A method for calibrating a flow cytometer detector

Abstract

The present invention provides a method for calibrating a flow cytometer detector, which belongs to the technical field of instrument calibration, and includes: installing the light source, filter, and light source sensor of the flow cytometer at corresponding positions through positioning pin holes, and after fixing, connecting a motor to an eccentric wheel, and at this time, the motor connected to the eccentric wheel is recorded as zero position; connecting a sensor panel, and the sensor panel receives the outgoing light source in the filter, and calculates the position error d between the actual light source and the theoretical light source according to the coordinate positioning of the sensing area of ​​the sensor panel; feeding back the position error d to a control system, and determining the deflection angle of the filter and the adjustment angle of the eccentric wheel, and combining with an error setting function, controlling the motor to drive the eccentric wheel to correct the filter; after calibration, checking the position deviation, if the deviation threshold is met, stopping the calibration, if not, continuing the calibration until the deviation threshold is met, and the calibration ends. Automatic calibration is realized to improve the calibration efficiency and ensure the detection accuracy.

Description

Technical Field

[0001] The invention relates to the technical field of instrument calibration, and in particular to a method for calibrating a flow cytometer detector. Background Art

[0002] The flow cytometer detector collects the spectrum of the required frequency through the filter. The light source can pass through the filter of the corresponding bandwidth. The back-end APD module will convert the collected corresponding light signal into an electrical signal to obtain the information in the cell. Otherwise, it will be reflected to the next filter and decompose the information of each bandwidth of the light source in turn. Currently, most detectors on the market use mechanical structures to ensure the accuracy of the optical path. Manual calibration of the optical path and detector accuracy is time-consuming and laborious.

[0003] Therefore, the present invention provides a method for calibrating a flow cytometer detector. Summary of the invention

[0004] The present invention provides a method for calibrating a flow cytometer detector, which is used to determine the deflection angle and the adjustment angle by automatically calculating the existing position error, and further combines the error setting function to control the motor to drive the eccentric wheel to automatically correct, thereby realizing automatic calibration to improve the calibration efficiency, and performing continuous calibration by judging with the deviation threshold to ensure the detection accuracy.

[0005] The present invention provides a method for calibrating a flow cytometer detector, comprising:

[0006] Step 1: Install the light source, filter, and light source sensor of the flow cytometer at the corresponding positions through the positioning pin holes, wherein the filter is initially roughly positioned through the eccentric wheel and the rotation center. After being fixed, the motor is connected to the eccentric wheel. At this time, the motor connected to the eccentric wheel is recorded as zero position;

[0007] Step 2: Turn on the sensor panel, the sensor panel receives the outgoing light source in the filter, locates the coordinates of the sensing area of ​​the sensor panel, and calculates the position error d between the actual light source and the theoretical light source;

[0008] Step 3: Feedback the position error d to the control system, determine the deflection angle of the filter and the adjustment angle of the eccentric wheel, and combine the error setting function to control the motor to drive the eccentric wheel to correct the filter;

[0009] Step 4: After calibration, check the position deviation. If the deviation threshold is met, stop calibration. If not, continue calibration until the deviation threshold is met and calibration ends.

[0010] Preferably, calculating the position error d between the actual light source and the theoretical light source according to the coordinate positioning of the sensing area of ​​the sensor panel includes:

[0011] Get the theoretical coordinates based on the sensing area ;

[0012] Capture actual coordinates based on the sensing area ;

[0013] Depends on the theoretical coordinates With actual coordinates , and obtain the position error d.

[0014] Preferably, determining the deflection angle of the filter and the adjustment angle of the eccentric wheel includes:

[0015] According to a first relationship expression between the optical filter and the position error d, a deflection angle of the optical filter is obtained;

[0016] According to the second relationship expression between the optical filter and the eccentric wheel, the adjustment angle of the eccentric wheel is obtained.

[0017] Preferably, the method further comprises: establishing an error setting function, comprising:

[0018] Capturing the spot position and spot shape of the outgoing light source in the sensing area of ​​the sensor panel, and constructing a spot matrix based on the sensing area;

[0019] The point of the theoretical coordinate of the sensing area is locked, and the light spot matrix is ​​analyzed in the up, down, left, right, upper left, lower left, upper right, and lower right directions, and the light spot occupied area and the light spot contour line length in each direction are obtained respectively to obtain the first deviation coefficient of the corresponding direction;

[0020] ;

[0021] in, represents the first deviation coefficient in the i1th direction; Represents the theoretical spot area in the i1th direction; represents the area occupied by the light spot in the i1th direction; Indicates the length of the light spot contour line in the i1th direction; Indicates that the i1th direction is The theoretical length determined for the radius; is a constant, with a value of 3.14; Indicates the preset offset area in the i1th direction; ln indicates the sign of the logarithmic function; the value of i is 1, 2, 3, 4, 5, 6, 7, 8;

