CN109580550A - A kind of classification processing method and its device of leucocyte - Google Patents
A kind of classification processing method and its device of leucocyte Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/47—Scattering, i.e. diffuse reflection
- G01N21/49—Scattering, i.e. diffuse reflection within a body or fluid
Abstract
The present invention discloses the classification processing method and its device of a kind of leucocyte.Method is comprising steps of obtain the first, second, third angle scattered light signal of each cell particle in leucocyte blood sample by light scattering testing technology；It is respectively horizontal axis, the main scatter plot of longitudinal axis drafting with second, first angle scattered light signal, third, second angle scattered light signal are respectively horizontal axis, longitudinal axis drafting auxiliary scatter plot；Preliminary classification is carried out to main scatter plot by clustering, judges whether preliminary classification information is effective；Effectively, then it performs the next step；In vain, then preliminary classification is carried out to main scatter plot by projective transformation, obtains effective preliminary classification information, then perform the next step；Using the cluster analysis result of auxiliary scatter plot, effective preliminary classification information is modified, the true classification information of leucocyte is obtained.The classification processing method of leucocyte of the invention improves the accuracy of leukocyte differential count, and accuracy is high, universality is good.
Description
Technical field
The present invention relates to blood cell differential technical field more particularly to the classification processing methods and its dress of a kind of leucocyte
It sets.
Background technique
Cellanalyzer is a kind of a kind of instrument of cell in detectable blood, can be to the leucocyte, red thin in blood
Born of the same parents, blood platelet etc. count and classify.Cellanalyzer realizes that a kind of most common technology of leukocyte differential count is laser
Scattering method.Blood sample is after reagent acts on, cell particle by illumination jet stream through detection zone, collects various types of cells grain
The scattered light signal of the different angle of son forms scatter plot, by carrying out clustering to scatter plot, to realize to leucocyte
Five classification.
Existing leucocyte classification method is usually to collect each cell particle by two generated when sheath flow pool different angles
The scattered light signal of degree, small angle scattering optical signal and middle angle scattered light signal, small angle scattering optical signal indicate cell
Size information, middle angle scattered light signal indicate the complexity (i.e. the grown form information of cell) of cell interior, Yi Zhongjiao
Scattered light signal is spent as horizontal axis, and small angle scattering optical signal draws two dimension as a point as the longitudinal axis, each cell particle
In-low-angle scatter plot, referred to as main scatter plot.By carrying out clustering to above-mentioned main scatter plot, point of leucocyte is obtained
Class, for common leucocyte blood sample, classification results as shown in Figure 1, specifically: 1 be monocyte, and 2 are
Property granulocyte, 3 be eosinophil, and 4 be lymphocyte, 5,6 be shadow cell；Wherein, eosinophil is made by reagent
With rear, since its internal complexity is higher, and volume be not it is very big, so its lower right corner for being distributed in main scatter plot；Meanwhile by
There are cell fragment and blood platelet etc. or the reasons such as erythroblast or platelet aggregation in blood sample, meeting exists
The lower left corner of main scatter plot forms shadow cell.And for some special leucocyte blood samples, shadow cell region
It can extend to upper right, mix with eosinophil region, to influence existing leucocyte classification method to acidophil granules
The Accurate classification of cell causes eosinophil classification results inaccurate；For other special leucocyte blood samples
For, it is existing without apparent boundary between neutrophil leucocyte and monocyte on main scatter plot after reagent acts on
Leucocyte classification method can not accurately distinguish neutrophil leucocyte and monocyte.
Therefore, the existing technology needs to be improved and developed.
Summary of the invention
In view of above-mentioned deficiencies of the prior art, the purpose of the present invention is to provide a kind of method for sorting leucocyte and its dresses
It sets, it is intended to it is inaccurate to some special leucocyte blood sample classification, universality difference to solve existing leucocyte classification method
Problem.
