates the consistency of pipetting in the single channel. The consistency of pipetting in a single channel is
V.sub.i is the volume of the i-th pipetting in the current channel,
where i=1, 2, 3, . . . , n, n is the total times of pipetting of the current channel. When the single channel performs pipetting for 10 times, n=10.
[0064] Further, in the Step S9, the controller 4 calculates the total standard deviation of the dynamic high-throughput pipetting. The total standard deviation of dynamic high-throughput pipetting is
V.sub.ji is the volume of the i-th pipetting in a j-th channel, i=1, 2, 3, . . . n, n is the total times of pipetting of the single channel. When the single channel performs pipetting for 10 times, n=10. j=1, 2, 3, . . . m, m is the number of the measured channels, and
[0065] During each pipetting measurement reading, the system automatically makes ten consecutive readings and calculates the average value. For example, when pipetting in the first channel, the average value of ten consecutive readings is 9.9454 L9.945 L, and the volume reading data of each pipetting in the first channel is shown in the following table:
TABLE-US-00001 the volume of each pipetting (L) V.sub.1 V.sub.2 V.sub.3 V.sub.4 V.sub.5 V.sub.6 V.sub.7 V.sub.8 V.sub.9 V.sub.10 9.9455 9.9454 9.9455 9.9453 9.9453 9.9454 9.9453 9.9454 9.9453 9.9454
[0066] According to the formula
the average value of the volume of each pipetting is substituted into V in the above formula, and the volume data of each pipetting is substituted into V.sub.i in the above formula. The uncertainty of Class A of the measurement result is calculated as U.sub.A=0.08 nL, which is rounded to U.sub.A=0.10 nL.
[0067] A four-channel dynamic pipetting device is calibrated according to the method of this embodiment, and according to the Step S1 to the Step S8, the measured original data is obtained as follows, where V.sub.i is the volume of each pipetting of the current channel,
and i=1, 2, 3, . . . n. V is the final measurement result of the total liquid volume after the current pipetting.
[0068] When calibrating the four channels, the volume of each pipetting is shown in the following table:
TABLE-US-00002 the nominal value of the volume of the volume of each pipetting (L) pipetting channel V.sub.1 V.sub.2 V.sub.3 V.sub.4 V.sub.5 V.sub.6 V.sub.7 V.sub.8 V.sub.9 V.sub.10 10 (L) 1 9.945 9.936 9.912 9.923 9.924 9.936 9.919 9.945 9.928 9.918 2 9.936 9.919 9.936 9.946 9.955 9.928 9.924 9.956 9.937 9.938 3 9.953 9.960 9.946 9.952 9.935 9.963 9.954 9.960 9.934 9.952 4 9.942 9.958 9.959 9.961 9.932 9.946 9.937 9.948 9.959 9.961
[0069] According to the formula
the consistency of pipetting in the single channel is calculated, as shown in the following table:
TABLE-US-00003 the nominal absolute relative value of the pipetting pipetting volume of consistency consistency pipetting (L) channel (L) (%) 10 1 0.0115 0.11 2 0.0122 0.12 3 0.0099 0.10 4 0.0108 0.11
[0070] According to the formula
the absolute total standard deviation of the high-throughput pipetting is calculated as 0.005 L, and the relative total standard deviation relative to the nominal value of the volume of pipetting is 0.5%.
[0071] According to the formula
the average value of the volume of high-throughput pipetting is calculated as
[0072] According to the indication error=measurement indication value-reference value, the indication error of the high-throughput pipetting device measured this time is calculated as E.sub.a=0.058 L, and the correction value is C=0.058 L at the working point of 10 L.
[0073] According to the indication range=maximum positive deviation+maximum negative deviation, the range corrected in this measurement is calculated as R=0.051 L.
[0074] It can be seen that the present disclosure realizes the measurement of precision-related indexes such as the consistency y of pipetting in a single channel, the total standard deviation , the indication error E.sub.a, and the range R of the trace-level ultra-high-precision and high-throughput pipetting device, and can give the correction value C to calibrate the dynamic pipetting device. The uncertainty of Class A of the measurement result can be up to 1 nL, which is more than one order of magnitude higher than that of the conventional measurement method.
[0075] In the solution, according to the method for calibrating the dynamic ultra-micro pipetting device by using the trace meter, the consistency measurement of pipetting in the single channel and the total standard deviation measurement of the high-throughput pipetting device are achieved, such that a high-precision value assurance technology is provided to the high-throughput pipetting device and the pipetting operation, which is beneficial to popularizing the high-throughput metering technology in the high-precision application field. The trace-level ultra-high precision and high-throughput pipetting precision measurement is achieved, in which the volume resolution is up to 0.1 nL and the measurement uncertainty is better than 1 nL which is more than one order of magnitude higher than that of the conventional measurement method. The meaning of the uncertainty refers to the degree that the measured value cannot be affirmed due to the existence of the measurement error. Conversely, the uncertainty also shows the reliability of the result, which is an index of the quality of the measurement result. The smaller the uncertainty, the higher the quality, the higher the level, and the higher its use value. The greater the uncertainty, the lower the quality of the measurement result, the lower the level, and the lower its use value. When reporting the measurement result of physical quantities, the corresponding uncertainty must be given, which is convenient for users to evaluate their reliability on the one hand and enhances the comparability of the measurement results on the other hand. The automatic calibration of pipetting precision of the pipetting device is achieved, which greatly improves the calibration efficiency of the pipetting device.
[0076] The above-mentioned specific embodiments further explain the purpose, the technical solution and the beneficial effect of the present disclosure in detail. It should be understood that the above-mentioned embodiments are only specific embodiments of the present disclosure and are not used to limit the scope of protection of the present disclosure. Any modification, equivalent substitution, improvement, etc. made within the spirit and principle of the present disclosure should be included in the scope of protection of the present disclosure.