[0022] All first deviation coefficients are divided according to the deviation level division table, and an initial analysis vector is constructed according to the number of occurrences of the first deviation coefficients involved in the same division level;

[0023] Inputting the initial analysis vector into a deviation analysis model to obtain a reference error;

[0024] According to the current setting combination of all filters of the flow cytometer and in combination with the light direction of the incident light source, a first analysis vector is constructed, and input into a forward analysis model to obtain first setting deviation information of each filter;

[0025] According to the current setting combination of all filters of the flow cytometer and in combination with the light direction of the outgoing light source, a second analysis vector is constructed, and input into the reverse analysis model to obtain the second setting deviation information of each filter;

[0026] Based on the first setting deviation information and the second setting deviation analysis, sequentially determining the self-light source error of each filter;

[0027] According to the own light source error of each filter and combined with the reference error, an error setting function is constructed.

[0028] Preferably, determining the self-light source error of each filter in turn includes:

[0029] Extracting the first deviation of the corresponding filter from the first setting deviation information and the second setting deviation information respectively and the second deviation ;

[0030] when When , the error of the own light source is determined to be 0;

[0031] when When , the error of the own light source is determined to be r1;

[0032] when ,according to Lock the minimum value, when the minimum value is When It is determined to be the self-light source error, where m0 represents the total number of filters; Indicates that all filters are based on the first deviation and the second deviation The variance of the absolute value of the difference; Indicates that all filters are based on the first deviation The variance of Indicates that all filters are based on the second deviation The variance of , and r1, r2 are variables; min represents the minimum value symbol;

[0033] When the minimum value is When Determined to be self-light source error;

[0034] When the minimum value is When It is determined to be the error of the own light source, where Indicates the first deviation of the jth filter and the second deviation The absolute value of the difference;

[0035] when When the error is not zero, the error of the light source itself is determined.

[0036] Preferably, controlling the motor to drive the eccentric wheel to correct the filter comprises:

[0037] Performing a first matching operation on the error setting function and the function-filter comparison table to obtain a first calibration solution, and performing a second matching operation on the error setting function and the function-eccentric wheel comparison table to obtain a second calibration solution;

[0038] Based on the first calibration scheme and the second calibration scheme, respectively optimizing the deflection angle of the filter and the adjustment angle of the eccentric wheel;

[0039] Based on the optimization result, the motor is controlled to drive the eccentric wheel to correct the filter.

[0040] Preferably, based on the first calibration scheme and the second calibration scheme, the deflection angle of the filter and the adjustment angle of the eccentric wheel are optimized respectively, including:

[0041] Extracting a first error angle for the optical filter from the first calibration scheme, and obtaining a second error angle based on the eccentric wheel according to a second relationship expression between the optical filter and the eccentric wheel;

[0042] Extracting a third error angle for the eccentric wheel from the second calibration scheme, and obtaining a fourth error angle based on the filter according to a second relationship expression between the filter and the eccentric wheel;

[0043] Taking the average value of the first error angle and the fourth error angle as the first optimized angle of the filter;

[0044] Determining a second optimized angle for the eccentric wheel according to the first optimized angle, the second error angle, and the third error angle;

[0045] Recording the sum of the adjustment angle and the second optimization angle, the third optimization angle converted from the first optimization angle based on the second relationship expression, and the adjustment angle, respectively, to control the first position point and the second position point reached by the eccentric wheel after the movement ends;

[0046] Determine a first distance between the first position point and the theoretical coordinates, and at the same time, determine a second distance between the second position point and the theoretical coordinates;

[0047] According to min (first distance, second distance), the corresponding final optimization angle is selected as an optimization reference.

[0048] Preferably, the method further comprises: setting a deviation threshold, including:

[0049] Capture the moving path of the center point of the light spot on the sensor panel during the process of the control motor driving the eccentric wheel to correct the filter, divide the moving path equally to obtain several moving points, and determine the relationship between each moving point and the theoretical coordinates respectively Straight-line distance;

[0050] Determine the absolute value of the distance difference between the two adjacent moving points according to the order of the moving points, and average all the absolute values ​​of the distance differences to obtain the unit difference. ;

[0051] Obtaining the instrument attributes of the flow cytometer, and obtaining the initial threshold value yc0 from the attribute-threshold comparison table;

[0052] Determine the detection accuracy Jd according to the experimental purpose of the cells to be detected;

[0053] Based on the initial threshold yc0, the detection accuracy Jd and the unit difference , the absolute value of the distance difference between the last moving point and the second last moving point Ld, and the deviation threshold Py is obtained;

[0054] ;

[0055] in, represents the precision conversion function of the initial threshold yc0; n4 represents the number of moving points; Indicates the conversion factor between accuracy and threshold.

