阅读说明：本技术 一种进行自动调节cv质控的方法、装置及相应的流式细胞仪 (Method and device for automatically adjusting CV quality control and corresponding flow cytometer ) 是由 饶云 封飘 胡钰静 周鹏 肖昌林 唐雪辉 于 2020-12-03 设计创作，主要内容包括：本发明公开了一种进行自动调节CV质控的方法、装置及相应的流式细胞仪,基于散点图中标准微球粒子的特征以及位置信息,自动形成矩形门,基于各通道直方图中标准微球粒子的特征以及位置信息,自动形成线性门,根据逐步逼近的算法自动分析计算合适的电压设置参数,从而对各通道的电压设置参数进行自动调整,并自动更新散点图与各通道直方图,使调整电压设置参数后的散点图中的标准微球粒子处于散点图中心与直方图中心的效果,使仪器达到最佳状态,从而使CV质控测试图形与门控调整、仪器控制的过程自动化,减少了用户工作量,提升了准确度,并且自动设置需要调节的仪器采集参数,使得对于用户的经验要求大大降低。(The invention discloses a method, a device and a corresponding flow cytometer for automatically adjusting CV quality control, wherein a rectangular gate is automatically formed based on the characteristics and the position information of standard microspherical particles in a scatter diagram, a linear gate is automatically formed based on the characteristics and the position information of the standard microspherical particles in histograms of all channels, and proper voltage setting parameters are automatically analyzed and calculated according to a successive approximation algorithm, so that the voltage setting parameters of all channels are automatically adjusted, the scatter diagram and the histograms of all channels are automatically updated, the standard microspherical particles in the scatter diagram after the voltage setting parameters are adjusted are positioned in the center of the scatter diagram and the center of the histograms, the instrument is in the best state, thereby the processes of CV quality control test graph and gate control adjustment and instrument control are automated, the workload of a user is reduced, the accuracy is improved, and the acquisition parameters of the instrument to be adjusted are automatically set, so that the experience requirements for the user are greatly reduced.)

1. A method for automatically adjusting CV quality control is characterized by comprising the following steps:

detecting whether the initial setting of the threshold of the acquisition parameter is normal or not;

detecting the initial voltage setting of each channel of the acquisition parameters, and judging whether the voltage setting meets the range;

sending a counting starting instruction to a lower computer, and starting a counting process by an instrument;

starting an automatic gate control adjustment algorithm subunit, and calculating the gate control position of each graph data;

calculating the median of standard microsphere particles in each channel histogram linear gate, judging whether the median in each channel histogram linear gate meets the median tolerance or not according to the median position standard of each channel specified by an instrument, and updating a flag bit whether the median of each channel passes or not;

starting an automatic acquisition parameter adjustment algorithm subunit, recalculating an acquisition parameter voltage value, and sending a new acquisition parameter value to a lower computer;

after the particle data are emptied, sending a re-acquisition instruction to a lower computer, redrawing a scatter diagram of a forward channel and a lateral channel and histogram data of each channel by software, and calculating a gating position of each graph data;

obtaining a gating position according to the histogram data of each channel by using an automatic gating adjustment algorithm, and calculating the number of particles in the linear gate of the histogram of each channel;

judging whether the algorithm reaches an iteration stop condition;

the software stops collecting data;

and automatically calculating the median of the histogram linear gate of each channel, the number of particles in the gate, the CV value and the median tolerance of each channel, judging whether the quality control result passes or not, and generating a quality control report.

2. The method for automatically adjusting CV quality control according to claim 1, wherein in detecting whether the threshold initialization setting of the acquisition parameter is normal or not, comprising the steps of,

judging whether the initial threshold setting of the acquisition parameters is in accordance with the normal condition, if so, continuously judging whether the initial voltage setting of each channel of the acquisition parameters is in accordance with the range; if not, resetting the initial value of the threshold value of the acquisition parameter, sending the initial value to the lower computer to reset the instrument parameter, and judging whether the initial voltage setting of each channel of the acquisition parameter meets the range.

3. The method for automatically adjusting CV quality control according to claim 1, wherein in the step of detecting the initial setting of the voltage of each channel for acquiring parameters and determining whether the range is met, the method comprises the steps of,

judging whether the initial voltage setting of each channel of the acquisition parameters is in accordance with the normal condition, and starting a counting process if the voltage setting ranges of the forward channel, the lateral channel and each fluorescence channel are in accordance with the normal setting range; if not, resetting the voltage initial value of the acquisition parameter of each channel, sending the voltage initial value to the lower computer to reset the instrument parameter, and starting the counting process.

4. The method for automatically adjusting CV quality control as set forth in claim 1, wherein the step of sending a start counting command to the lower computer, the start counting process of the instrument comprises,

after the counting process is started, software draws a scatter diagram of a forward channel and a lateral channel, the forward channel, the lateral channel and histograms of all fluorescence channels in real time; in particular, the automatic gating regulation algorithm subunit is activated when every fixed number of cells is added.