Technical scheme is as follows:
A kind of classification processing method of leucocyte, wherein comprising steps of
A, the red blood cell in whole blood sample is completely dissolved using reagent, obtains leucocyte blood sample；
B, by light scattering testing technology obtain leucocyte blood sample in each cell particle first angle scattered light signal,
Second angle scattered light signal and third angle scattered light signal；
It C, is that the longitudinal axis draws main scatter plot using second angle light scattering signal as horizontal axis, first angle scattered light signal, with third
Angle scattered light signal is horizontal axis, second angle scattered light signal is that the longitudinal axis draws auxiliary scatter plot；
D, preliminary classification is carried out to main scatter plot by clustering, judges whether preliminary classification information is effective；
Preliminary classification information is effective, thens follow the steps E；
Preliminary classification information is invalid, then carries out preliminary classification to main scatter plot by projective transformation, obtain effective preliminary classification
Information, then execute step E；
E, using the cluster analysis result to auxiliary scatter plot, the effective preliminary classification information is modified, is obtained white
The true classification information of cell.
The classification processing method of the leucocyte, wherein in step A, the reagent is hemolytic agent.
The classification processing method of the leucocyte, wherein in step B, the first angle scattered light signal represents thin
The information of born of the same parents' size, the second angle scattered light signal represent the essential information of cellular morphology, and the third angle scatters light
Signal represents the auxiliary information of cellular morphology.
The classification processing method of the leucocyte, wherein in step B, the first angle scattered light signal is corresponding
Angle is 1 ~ 6 °；The corresponding angle of the second angle scattered light signal is 6 ~ 18 °；The third angle scattered light signal is corresponding
Angle be 26 ~ 42 °.
The classification processing method of the leucocyte, wherein described that main scatter plot is tentatively divided by projective transformation
Class specifically includes step:
D1, projective transformation is carried out to its X direction and y direction to the point on main scatter plot respectively, it is straight to obtain two projections
Fang Tu；
D2, the disposal of gentle filter then is carried out to projection histogram, obtains smooth histogram；
Main scatter plot is carried out preliminary classification by D3, the threshold value using the wave trough position of the smooth histogram as segmentation.
The leukocyte differential count processing method, wherein the method that the disposal of gentle filter is carried out to projection histogram
For gaussian filtering method.
The classification processing method of the leucocyte, wherein the step E is specifically included:
E1, clustering is carried out to the auxiliary scatter plot, obtains the correcting region of certain class cell in leucocyte；
E2, the point for the correcting region that preliminary classification in main scatter plot does not fall within certain class cell for certain described class cell is picked
It removes, remaining point is the true group minute of certain class cell.
The method for sorting leucocyte, wherein certain described class cell is eosinophil or monocyte.
A kind of device of the classification processing method of leucocyte as described above, wherein include:
Fluid driving unit, for driving leucocyte blood sample；
Flow pool unit, for drive the cell particle in leucocyte blood sample individually sequentially through；
Light illuminating unit, for irradiating the cell particle for sequentially flowing through flowing pool unit；
Optical measurement unit, for measuring and obtaining the scattered light signal of cell particle；
The optical measurement unit includes:
First diaphragm, for measuring the first angle scattered light signal of cell particle；
Second diaphragm, for measuring the second angle scattered light signal of cell particle；
Third diaphragm, for measuring the third angle scattered light signal of cell particle；
First optical signal receiver, for obtaining the first angle scattered light signal and second angle scattering light letter of cell particle
Number；
Second optical signal receiver, for obtaining the second angle scattered light signal of cell particle；
Third optical signal receiver, for obtaining the third angle scattered light signal of cell particle.
The utility model has the advantages that the present invention collects third angle scattered light signal by increasing, and second angle scattered light signal is
The longitudinal axis, third angle scattered light signal are that horizontal axis draws auxiliary scatter plot, followed by the cluster analysis result of auxiliary scatter plot
The preliminary classification result of main scatter plot is modified, the accuracy of leukocyte differential count is improved；And leucocyte of the invention
The universality of classification processing method is good.
Detailed description of the invention
Fig. 1 is the classification results of leucocyte blood sample common in the prior art.
Fig. 2 is the structural schematic diagram of the device of the classification processing method of leucocyte in the specific embodiment of the invention.