[0056] Compared with the prior art, the present invention has the following beneficial effects:

[0057] The deflection angle and adjustment angle are determined by automatically calculating the existing position error, and further combined with the error setting function, the motor is controlled to drive the eccentric wheel to automatically correct, so as to realize automatic calibration to improve the calibration efficiency, and continuous calibration is carried out by judging the deviation threshold to ensure the detection accuracy.

[0058] Other features and advantages of the present invention will be described in the following description, and partly become apparent from the description, or understood by practicing the present invention. The purpose and other advantages of the present invention can be realized and obtained by the structures particularly pointed out in the written description and the accompanying drawings.

[0059] The technical solution of the present invention is further described in detail below through the accompanying drawings and embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0060] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation of the present invention. In the accompanying drawings:

[0061] Figure 1 A flow chart of a method for calibrating a flow cytometer detector according to an embodiment of the present invention;

[0062] Figure 2 A structural diagram of the optical path of the detector in an embodiment of the present invention;

[0063] Figure 3 This is a schematic diagram of the optical path adjustment principle in an embodiment of the present invention;

[0064] Figure 4 The figure is a rotation structure diagram of the eccentric wheel in the embodiment of the present invention;

[0065] Figure 5 A geometric relationship diagram in an embodiment of the present invention;

[0066] Figure 6 A diagram showing the structure of the sensor and the motor in an embodiment of the present invention;

[0067] Figure 7 2 is a front and rear position diagram of an optical filter according to an embodiment of the present invention;

[0068] Figure 8 is a state diagram of different positions of the light spot in an embodiment of the present invention;

[0069] Fig. 9 4 is a structural diagram of a spectral detector filter calibration system in an embodiment of the present invention. DETAILED DESCRIPTION

[0070] The preferred embodiments of the present invention are described below in conjunction with the accompanying drawings. It should be understood that the preferred embodiments described herein are only used to illustrate and explain the present invention, and are not used to limit the present invention.

[0071] The present invention provides a method for calibrating a flow cytometer detector, such as Figure 1 As shown, including:

[0072] Step 1: Install the light source, filter, and light source sensor of the flow cytometer at the corresponding positions through the positioning pin holes, wherein the filter is initially roughly positioned through the eccentric wheel and the rotation center. After being fixed, the motor is connected to the eccentric wheel. At this time, the motor connected to the eccentric wheel is recorded as zero position;

[0073] Step 2: Turn on the sensor panel, the sensor panel receives the outgoing light source in the filter, locates the coordinates of the sensing area of ​​the sensor panel, and calculates the position error d between the actual light source and the theoretical light source;

[0074] Step 3: Feedback the position error d to the control system, determine the deflection angle of the filter and the adjustment angle of the eccentric wheel, and control the motor to drive the eccentric wheel to correct the filter in combination with the error setting function;

[0075] Step 4: After calibration, check the position deviation. If the deviation threshold is met, stop calibration. If not, continue calibration until the deviation threshold is met and calibration ends.

[0076] Preferably, calculating the position error d between the actual light source and the theoretical light source according to the coordinate positioning of the sensing area of ​​the sensor panel includes:

[0077] Get the theoretical coordinates based on the sensing area ;

[0078] Capture actual coordinates based on the sensing area ;

[0079] Depends on the theoretical coordinates With actual coordinates , and obtain the position error d.

[0080] Preferably, determining the deflection angle of the filter and the adjustment angle of the eccentric wheel includes:

[0081] According to a first relationship expression between the optical filter and the position error d, a deflection angle of the optical filter is obtained;

[0082] According to the second relationship expression between the optical filter and the eccentric wheel, the adjustment angle of the eccentric wheel is obtained.

[0083] In this embodiment, if Figure 2 As shown, 101 is a filter holder, 301-305, 401-405 are filters, 501 is incident light, 502 is outgoing light, and the straight line in the figure is the light path in the detector.