5. The method for automatically adjusting CV quality control according to claim 1, wherein in the step of starting the automatic gate control algorithm subunit and calculating the gate control position of each graph data, comprises the steps of,

reading data of an FSC-H forward channel and SSC-H side channel;

calculating the linear gate positions of histograms of an FSC-H forward channel and an SSC-H lateral channel;

calculating a rectangular gate position of a scatter diagram according to the linear gate positions of the histograms of the FSC-H forward channel and the SSC-H side channel;

screening data of each channel according to the rectangular door position of the scatter diagram;

linear gate positions are calculated for all height signal channel histograms.

6. The method for automatically adjusting CV quality control of claim 5, wherein in calculating histogram linear gate position, comprising the steps of,

screening each channel data through a rectangular door position of a scatter diagram, and converting the channel data into a histogram array;

solving the positions of the wave troughs on two sides of the maximum peak of the histogram, calculating the position of the maximum peak according to the histogram data, and searching and determining the positions of the wave trough on the left side of the maximum peak and the wave trough on the right side of the maximum peak by utilizing a downhill method;

and calculating the linear gate position of the histogram according to the trough positions on two sides of the maximum peak of the histogram.

7. The method for automatically adjusting CV quality control according to claim 1, wherein in the step of starting the automatic acquisition parameter adjustment algorithm subunit, recalculating the acquisition parameter voltage values and sending the new acquisition parameter values to the lower computer, comprises the steps of,

judging whether the initial value of the acquisition parameter conforms to a normal range or not, and judging whether the voltage setting of each channel of the acquisition parameter conforms to the range or not, wherein the voltage setting ranges comprise a forward channel, a lateral channel and each fluorescence channel;

correcting the forward FSC channel voltage;

judging whether the cell number reaches a threshold number for starting SSC voltage regulation;

correcting the SSC lateral channel voltage;

and correcting PMT voltage of each fluorescence channel.

8. The method for automatically adjusting CV quality control of claim 7, wherein in the "voltage modification", comprising the steps of,

judging whether the median in the linear gate of the channel histogram meets the median tolerance, if not, carrying out the next step, and if so, ending the process;

calculating the deviation amplitude and the deviation direction of the median value in the linear gate of the channel histogram;

according to the deviation amplitude and the deviation direction of the median value in the channel histogram linear gate, automatically increasing or decreasing the adjustment step length and the adjustment direction on the channel initial voltage parameter value by utilizing a successive approximation method to form a new current channel voltage parameter value;

judging whether the new voltage parameter value exceeds a normal range, if the new voltage parameter value is larger than the normal range, setting the new voltage value as the upper limit value of the normal voltage parameter range, and then performing the next step, if the new voltage parameter value is smaller than the normal range, setting the new voltage value as the lower limit value of the normal voltage parameter range, and then performing the next step, and if the new voltage parameter value is within the normal range, directly performing the next step;

and sending the new voltage value to a lower computer, resetting the acquisition parameters of the instrument, and finishing.

9. A device for automatically adjusting CV quality control is characterized by comprising a graph drawing unit, an instrument control unit, an automatic gate control adjusting unit, an automatic acquisition parameter adjusting unit and a report display unit; the graph drawing unit is used for drawing a scatter diagram and histograms of all channels of the forward channel and the lateral channel; the instrument control unit is used for automatically controlling the instrument; the automatic gate control adjusting unit is used for calculating the gate control position of each graphic data; the automatic acquisition parameter adjusting unit is used for recalculating the acquisition parameter voltage value and resetting the instrument parameters; the report display unit is used for generating a quality control report.

10. A flow cytometer comprising the means for performing automatically adjusted CV quality control of claim 9.

Technical Field

The invention relates to the field of medical equipment, in particular to a method and a device for automatically adjusting CV quality control and a corresponding flow cytometer.

Background

At present, a manual CV quality control test is adopted, which generally needs to adopt a plurality of tests to set voltage parameters of forward, lateral and each fluorescence channel, including establishing a scatter diagram and a histogram of the CV test, setting a graphic display range in a reasonable range, drawing a rectangular gate and a linear gate for calibration on the diagram, adjusting the position of a standard microspherical particle to an optimal position in the scatter diagram of the forward channel and the lateral channel, then setting the voltage parameters of each fluorescence channel, adjusting signal peaks on all fluorescence channel histograms to the optimal position, and besides, manually moving the position of the scatter rectangular gate during each acquisition, correctly enclosing the standard microspherical particle, manually moving the position of the linear gate on each channel histogram, enclosing the position of the signal peak, and calculating the number of cells of the signal peak of each channel histogram, And finally, judging whether the CV quality control test meets the standard or not according to the information such as the median value, the CV value and the like.

The manual CV test mode needs to adjust the voltage parameters, the gating positions and the like of each channel according to the experience of a tester, and retesting is needed after the parameters are adjusted every time, so that the steps are complicated, the accuracy is not enough, the instrument cannot reach the optimal state, and the resolution test result of the instrument is influenced.