Fig. 3 is the preliminary classification result of eosinophil in the leucocyte blood sample of the embodiment of the present invention 1.
Fig. 4 is the correcting region of eosinophil in the leucocyte blood sample of the embodiment of the present invention 1.
Fig. 5 is the true classification results of eosinophil in the leucocyte blood sample of the embodiment of the present invention 1.
Fig. 6 is the main scatter plot of the leucocyte blood sample of the embodiment of the present invention 2.
Fig. 7 is the projection histogram of the leucocyte blood sample of the embodiment of the present invention 2.
Fig. 8 is the preliminary classification result of monocyte in the leucocyte blood sample of the embodiment of the present invention 2.
Fig. 9 is the correcting region of monocyte in the leucocyte blood sample of the embodiment of the present invention 2.
Figure 10 is the true classification results of the monocyte in the leucocyte blood sample of the embodiment of the present invention 2.
Specific embodiment
The present invention provides a kind of classification processing method of leucocyte, to make the purpose of the present invention, technical solution and effect more
Add clear, clear, the present invention is described in more detail below.It should be appreciated that specific embodiment described herein is only used
To explain the present invention, it is not intended to limit the present invention.
The embodiment of the present invention provides a kind of classification processing method of leucocyte, wherein comprising steps of
A, the red blood cell in whole blood sample is completely dissolved using reagent, obtains leucocyte blood sample；
B, by light scattering testing technology obtain leucocyte blood sample in each cell particle first angle scattered light signal,
Second angle scattered light signal and third angle scattered light signal；
It C, is that the longitudinal axis draws main scatter plot using second angle light scattering signal as horizontal axis, first angle scattered light signal, with third
Angle scattered light signal is horizontal axis, second angle scattered light signal is that the longitudinal axis draws auxiliary scatter plot；
D, preliminary classification is carried out to main scatter plot by clustering, judges whether preliminary classification information is effective；
Preliminary classification information is effective, thens follow the steps E；
Preliminary classification information is invalid, then carries out preliminary classification to main scatter plot by projective transformation, obtain effective preliminary classification
Information, then execute step E；
E, using the cluster analysis result of auxiliary scatter plot, the effective preliminary classification information is modified, is obtained white thin
The true classification information of born of the same parents.
It should be noted that the angle formed according to the scattering light and the horizontal direction that obtain cell particle is of different sizes,
First angle scattered light signal corresponds to small angle scattering optical signal in the present embodiment, during second angle scattered light signal corresponds to
Angle scattered light signal, third angle scattered light signal correspond to large angle scattering optical signal.The present embodiment is represented by obtaining
The third angle scattered light signal of the auxiliary information of cellular morphology, with third angle scattered light signal and the base for representing cellular morphology
The second angle scattered light signal of this information draws the second-third angle scatter plot (auxiliary scatter plot)；And utilize auxiliary scatterplot
Figure is modified the classification results of main scatter plot, to improve the accuracy of leukocyte differential count.
In a preferred embodiment, in step A, the reagent can be but be not limited to hemolytic agent.
In a preferred embodiment, in step B, the first angle scattered light signal represents the letter of cell size
Breath, the second angle scattered light signal represent the essential information of cellular morphology, and the third angle scattered light signal represents thin
The auxiliary information of born of the same parents' form.
In a preferred embodiment, in step B, the corresponding angle of the first angle scattered light signal is 1 ~
6°；The corresponding angle of the second angle scattered light signal is 6 ~ 18 °；The corresponding angle of the third angle scattered light signal is
26~42°。
In a preferred embodiment, described that main scatter plot progress preliminary classification is specifically included by projective transformation
Step:
D1, projective transformation is carried out to its X direction and y direction to the point on main scatter plot respectively, it is straight to obtain two projections
Fang Tu；
D2, the disposal of gentle filter then is carried out to projection histogram, obtains smooth histogram；
Main scatter plot is carried out preliminary classification by D3, the threshold value using the wave trough position of the smooth histogram as segmentation.
In a preferred embodiment, the method that described pair of main scatter plot carries out the disposal of gentle filter is gaussian filtering
Method.