[0084] The specific adjustment principle is as follows Figure 3 As shown, 102 is the rotation center of the filter holder, and the filter holder 101 rotates around the filter holder rotation center 102. 1011 and 1012 are the two surfaces of the filter fixing the filter. 201 is a light sensor (CMOS), which can sense the position and shape of the light spot and transmit the data to the control system. Filters 301~305 are attached to surface 1011, and filters 401~405 are attached to surface 1012. The incident light 501 is irradiated onto the filter 301 through the filter holder rotation center 102, and the outgoing light 502 is emitted through the hole on the surface 1012 and irradiates the surface of the sensor 201. The red border is the initial state of the filter. The coordinate system is established with the filter holder rotation center 102 as the coordinate origin. The X and Y axes are as shown in Figure 3 As shown. In the coordinate system, L1 represents the incident light 501, the incident angle is α, which is fixed; L2 represents the initial state of the outgoing light 502; L3 represents the surface 1011, and θ is the rotation angle of the filter around the filter holder rotation center 102; when the filter rotates, the outgoing light 502 changes from L2 to L4, and the X-direction displacement irradiated on the sensor 201 is recorded as d; the Y-direction distance between the surface 1011 and the surface 1012 is 2a, and the initial state distance between the sensor and the surface 1011 is H.

[0085] Knowing the above variables and their meanings, we can get the rotation angle θ and the spot offset distance The relationship can be The offset of the rotation angle θ is known, and the information is transmitted to the control system, and the filter is calibrated by controlling the corresponding rotation angle.

[0086] The calculation process of the relationship between the rotation angle θ and the offset d is as follows: , where right is positive, left is negative, counterclockwise is positive, and clockwise is negative.

[0087] Incident light,

[0088] Initial emitted light,

[0089] Deflect the outgoing light,

[0090] Surface 1011, small range deflection can be approximated

[0091] L3, L4 focus is P1, then P1

[0092] From the geometric relationship, we can know that in the right triangle P1P2P3, we can approximately consider that: ,

[0093] The incident light remains unchanged, and the filter rotates by θ (counterclockwise is positive, clockwise is negative) around the filter holder rotation center 102. Then the angle change of the light path in the filter is 2θ, that is, the angle is , , , we can know the relationship expression between the deflection angle θ and the spot displacement d (the first relationship expression).

[0094] .

[0095] like Figure 4As shown, the base u03 has two pin holes, and the eccentric wheel u01 is installed on the base u03 through the pin holes and can rotate relatively. One end of the filter u02 cooperates with the rotation center 102 of the filter bracket, and the other end cooperates with the eccentric wheel u01. By rotating the eccentric wheel u01, the filter u02 can be rotated to a certain angle accordingly. The dotted line figure in the figure is the initial state, and the realized figure is the state after the eccentric wheel u01 is rotated.

[0096] like Figure 5 As shown, the distance between the rotation center of the eccentric wheel u01 and the rotation center of the filter holder 102 is R, the eccentric distance is e, the rotation angle of the eccentric wheel is η, and the corresponding rotation angle of the filter u02 is θ. According to the geometric relationship, the relationship between θ and η (the second relationship expression) can be obtained:

[0097] .

[0098] From the above two expressions, it is known that the spot offset d can be obtained by The deflection angle of filter u02 is θ, It can be seen that the eccentric wheel u01 needs to rotate an angle to correct the optical path.

[0099] like Figure 6 As shown, the optical sensor 201 is fixed at the theoretical position, the motor 601 is connected to the eccentric wheel u01, and the center of the sensitive area of ​​the optical sensor 201 is its own coordinate origin. The displacement of the light spot position can be determined, and the position change information can be transmitted to the system. The system controls the motor to rotate the corresponding angle to calibrate the position of the filter u02. Figure 7 As shown, the v1 and v2 frames represent the filter positions in the previous and next states, respectively. Figure 8 As shown, the grayscale value represents the state of the light spot at different positions in the sensor area.

[0100] Spectral detector filter calibration system such as Fig. 9 As shown, light source 701 is mounted and fixed on bracket 801, sensor 201 is installed at a specified position through positioning pins, the positioning holes of fixed bracket 801 and sensor 201 can be processed to ensure relative position, filter u02 is initially positioned through positioning pins and eccentric wheel u01, and the filter calibration can be completed by connecting eccentric wheel u01 and rotating motor.

[0101] In this embodiment, the error setting function is determined based on the error of the filter itself.

[0102] The beneficial effects of the above technical solution are: by automatically calculating the existing position error, the deflection angle and the adjustment angle are determined, and further combined with the error setting function, the motor is controlled to drive the eccentric wheel to automatically correct, so as to realize automatic calibration to improve the calibration efficiency, and continuous calibration is performed through judgment with the deviation threshold to ensure the detection accuracy.