Disclosure of Invention

The invention aims to provide a method and a device for automatically adjusting CV quality control and a corresponding flow cytometer, aiming at solving the technical problems that in the prior art, the mode of manually performing CV test needs to adjust the voltage parameter, the gating position and the like of each channel by depending on the experience of a tester, and retest is needed after each parameter adjustment, so that the steps are complicated, the accuracy is not enough, the instrument cannot reach the optimal state, and the resolution test result of the instrument is influenced.

In order to achieve the above object, the present invention provides a method for automatically adjusting CV quality control, comprising the steps of:

detecting whether the initial setting of the threshold of the acquisition parameter is normal or not;

detecting the initial voltage setting of each channel of the acquisition parameters, and judging whether the voltage setting meets the range;

sending a counting starting instruction to a lower computer, and starting a counting process by an instrument;

starting an automatic gate control adjustment algorithm subunit, and calculating the gate control position of each graph data;

calculating the median of standard microsphere particles in each channel histogram linear gate, judging whether the median in each channel histogram linear gate meets the median tolerance or not according to the median position standard of each channel specified by an instrument, and updating a flag bit whether the median of each channel passes or not;

starting an automatic acquisition parameter adjustment algorithm subunit, recalculating an acquisition parameter voltage value, and sending a new acquisition parameter value to a lower computer;

after the particle data are emptied, sending a re-acquisition instruction to a lower computer, redrawing a scatter diagram of a forward channel and a lateral channel and histogram data of each channel by software, and calculating a gating position of each graph data;

obtaining a gating position according to the histogram data of each channel by using an automatic gating adjustment algorithm, and calculating the number of particles in the linear gate of the histogram of each channel;

judging whether the algorithm reaches an iteration stop condition;

the software stops collecting data;

and automatically calculating the median of the histogram linear gate of each channel, the number of particles in the gate, the CV value and the median tolerance of each channel, judging whether the quality control result passes or not, and generating a quality control report.

Wherein, in the step of detecting whether the threshold initial setting of the acquisition parameters is in accordance with the normal condition,

judging whether the initial threshold setting of the acquisition parameters is in accordance with the normal condition, if so, continuously judging whether the initial voltage setting of each channel of the acquisition parameters is in accordance with the range; if not, resetting the initial value of the threshold value of the acquisition parameter, sending the initial value to the lower computer to reset the instrument parameter, and judging whether the initial voltage setting of each channel of the acquisition parameter meets the range.

Wherein, in the step of 'detecting the initial setting of the voltage of each channel of the acquisition parameters and judging whether the voltage meets the range',

judging whether the initial voltage setting of each channel of the acquisition parameters is in accordance with the normal condition, and starting a counting process if the voltage setting ranges of the forward channel, the lateral channel and each fluorescence channel are in accordance with the normal setting range; if not, resetting the voltage initial value of the acquisition parameter of each channel, sending the voltage initial value to the lower computer to reset the instrument parameter, and starting the counting process.

Wherein, in the process of sending a counting starting command to a lower computer and starting counting of an instrument,

after the counting process is started, software draws a scatter diagram of a forward channel and a lateral channel, the forward channel, the lateral channel and histograms of all fluorescence channels in real time; in particular, the automatic gating regulation algorithm subunit is activated when every fixed number of cells is added.

The method comprises the following steps of calculating the gating position of each graph data by a starting automatic gating regulation algorithm subunit:

reading data of an FSC-H forward channel and SSC-H side channel;

calculating the linear gate positions of histograms of an FSC-H forward channel and an SSC-H lateral channel;

calculating a rectangular gate position of a scatter diagram according to the linear gate positions of the histograms of the FSC-H forward channel and the SSC-H side channel;

screening data of each channel according to the rectangular door position of the scatter diagram;

linear gate positions are calculated for all height signal channel histograms.

Wherein in the "calculating histogram linear gate positions", the following steps are included,

screening each channel data through a rectangular door position of a scatter diagram, and converting the channel data into a histogram array;

solving the positions of the wave troughs on two sides of the maximum peak of the histogram, calculating the position of the maximum peak according to the histogram data, and searching and determining the positions of the wave trough on the left side of the maximum peak and the wave trough on the right side of the maximum peak by utilizing a downhill method;

and calculating the linear gate position of the histogram according to the trough positions on two sides of the maximum peak of the histogram.

Wherein, in the step of starting the automatic acquisition parameter adjustment algorithm subunit, recalculating the voltage value of the acquisition parameter and sending the new acquisition parameter value to the lower computer, the method comprises the following steps,

judging whether the initial value of the acquisition parameter conforms to a normal range or not, and judging whether the voltage setting of each channel of the acquisition parameter conforms to the range or not, wherein the voltage setting ranges comprise a forward channel, a lateral channel and each fluorescence channel;

correcting the forward FSC channel voltage;

judging whether the cell number reaches a threshold number for starting SSC voltage regulation;

correcting the SSC lateral channel voltage;

and correcting PMT voltage of each fluorescence channel.

Wherein, in the step of voltage correction, the following steps are included,

judging whether the median in the linear gate of the channel histogram meets the median tolerance, if not, carrying out the next step, and if so, ending the process;

calculating the deviation amplitude and the deviation direction of the median value in the linear gate of the channel histogram;

according to the deviation amplitude and the deviation direction of the median value in the channel histogram linear gate, automatically increasing or decreasing the adjustment step length and the adjustment direction on the channel initial voltage parameter value by utilizing a successive approximation method to form a new current channel voltage parameter value;

judging whether the new voltage parameter value exceeds a normal range, if the new voltage parameter value is larger than the normal range, setting the new voltage value as the upper limit value of the normal voltage parameter range, and then performing the next step, if the new voltage parameter value is smaller than the normal range, setting the new voltage value as the lower limit value of the normal voltage parameter range, and then performing the next step, and if the new voltage parameter value is within the normal range, directly performing the next step;

and sending the new voltage value to a lower computer, resetting the acquisition parameters of the instrument, and finishing.

A device for automatically adjusting CV quality control comprises a graph drawing unit, an instrument control unit, an automatic gate control adjusting unit, an automatic acquisition parameter adjusting unit and a report display unit; the graph drawing unit is used for drawing a scatter diagram and histograms of all channels of the forward channel and the lateral channel; the instrument control unit is used for automatically controlling the instrument; the automatic gate control adjusting unit is used for calculating the gate control position of each graphic data; the automatic acquisition parameter adjusting unit is used for recalculating the acquisition parameter voltage value and resetting the instrument parameters; the report display unit is used for generating a quality control report.

A flow cytometer comprising the means for performing automatically adjusted CV quality control of claim 9.

The invention relates to a method and a device for automatically adjusting CV quality control and a corresponding flow cytometer, wherein a rectangular gate is automatically formed based on the characteristics and the position information of standard microspherical particles in a scatter diagram, a linear gate is automatically formed based on the characteristics and the position information of the standard microspherical particles in histograms of all channels, and proper voltage setting parameters are automatically analyzed and calculated according to a successive approximation algorithm, so that the voltage setting parameters of all channels are automatically adjusted, the scatter diagram and the histograms of all channels are automatically updated, the standard microspherical particles in the scatter diagram after the voltage setting parameters are adjusted are positioned in the center of the scatter diagram and the center of the histograms, the instrument is in the best state, thereby the processes of CV quality control test graph and gate control adjustment and instrument control are automated, the workload of a user is reduced, the accuracy is improved, and instrument acquisition parameters needing to be adjusted are automatically set, so that the experience requirements for the user are greatly reduced.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.

FIG. 1 is a flow chart of a method of performing an automatically adjusting CV quality control of the present invention.

FIG. 2 is a flow chart of "one-touch quality control" according to the present invention.

FIG. 3 is a flow chart of an automatic gating adjustment algorithm subunit of the present invention.

FIG. 4 is a flow chart of an automatic acquisition parameter adjustment algorithm subunit of the present invention.

Fig. 5 is a flow chart of the automatic channel voltage successive approximation algorithm of the present invention.

FIG. 6 is an automatic CV quality control testing apparatus of the present invention.

FIG. 7 is a scatter plot of the present invention without acquisition parameter voltage adjustment.

FIG. 8 is a forward FSC-H channel histogram of the present invention without acquisition parameter voltage adjustment.

FIG. 9 is a FITC-H fluorescence channel histogram of the present invention without acquisition parameter voltage adjustment.

Fig. 10 is an acquisition parameter set window of the present invention without acquisition parameter voltage adjustment.

FIG. 11 is a comparison of scattergrams before and after performing an automatic CV quality control test according to the present invention.

FIG. 12 is a comparison of forward FSC-H histograms of the invention before and after performing an automatic CV quality control test.

FIG. 13 is a comparison of FITC-H fluorescence channel histograms before and after performing an automated CV quality control test in accordance with the present invention.

FIG. 14 is a window of acquisition parameter settings before and after performing an automatic CV quality control test in accordance with the present invention.

FIG. 15 is a diagram of quality control test report results in accordance with the present invention.

In the figure: 1-a graph drawing unit, 2-an instrument control unit, 3-an automatic gate control adjusting unit, 4-an automatic acquisition parameter adjusting unit, 5-a report display unit, 100-a device for automatically adjusting CV quality control and 200-a flow cytometer.

Detailed Description

Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the drawings are illustrative and intended to be illustrative of the invention and are not to be construed as limiting the invention.

Referring to fig. 1 to 15, the present invention provides a method for performing quality control of automatically adjusting CV, which includes the following steps:

s1001: detecting whether the initial setting of the threshold of the acquisition parameter is normal or not;

s1002: detecting the initial voltage setting of each channel of the acquisition parameters, and judging whether the voltage setting meets the range;

s1003: sending a counting starting instruction to a lower computer, and starting a counting process by an instrument;

s1004: starting an automatic gate control adjustment algorithm subunit, and calculating the gate control position of each graph data;

s1005: calculating the median of standard microsphere particles in each channel histogram linear gate, judging whether the median in each channel histogram linear gate meets the median tolerance or not according to the median position standard of each channel specified by an instrument, and updating a flag bit whether the median of each channel passes or not;

s1006: starting an automatic acquisition parameter adjustment algorithm subunit, recalculating an acquisition parameter voltage value, and sending a new acquisition parameter value to a lower computer;

s1007: after the particle data are emptied, sending a re-acquisition instruction to a lower computer, redrawing a scatter diagram of a forward channel and a lateral channel and histogram data of each channel by software, and calculating a gating position of each graph data;

s1008: obtaining a gating position according to the histogram data of each channel by using an automatic gating adjustment algorithm, and calculating the number of particles in the linear gate of the histogram of each channel;

s1009: judging whether the algorithm reaches an iteration stop condition;

s1010: the software stops collecting data;

s1011: and automatically calculating the median of the histogram linear gate of each channel, the number of particles in the gate, the CV value and the median tolerance of each channel, judging whether the quality control result passes or not, and generating a quality control report.