[0103] The present invention provides a method for calibrating a flow cytometer detector, further comprising: establishing an error setting function, including:

[0104] Capturing the spot position and spot shape of the outgoing light source in the sensing area of ​​the sensor panel, and constructing a spot matrix based on the sensing area;

[0105] The point of the theoretical coordinate of the sensing area is locked, and the light spot matrix is ​​analyzed in the up, down, left, right, upper left, lower left, upper right, and lower right directions, and the light spot occupied area and the light spot contour line length in each direction are obtained respectively to obtain the first deviation coefficient of the corresponding direction;

[0106]

[0107] in, represents the first deviation coefficient in the i1th direction; Represents the theoretical spot area in the i1th direction; represents the area occupied by the light spot in the i1th direction; Indicates the length of the light spot contour line in the i1th direction; Indicates that the i1th direction is The theoretical length determined for the radius; is a constant, with a value of 3.14; Indicates the preset offset area in the i1th direction; ln indicates the sign of the logarithmic function; the value of i is 1, 2, 3, 4, 5, 6, 7, 8;

[0108] All first deviation coefficients are divided according to the deviation level division table, and an initial analysis vector is constructed according to the number of occurrences of the first deviation coefficients involved in the same division level;

[0109] Inputting the initial analysis vector into a deviation analysis model to obtain a reference error;

[0110] According to the current setting combination of all filters of the flow cytometer and in combination with the light direction of the incident light source, a first analysis vector is constructed, and input into a forward analysis model to obtain first setting deviation information of each filter;

[0111] According to the current setting combination of all filters of the flow cytometer and in combination with the light direction of the outgoing light source, a second analysis vector is constructed, and input into the reverse analysis model to obtain the second setting deviation information of each filter;

[0112] Based on the first setting deviation information and the second setting deviation analysis, sequentially determining the self-light source error of each filter;

[0113] According to the own light source error of each filter and combined with the reference error, an error setting function is constructed.

[0114] In this embodiment, the sensing area can be a circular area, a pre-planned area obtained with the center point of the sensor panel as the origin, and the spot position refers to the actual position on the sensor panel, and the spot shape refers to the irregular shape or regular shape of the outer contour of the spot.

[0115] In this embodiment, the light spot matrix is ​​constructed in sequence according to the positions, with each position point in the sensing area as an element, and the row vector of the matrix is ​​the light spot value at the coordinates of each horizontal position point at the same vertical value. The light spot value is 0 or 1. When a light spot exists at the position point, it is regarded as 1, and when it does not exist, it is regarded as 0.

[0116] In this embodiment, when analyzing the up, down, left, right, upper left, lower left, upper right, and lower right directions, the sensing area is divided into 8 blocks according to the 8 directions, among which up, down, left, and right are all in the positive direction, and 360 degrees are divided into 45 degrees.

[0117] In this embodiment, the area occupied by the light spot and the length of the light spot contour line can be directly obtained according to the light spot matrix, that is, the area and line length of the region whose value is 1.

[0118] In this embodiment, the deviation level classification table includes different coefficient ranges and levels for the coefficient ranges, which are pre-set.

[0119] Initial analysis vector = {the first deviation coefficient and the number of occurrences at different division levels}.

[0120] In this embodiment, the deviation analysis model is obtained by training the neural network model based on different initial analysis vectors and analytical error results of the vectors as samples, so the reference error can be directly obtained.

[0121] In this embodiment, the current setting combination refers to the setting of the filter. Since the system error caused by the device itself cannot be avoided, the accuracy of subsequent control according to the angle is further improved by performing deviation analysis on the filter itself.

[0122] In this embodiment, the first analysis vector is obtained from the incident light source-combination-vector comparison table;

[0123] The second analysis vector is obtained from the outgoing light source-combination-vector comparison table;

[0124] It should be noted that the comparison table contains different light sources, combinations and setting vectors under the two, which are all pre-set.

[0125] In this embodiment, the forward analysis model is obtained by training the neural network model by forward analyzing the first analysis vector based on the incident light source to determine the error existing in the filter as a sample.

[0126] The reverse analysis model is obtained by reversely analyzing the second analysis vector based on the reflected light source to determine the error existing in the filter as a sample for training the neural network model.

[0127] Then the corresponding deviation information can be obtained, that is, the error information obtained by the model output.

[0128] In this embodiment, the error setting function=Ws(self-light source error of each filter, reference error).

[0129] The beneficial effects of the above technical solution are: on the one hand, the first deviation coefficient is determined by analyzing the light spot matrix in 8 directions, and then the reference error is effectively obtained through grade division; on the other hand, the self-light source error of each filter is determined by cutting in from the incident light source and the outgoing light source, so as to comprehensively construct the error setting function, which provides a basis for subsequent optimization control.

[0130] The present invention provides a method for calibrating a flow cytometer detector, which sequentially determines the self-light source error of each filter, comprising:

[0131] Extracting the first deviation of the corresponding filter from the first setting deviation information and the second setting deviation information respectively and the second deviation ;

[0132] when When , the error of the own light source is determined to be 0;

[0133] when When , the error of the own light source is determined to be r1;

[0134] when ,according to Lock the minimum value, when the minimum value is When It is determined to be the self-light source error, where m0 represents the total number of filters; Indicates that all filters are based on the first deviation and the second deviation The variance of the absolute value of the difference; Indicates that all filters are based on the first deviation The variance of Indicates that all filters are based on the second deviation The variance of , and r1, r2 are variables; min represents the minimum value symbol;

[0135] When the minimum value is When Determined to be self-light source error;

[0136] When the minimum value is When It is determined to be the error of the own light source, where Indicates the first deviation of the jth filter and the second deviation The absolute value of the difference;

[0137] when When the error is not zero, the error of the light source itself is determined.

[0138] In this embodiment, the deviation information is set to be the deviation amount.

[0139] The beneficial effect of the above technical solution is: by comparing and analyzing the first deviation and the second deviation, the self-light source errors in different situations are classified and discussed, providing a basis for constructing a position error function.

[0140] The present invention provides a method for calibrating a flow cytometer detector, which controls a motor to drive an eccentric wheel to calibrate a filter, comprising:

[0141] Performing a first matching operation on the error setting function and the function-filter comparison table to obtain a first calibration solution, and performing a second matching operation on the error setting function and the function-eccentric wheel comparison table to obtain a second calibration solution;

[0142] Based on the first calibration scheme and the second calibration scheme, respectively optimizing the deflection angle of the filter and the adjustment angle of the eccentric wheel;

[0143] Based on the optimization result, the motor is controlled to drive the eccentric wheel to correct the filter.

[0144] In this embodiment, the function-filter comparison table includes different error setting functions and calibration schemes of filters matching the functions, and the first calibration scheme includes a calibration error angle for the filter.

[0145] The function-eccentric wheel comparison table includes different error setting functions and calibration schemes of the eccentric wheel matching the functions, and the second calibration scheme includes the calibration error angle of the eccentric wheel.

[0146] The beneficial effect of the above technical solution is: calibration solutions based on the filter and the eccentric wheel are obtained from two comparison tables respectively to comprehensively ensure the optimization result, thereby improving the control accuracy.

[0147] The present invention provides a method for calibrating a flow cytometer detector, which optimizes the deflection angle of the filter and the adjustment angle of the eccentric wheel based on the first calibration scheme and the second calibration scheme, respectively, including:

[0148] Extracting a first error angle for the optical filter from the first calibration scheme, and obtaining a second error angle based on the eccentric wheel according to a second relationship expression between the optical filter and the eccentric wheel;

[0149] Extracting a third error angle for the eccentric wheel from the second calibration scheme, and obtaining a fourth error angle based on the filter according to a second relationship expression between the filter and the eccentric wheel;

[0150] Taking the average value of the first error angle and the fourth error angle as the first optimized angle of the filter;

[0151] Determining a second optimized angle for the eccentric wheel according to the first optimized angle, the second error angle, and the third error angle;

[0152] Recording the sum of the adjustment angle and the second optimization angle, the third optimization angle converted from the first optimization angle based on the second relationship expression, and the adjustment angle, respectively, to control the first position point and the second position point reached by the eccentric wheel after the movement ends;

[0153] Determine a first distance between the first position point and the theoretical coordinates, and at the same time, determine a second distance between the second position point and the theoretical coordinates;

[0154] According to min (first distance, second distance), the corresponding final optimization angle is selected as an optimization reference.

[0155] In this embodiment, the first error angle is the extracted calibration error angle of the optical filter.

[0156] The third error angle is the extracted calibration error angle of the eccentric wheel.

[0157] Since the first relational expression and the second relational expression exist, the corresponding second error angle and fourth error angle can be directly obtained.

[0158] In this embodiment, the first optimization angle=(first error angle+fourth error angle) / 2.

[0159] In this embodiment, the second optimization angle=(first optimization angle+(second error angle+third error angle) / 2) / 2.

[0160] In this embodiment, the first distance and the second distance are calculated based on the coordinates of two points, which belongs to the prior art.

[0161] In this embodiment, the first distance corresponds to the first position point, and the first position point corresponds to the sum of the adjustment angle and the second optimization angle;

[0162] The second distance corresponds to the second position point, and the second position point corresponds to the sum of the third optimization angle after converting the first optimization angle based on the second relational expression and the adjustment angle.

[0163] The beneficial effect of the above technical solution is: by obtaining the error angles of the filter and the eccentric wheel, the optimized angle for the eccentric wheel is obtained, that is, the two obtained optimized angles are adjusted to obtain the final optimized angle corresponding to the minimum distance, and optimized control is performed to ensure control accuracy.