Further, in the step of detecting whether the threshold initial setting of the acquisition parameters is normal or not,

judging whether the initial threshold setting of the acquisition parameters is in accordance with the normal condition, if so, continuously judging whether the initial voltage setting of each channel of the acquisition parameters is in accordance with the range; if not, resetting the initial value of the threshold value of the acquisition parameter, sending the initial value to the lower computer to reset the instrument parameter, and judging whether the initial voltage setting of each channel of the acquisition parameter meets the range.

Further, in the step of 'detecting the initial setting of the voltage of each channel of the acquisition parameters and judging whether the voltage meets the range',

judging whether the initial voltage setting of each channel of the acquisition parameters is in accordance with the normal condition, and starting a counting process if the voltage setting ranges of the forward channel, the lateral channel and each fluorescence channel are in accordance with the normal setting range; if not, resetting the voltage initial value of the acquisition parameter of each channel, sending the voltage initial value to the lower computer to reset the instrument parameter, and starting the counting process.

Further, in "send start counting command to lower computer, instrument start counting procedure",

after the counting process is started, software draws a scatter diagram of a forward channel and a lateral channel, the forward channel, the lateral channel and histograms of all fluorescence channels in real time; in particular, the automatic gating regulation algorithm subunit is activated when every fixed number of cells is added.

Further, in the sub-unit for starting the automatic gate control adjustment algorithm and calculating the gate control position of each graphic data, the method comprises the following steps:

reading data of an FSC-H forward channel and SSC-H side channel;

calculating the linear gate positions of histograms of an FSC-H forward channel and an SSC-H lateral channel;

calculating a rectangular gate position of a scatter diagram according to the linear gate positions of the histograms of the FSC-H forward channel and the SSC-H side channel;

screening data of each channel according to the rectangular door position of the scatter diagram;

linear gate positions are calculated for all height signal channel histograms.

Further, in "calculating histogram linear gate position", the steps are included,

screening each channel data through a rectangular door position of a scatter diagram, and converting the channel data into a histogram array;

solving the positions of the wave troughs on two sides of the maximum peak of the histogram, calculating the position of the maximum peak according to the histogram data, and searching and determining the positions of the wave trough on the left side of the maximum peak and the wave trough on the right side of the maximum peak by utilizing a downhill method;

and calculating the linear gate position of the histogram according to the trough positions on two sides of the maximum peak of the histogram.

Furthermore, in the step of starting the automatic acquisition parameter adjustment algorithm subunit, recalculating the voltage value of the acquisition parameter and sending the new value of the acquisition parameter to the lower computer, the method comprises the following steps,

judging whether the initial value of the acquisition parameter conforms to a normal range or not, and judging whether the voltage setting of each channel of the acquisition parameter conforms to the range or not, wherein the voltage setting ranges comprise a forward channel, a lateral channel and each fluorescence channel;

correcting the forward FSC channel voltage;

judging whether the cell number reaches a threshold number for starting SSC voltage regulation;

correcting the SSC lateral channel voltage;

and correcting PMT voltage of each fluorescence channel.

Further, in the "voltage correction", the following steps are included,

judging whether the median in the linear gate of the channel histogram meets the median tolerance, if not, carrying out the next step, and if so, ending the process;

calculating the deviation amplitude and the deviation direction of the median value in the linear gate of the channel histogram;

according to the deviation amplitude and the deviation direction of the median value in the channel histogram linear gate, automatically increasing or decreasing the adjustment step length and the adjustment direction on the channel initial voltage parameter value by utilizing a successive approximation method to form a new current channel voltage parameter value;

judging whether the new voltage parameter value exceeds a normal range, if the new voltage parameter value is larger than the normal range, setting the new voltage value as the upper limit value of the normal voltage parameter range, and then performing the next step, if the new voltage parameter value is smaller than the normal range, setting the new voltage value as the lower limit value of the normal voltage parameter range, and then performing the next step, and if the new voltage parameter value is within the normal range, directly performing the next step;

and sending the new voltage value to a lower computer, resetting the acquisition parameters of the instrument, and finishing.