[0164] The present invention provides a method for calibrating a flow cytometer detector, further comprising: setting a deviation threshold, including:

[0165] Capture the moving path of the center point of the light spot on the sensor panel during the process of the control motor driving the eccentric wheel to correct the filter, divide the moving path equally to obtain several moving points, and determine the relationship between each moving point and the theoretical coordinates respectively Straight-line distance;

[0166] Determine the absolute value of the distance difference between the two adjacent moving points according to the order of the moving points, and average all the absolute values ​​of the distance differences to obtain the unit difference. ;

[0167] Obtaining the instrument attributes of the flow cytometer, and obtaining the initial threshold value yc0 from the attribute-threshold comparison table;

[0168] Determine the detection accuracy Jd according to the experimental purpose of the cells to be detected;

[0169] Based on the initial threshold yc0, the detection accuracy Jd and the unit difference , the absolute value of the distance difference between the last moving point and the second last moving point Ld, and the deviation threshold Py is obtained;

[0170] ;

[0171] in, represents the precision conversion function of the initial threshold yc0; n4 represents the number of moving points; Indicates the conversion factor between accuracy and threshold.

[0172] It should be noted that .

[0173] In this embodiment, the moving route refers to the process route in which the motor drives the eccentric wheel to be corrected according to the final angle.

[0174] In this embodiment, the equal division may be to split the moving route into n4-1 points.

[0175] In this embodiment, the attribute-threshold comparison table includes different instrument attributes and initial thresholds matching the attributes, which are preset, and the initial threshold is for distance.

[0176] In this embodiment, different detection accuracies correspond to different experimental purposes, which is also pre-planned.

[0177] In this embodiment, for example, the attributes are immunophenotyping analysis, apoptosis analysis, and cytokine analysis of peripheral blood cells.

[0178] In this embodiment, the conversion coefficient is used to convert the accuracy to the threshold value to improve the accuracy of the deviation threshold calculation.

[0179] The beneficial effect of the above technical solution is: by equally dividing the moving route, the absolute value of the distance difference between adjacent moving points is determined, and then the unit difference is obtained, and the initial threshold under the attribute and the detection accuracy under the experimental purpose are combined to comprehensively obtain the deviation threshold, which improves the judgment basis for whether to continue correction in the future and indirectly improves the adjustment efficiency.

[0180] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A method for calibrating a flow cytometer detector, characterized in that: include: Step 1: Install the light source, filter, and light source sensor of the flow cytometer at the corresponding positions through the positioning pin holes, wherein the filter is initially roughly positioned through the eccentric wheel and the rotation center. After being fixed, the motor is connected to the eccentric wheel. At this time, the motor connected to the eccentric wheel is recorded as zero position; Step 2: Turn on the sensor panel, the sensor panel receives the outgoing light source in the filter, locates the coordinates of the sensing area of ​​the sensor panel, and calculates the position error d between the actual light source and the theoretical light source; Step 3: Feedback the position error d to the control system, determine the deflection angle of the filter and the adjustment angle of the eccentric wheel, and combine the error setting function to control the motor to drive the eccentric wheel to correct the filter; Step 4: After calibration, check the position deviation. If the deviation threshold is met, stop calibration. If not, continue calibration until the deviation threshold is met and calibration ends. The method further includes: establishing an error setting function, including: Capturing the spot position and spot shape of the outgoing light source in the sensing area of ​​the sensor panel, and constructing a spot matrix based on the sensing area; The point of the theoretical coordinate of the sensing area is locked, and the light spot matrix is ​​analyzed in the up, down, left, right, upper left, lower left, upper right, and lower right directions, and the light spot occupied area and the light spot contour line length in each direction are obtained respectively to obtain the first deviation coefficient of the corresponding direction; ； in, represents the first deviation coefficient in the i1th direction; Represents the theoretical spot area in the i1th direction; represents the area occupied by the light spot in the i1th direction; Indicates the length of the light spot contour line in the i1th direction; Indicates that the i1th direction is The theoretical length determined for the radius; is a constant, with a value of 3.14; Indicates the preset offset area in the i1th direction; ln indicates the sign of the logarithmic function; the value of i is 1, 2, 3, 4, 5, 6, 7, 8; All first deviation coefficients are divided according to the deviation level division table, and an initial analysis vector is constructed according to the number of occurrences of the first deviation coefficients involved in the same division level; Inputting the initial analysis vector into a deviation analysis model to obtain a reference error; According to the current setting combination of all filters of the flow cytometer and in combination with the light direction of the incident light source, a first analysis vector is constructed, and input into a forward analysis model to obtain first setting deviation information of each filter; According to the current setting combination of all filters of the flow cytometer and in combination with the light direction of the outgoing light source, a second analysis vector is constructed, and input into the reverse analysis model to obtain the second setting deviation information of each filter; Based on the first setting deviation information and the second setting deviation information, sequentially determining the self-light source error of each filter; According to the own light source error of each filter and combined with the reference error, an error setting function is constructed.