Further, a device 100 for automatically adjusting CV quality control comprises a graph drawing unit 1, an instrument control unit 2, an automatic gate control adjusting unit 3, an automatic acquisition parameter adjusting unit 4 and a report display unit 5; the graph drawing unit 1 is used for drawing a scatter diagram and a histogram of a forward channel and a lateral channel; the instrument control unit 2 is used for automatically controlling an instrument; the automatic gate control adjusting unit 3 is used for calculating the gate control position of each graphic data; the automatic acquisition parameter adjusting unit 4 is used for recalculating the acquisition parameter voltage value and resetting the instrument parameters; the report display unit 5 is used for generating a quality control report.

Further, a flow cytometer 200 comprising the apparatus 100 for performing automatically adjusting CV quality control of claim 9.

In the present embodiment, for convenience of the following description, some terms referred to hereinafter need to be briefly described as follows:

flow data refers to data obtained by irradiating fluorescent dye on a measured object by laser and collecting signal intensity of scattered light and fluorescence excitation light at each angle by using flow cytometry;

a scattergram which is a two-dimensional map generated by a flow cytometer and on which two-dimensional characteristic information of a plurality of particles is distributed, wherein an X coordinate axis and a Y coordinate axis of the scattergram each represent a characteristic of each particle, for example, in a scattergram, the X coordinate axis represents a size characteristic and the Y coordinate axis represents an internal complexity characteristic of a particle;

a histogram which is a one-dimensional graph generated by a flow cytometry and on which one-dimensional characteristic information of a plurality of particles is distributed, wherein an X coordinate axis of the histogram represents a characteristic cell group of each particle;

a histogram peak, which is a particle peak formed of a plurality of particles having the same characteristic, concentrated in a certain region of the histogram;

in the embodiment of the invention, the following ideas are adopted:

when the standard microspheroidal particle flow data generated by the flow cytometer 200 needs to be analyzed, a user can select to manually adjust the voltage value in the interactive interface to perform the CV quality control test, or automatically perform the CV quality control test by the "one-key quality control" method provided by the embodiment of the present invention. In the scattergram shown in fig. 7 without voltage adjustment, the standard microspheroidal particle is not located at the center of the scattergram, in the forward FSC-H channel histogram without voltage adjustment shown in fig. 8, the standard microspheroidal particle peak is not located at the center of the histogram, in the FITC-H fluorescence channel histogram without voltage adjustment for the acquisition parameter shown in fig. 9, the standard microspheroidal particle peak is not located at the center of the histogram, and in fig. 10, the acquisition parameter setting window without voltage adjustment for the acquisition parameter is shown.

A user can select to set voltage parameter values of each channel in a parameter acquisition setting window, the current voltage value of the FSC channel in the graph 10 is 559, the user can click a 'running' button, manually adjust the voltage value as required while an instrument collects data, manually click a're-acquisition' button, re-collect data, and then manually adjust the position of a rectangular gate on a scatter diagram and the position of a linear gate on a histogram of each channel; or the user can select the quality control automatic test and click the 'one-key quality control' button in the graph, the method provided by the embodiment of the invention can automatically form a rectangular gate based on the characteristics and the position information of the standard microsphere particles in the histogram of each channel, automatically form a linear gate based on the characteristics and the position information of the standard microsphere particles in the histogram of each channel, automatically analyze and calculate the proper voltage setting parameters according to the successive approximation algorithm, thereby automatically adjusting the voltage setting parameters of each channel, automatically resetting the instrument parameters and re-collecting the particles, automatically updating the histogram of each channel and the histogram of each channel, enabling the standard microsphere particles in the histogram of each channel after adjusting the voltage setting parameters to be positioned in the center of the histogram and the center of the histogram, and automatically calculating the median, the number of particles in the gate, the CV value and the median tolerance of each channel in the linear gate of each channel, And the voltage tolerance of the voltage set value of each channel and the standard voltage set value automatically judges whether the quality control result passes or not, and generates a quality control report. After the 'one-key quality control' process is carried out, if a user considers that the automatically calculated channel voltage parameter value or the rectangular gate position on the scatter diagram is not appropriate to the linear gate position on the histogram, the user can click a 'run' button, manually adjust the voltage value according to the requirement while the instrument collects data, manually click a 'reacquisition' button, re-collect the data, and then manually adjust the position of the rectangular gate on the scatter diagram and the position of the linear gate on the histogram of each channel. The following mainly describes the principle and process of "one-key quality control" in the embodiment of the present invention:

judging whether the initial threshold setting of the acquisition parameters is in accordance with the normal condition, wherein the initial threshold setting comprises whether a channel option of a first threshold is correct, whether a set value of the first threshold is in accordance with a normal range, whether a channel option of a second threshold is correct, whether a set value of the second threshold is in accordance with the normal range, and whether the logical relationship between the first threshold and the second threshold is correct, if so, continuing to carry out the step 2; if not, resetting the initial value of the acquisition parameter threshold, sending the initial value to the lower computer to reset the instrument parameters, and continuing the step 2; 2. judging whether the initial voltage setting of each channel of the acquisition parameters meets the range, including the voltage setting ranges of the forward channel, the lateral channel and each fluorescence channel, and continuing the step 3 if the voltage setting ranges of the forward channel, the lateral channel and each fluorescence channel all meet the normal setting range; if not, resetting the voltage initial value of the acquisition parameter of each channel, sending the voltage initial value to the lower computer to reset the instrument parameter, and continuing to the step 3; 3. starting a counting process, sending a counting starting instruction to a lower computer, starting the counting process by an instrument, transmitting the acquired data to a software system by the lower computer after the sheath flow is stable, drawing a scatter diagram of a forward channel and a lateral channel, the forward channel, the lateral channel and histograms of all fluorescence channels by the software in real time, and continuing to step 4; 4. starting an automatic gate control adjustment algorithm subunit, when the number of particles is increased by a fixed number, starting the automatic gate control adjustment algorithm subunit to calculate the gate control position of each graphic data, firstly identifying the position of a standard microsphere particle in a scatter diagram, calculating the coordinate position of a rectangular gate, enclosing the standard microsphere particle in the rectangular gate, and drawing data of a histogram as the enclosed standard microsphere particle in the rectangular gate of the scatter diagram, then calculating the linear gate position of each histogram according to the position of the standard microsphere particle peak in each histogram, containing the standard microsphere particle peak in the linear gate, and continuing to the step 5; 5. calculating the median of standard microspherical particles in each channel histogram linear gate, judging whether the median in each channel histogram linear gate meets the median tolerance according to the median position standard of each channel specified by an instrument, updating a flag bit whether the median of each channel passes, if the median in each histogram linear gate meets the median tolerance of each channel, the flag bits of the median tolerance of all the channels pass, continuing to step 8, if only one channel does not meet the median tolerance of the histogram, setting the flag bit of the channel meeting the median tolerance as pass, and setting the rest channels not meeting the requirement of the median tolerance as fail, and continuing to step 6; 6. starting an automatic acquisition parameter adjusting algorithm subunit, recalculating the acquisition parameter voltage values according to the median tolerance between the last iteration acquisition parameter voltage values and the linear gate of each channel histogram, sending the new acquisition parameter values to a lower computer, resetting the instrument parameters, and continuing to step 7; 7. after the particle data are emptied, sending a re-acquisition instruction to a lower computer, redrawing a scatter diagram of the forward channel and the lateral channel and histogram data of each channel by software, and continuing to the step 4; 8. judging whether the number of particles in each histogram linear gate meets the number requirement, calculating the number of particles in the histogram linear gate of each channel according to the histogram data of each channel by utilizing the gating position obtained by an automatic gating adjustment algorithm, and continuing the step 10 if the number of particles in the histogram linear gate of each channel meets the requirement, or continuing the step 9 if the number of particles in the histogram linear gate of one channel does not meet the requirement; 9. judging whether the algorithm reaches an iteration stop condition, wherein the iteration stop condition of the whole 'one-key quality control' algorithm comprises an upper iteration number limit of the algorithm, a total particle collection number and particle collection time, the first iteration stop condition is that after the iteration number of the algorithm reaches the upper limit, the median tolerance flag positions of all channels are passed, the automatic collection parameter adjustment algorithm subunit is not started to reset collection parameters, meanwhile, after the total particle collection number reaches the requirement, the iteration is stopped, the second iteration stop condition is that after the particle collection time reaches the upper limit, the median tolerance flag positions of all channels are passed, the automatic collection parameter adjustment algorithm subunit is not started to reset collection parameters, and after the total particle collection number reaches the requirement, the iteration is stopped, if the algorithm meets one of the two stop conditions, the step 10 is continued, otherwise, continuing the step 4; 10. sending a counting stopping instruction to a lower computer, finishing an algorithm iteration flow, stopping collecting particles by an instrument, stopping collecting data by software, and continuing to step 11; 11. and automatically judging the quality control result by software to generate a quality control report, automatically calculating the median, the in-gate particle number and the CV value of the histogram linear gate of each channel and the median tolerance of each channel according to the data of the forward channel and each fluorescence channel, automatically judging whether the quality control result passes or not according to the median, the in-gate particle number and the CV value of the histogram linear gate of each channel and the median tolerance of each channel and the voltage tolerance of the voltage set value and the standard voltage set value of each channel, generating the quality control report, and ending the 'one-key quality control' process.

Further, the automatic gate control adjustment algorithm subunit in step 4 calculates the gate control position of each graphic data, and includes the following steps:

reading data of an FSC-H forward channel and SSC-H side channel;

calculating the linear gate positions of histograms of an FSC-H forward channel and an SSC-H lateral channel;

calculating a rectangular gate position of a scatter diagram according to the linear gate positions of the histograms of the FSC-H forward channel and the SSC-H side channel;

screening data of each channel according to the rectangular door position of the scatter diagram;

calculating the linear door positions of all height signal channel histograms;

still further, in the step (2) and the step (5), a histogram linear gate position algorithm is used, which includes the following steps:

screening data of each channel through a rectangular door position of a scatter diagram, and converting the data of the channel into a histogram array;

solving the positions of the wave troughs on two sides of the maximum peak of the histogram, calculating the position of the maximum peak according to the histogram data, and searching and determining the positions of the wave trough on the left side of the maximum peak and the wave trough on the right side of the maximum peak by utilizing a downhill method;