2. The method for calibrating a flow cytometer detector according to claim 1, characterized in that: According to the coordinate positioning of the sensing area of ​​the sensor panel, the position error d between the actual light source and the theoretical light source is calculated, including: Get the theoretical coordinates based on the sensing area ; Capture actual coordinates based on the sensing area ; Depends on the theoretical coordinates With actual coordinates , and obtain the position error d.

3. The method for calibrating a flow cytometer detector according to claim 2, characterized in that: Determine the deflection angle of the filter and the adjustment angle of the eccentric wheel, including: According to a first relationship expression between the optical filter and the position error d, a deflection angle of the optical filter is obtained; According to the second relationship expression between the optical filter and the eccentric wheel, the adjustment angle of the eccentric wheel is obtained.

4. The method for calibrating a flow cytometer detector according to claim 1, characterized in that: Determine the intrinsic light source error of each filter in turn, including: Extracting the first deviation of the corresponding filter from the first setting deviation information and the second setting deviation information respectively and the second deviation ; when When , the error of the own light source is determined to be 0; when When , the error of the own light source is determined to be r1; when ,according to Lock the minimum value, when the minimum value is When It is determined to be the self-light source error, where m0 represents the total number of filters; Indicates that all filters are based on the first deviation and the second deviation The variance of the absolute value of the difference; Indicates that all filters are based on the first deviation The variance of Indicates that all filters are based on the second deviation The variance of , and r1, r2 are variables; min represents the minimum value symbol; When the minimum value is When Determined to be self-light source error; When the minimum value is When It is determined to be the error of the own light source, where Indicates the first deviation of the jth filter and the second deviation The absolute value of the difference; when When the error is not zero, the error of the light source itself is determined.

5. The method for calibrating a flow cytometer detector according to claim 3, characterized in that: The control motor drives the eccentric wheel to correct the filter, including: Performing a first matching operation on the error setting function and the function-filter comparison table to obtain a first calibration solution, and performing a second matching operation on the error setting function and the function-eccentric wheel comparison table to obtain a second calibration solution; Based on the first calibration scheme and the second calibration scheme, respectively optimizing the deflection angle of the filter and the adjustment angle of the eccentric wheel; Based on the optimization result, the motor is controlled to drive the eccentric wheel to correct the filter.

6. The method for calibrating a flow cytometer detector according to claim 5, characterized in that: Based on the first calibration scheme and the second calibration scheme, the deflection angle of the filter and the adjustment angle of the eccentric wheel are optimized respectively, including: Extracting a first error angle for the optical filter from the first calibration scheme, and obtaining a second error angle based on the eccentric wheel according to a second relationship expression between the optical filter and the eccentric wheel; Extracting a third error angle for the eccentric wheel from the second calibration scheme, and obtaining a fourth error angle based on the filter according to a second relationship expression between the filter and the eccentric wheel; Taking the average value of the first error angle and the fourth error angle as the first optimized angle of the filter; Determining a second optimized angle for the eccentric wheel according to the first optimized angle, the second error angle, and the third error angle; Recording the sum of the adjustment angle and the second optimization angle, the third optimization angle converted from the first optimization angle based on the second relationship expression, and the adjustment angle, respectively, to control the first position point and the second position point reached by the eccentric wheel after the movement ends; Determine a first distance between the first position point and the theoretical coordinates, and at the same time, determine a second distance between the second position point and the theoretical coordinates; According to min (first distance, second distance), the corresponding final optimization angle is selected as an optimization reference.

7. The method for calibrating a flow cytometer detector according to claim 1, characterized in that: Also includes: Set deviation thresholds, including: Capture the moving path of the center point of the light spot on the sensor panel during the process of the control motor driving the eccentric wheel to correct the filter, divide the moving path equally to obtain several moving points, and determine the relationship between each moving point and the theoretical coordinates respectively Straight-line distance; Determine the absolute value of the distance difference between the two adjacent moving points according to the order of the moving points, and average all the absolute values ​​of the distance differences to obtain the unit difference. ; Obtaining the instrument attributes of the flow cytometer, and obtaining the initial threshold value yc0 from the attribute-threshold comparison table; Determine the detection accuracy Jd according to the experimental purpose of the cells to be detected; Based on the initial threshold yc0, the detection accuracy Jd and the unit difference , the absolute value of the distance difference between the last moving point and the second last moving point Ld, and the deviation threshold Py is obtained; ； in, represents the precision conversion function of the initial threshold yc0; n4 represents the number of moving points; Indicates the conversion factor between accuracy and threshold.

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