calculating the linear gate position of the histogram according to the trough positions on two sides of the maximum peak of the histogram;

further, the automatic acquisition parameter adjusting algorithm subunit in the "one-key quality control" step 6 calculates the acquisition parameter voltage value of each channel according to the acquisition parameter voltage value of each channel of the last iteration and the median position and the median tolerance in the linear gate of the histogram of each channel, and includes the following steps:

judging whether the initial values of the acquisition parameters conform to a normal range or not, judging whether the initial voltage settings of all channels of the acquisition parameters conform to ranges including voltage setting ranges of a forward channel, a lateral channel and all fluorescence channels, and continuing the step (2) if the voltage setting ranges of the forward channel, the lateral channel and all the fluorescence channels conform to the normal setting ranges; if not, resetting the voltage initial value of the acquisition parameter of each channel, sending the voltage initial value to the lower computer to reset the instrument parameter, and continuing the step (2);

correcting the forward FSC channel voltage;

judging whether the cell number reaches the threshold number for starting SSC voltage regulation, if so, continuing the step (4), and if not, continuing the step (5);

correcting the SSC lateral channel voltage;

correcting PMT voltage of each fluorescence channel;

furthermore, in the step (2), the step (4) and the step (5), the automatic channel voltage correction uses an automatic channel voltage successive approximation algorithm, which includes the following steps:

judging whether the median in the linear gate of the channel histogram meets the median tolerance, if not, continuing the step II, and if so, ending the process;

calculating the deviation amplitude and the deviation direction of the median value in the linear gate of the channel histogram;

thirdly, automatically increasing and decreasing the adjusting step length and the adjusting direction on the initial voltage parameter value of the channel by utilizing a successive approximation method according to the deviation amplitude and the deviation direction of the median value in the linear gate of the histogram of the channel to form a new current voltage parameter value of the channel;

judging whether the new voltage parameter value exceeds the normal range, if the new voltage parameter value is greater than the normal range, setting the new voltage value as the upper limit value of the normal voltage parameter range, and continuing the step, if the new voltage parameter value is less than the normal range, setting the new voltage value as the lower limit value of the normal voltage parameter range, and continuing the step, if the new voltage parameter value is within the normal range, directly continuing the step;

and fifthly, sending the new voltage value to a lower computer, resetting the acquisition parameters of the instrument, and ending.

It will be appreciated that in other embodiments, the successive approximation algorithm may employ, for example, a binary search algorithm.

After the "one-key quality control" process of the automatic CV quality control test is finished, in the scattergram after the automatic CV quality control test shown in fig. 11, the standard microsphere particle is located at the center of the scattergram, and in the forward FSC-H channel histogram after the automatic CV quality control test shown in fig. 12, the standard microsphere particle peak is located at the center of the histogram, for example, the FITC-H fluorescence channel histogram after the automatic CV quality control test shown in fig. 13, and the standard microsphere particle peak is located at the center of the histogram.

Fig. 14 shows the values of the channel voltage parameters in the acquisition parameter setting window after the automatic CV quality control test as the automatically adjusted voltage values, and fig. 15 shows the quality control result report after the automatic CV quality control test.

It will be appreciated that after automatically calculating the gating position and the values of the respective channel voltage parameters, the user may also manually adjust the gating position and the values of the channel voltage parameters if the user deems the automatically calculated gating position or the automatically calculated channel voltage parameters are not appropriate.

Correspondingly, in another aspect of the embodiment of the present invention, a device for performing an automatic CV quality control test is further provided, and includes the aforementioned graph drawing unit 1, the instrument control unit 2, the automatic gate control adjusting unit 3, the automatic acquisition parameter adjusting unit 4, and the report display unit 5.

Accordingly, in another aspect of the embodiments of the present invention, there is provided a flow cytometer 200 including the aforementioned apparatus for performing automatic CV quality control testing.

The method, the device and the corresponding flow cytometer 200 for performing automatic CV quality control testing provided by the embodiments of the present invention automatically form a rectangular gate based on the characteristics and the position information of the standard microspherical particles in the histogram of each channel, automatically form a linear gate based on the characteristics and the position information of the standard microspherical particles in the histogram of each channel, automatically analyze and calculate appropriate voltage setting parameters according to a successive approximation algorithm, thereby automatically adjusting the voltage setting parameters of each channel, automatically update the histogram of each channel and the histogram of each channel, enable the standard microspherical particles in the histogram of each channel after adjusting the voltage setting parameters to be in the center of the histogram and the center of the histogram, enable the instrument to reach an optimal state, automatically calculate the median, the number of particles in the linear gate, the CV value, the median tolerance of each channel, and the voltage tolerance of the voltage setting value and the standard voltage setting value of each channel, and automatically judging whether the quality control result passes or not and generating a quality control report.

The embodiment of the invention automates the CV quality control test pattern, gate control adjustment and instrument control processes, reduces the workload of users, improves the accuracy, and automatically sets the acquisition parameters of the instruments to be adjusted, thereby greatly reducing the experience requirements of the users.

While the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.