In a preferred embodiment, the step E is specifically included:
E1, clustering is carried out to the auxiliary scatter plot, obtains the correcting region of certain class cell in leucocyte；
E2, the point for the correcting region that preliminary classification in main scatter plot does not fall within certain class cell for certain described class cell is picked
It removes, remaining point is the true group minute of certain class cell.
In a preferred embodiment, certain described class cell is eosinophil or monocyte.
As shown in Fig. 2, the embodiment of the present invention provides a kind of device of the classification processing method of leucocyte as described above,
In, comprising:
Fluid driving unit H, for driving leucocyte blood sample；
Flow pool unit I, for drive the cell particle in leucocyte blood sample individually sequentially through；
Light illuminating unit J, for irradiating the cell particle for sequentially flowing through flowing pool unit；
Optical measurement unit K, for measuring and obtaining the scattered light signal of cell particle；
The optical measurement unit K includes:
First diaphragm 50a, for measuring the first angle scattered light signal of cell particle；
Second diaphragm 50b, for measuring the second angle scattered light signal of cell particle；
Third diaphragm 50c, for measuring the third angle scattered light signal of cell particle；
First optical signal receiver 60a, for obtaining the first angle scattered light signal of cell particle；
Second optical signal receiver 60b, for obtaining the second angle scattered light signal of cell particle
Third optical signal receiver 60c, for obtaining the third angle scattered light signal of cell particle.
Specifically, the fluid driving unit H of the device of the classification processing method of the leucocyte of the present embodiment, flowing pool unit
I, light illuminating unit J and the corresponding unit setting in common blood cell differential device are almost the same.Wherein, the illumination list of the present embodiment
It is laser light source that first J successively includes: 10 from left to right, using argon ion laser, to provide monochromatic light；20 be convex lens；
30a, 30b are respectively the first collimating mirror, the second collimating mirror.The flowing pool unit I of the present embodiment is sheath flow pool, to guarantee that cell is mixed
Suspension forms the cell stream individually arranged in detection liquid stream.Optical signal receiver in the optical measurement unit K of the present embodiment
For photodiode, to obtain the scattered light signal generated after laser irradiation cell particle.The driving of fluid driving unit is white thin
Born of the same parents' blood sample, the cell particle in sample is individually sequentially through flowing pool unit (sheath flow pool), the irradiation through light illuminating unit,
Optical measurement unit obtains scattered light signal.It should be noted that third angle scattered light signal and second angle scattering light letter
Number indicate the inside complexity of cell, it is only variant in degree；In the present embodiment, in order to distinguish, by second angle
Scattered light signal indicates that the inside complexity of cell is known as the essential information of cellular morphology, by third angle scattered light signal table
Show that the inside complexity of cell is known as the auxiliary information of cellular morphology.
In the present embodiment, by increasing third angle diaphragm, detection in the optical measurement unit of blood cell differential device
The third angle scattered light signal of the auxiliary information of cellular morphology is represented, with third angle scattered light signal and represents cellular morphology
The second angle scattered light signal of essential information draw the second-third angle scatter plot (auxiliary scatter plot)；And utilize auxiliary
Scatter plot is modified the classification results of main scatter plot, to improve the accuracy of leukocyte differential count.
Embodiment 1
At room temperature, the red blood cell in one whole blood sample is completely dissolved using hemolytic agent, obtains a leucocyte blood sample；With
Laser detection method detects the leucocyte blood sample, uses measurement angle for the big of 1 ~ 6 ° of first angle scattered light measuring cell
Small information, and use measurement angle for the grown form information of 6 ~ 18 ° of second angle scattered light measuring cell, it is thin with what is measured
The size information of born of the same parents and the grown form information of cell draw the first-second angle scatter plot, and referred to as main scatter plot dissipates the master
Point diagram carries out clustering, and the preliminary classification result of the eosinophil of acquisition is as shown in the a-quadrant in Fig. 3, it is clear that due to
Shadow cell extends to the right to be overlapped with eosinophil position, so that shadow cell and acidophil are mixed in together, influences acidophilus
The classification results of property granulocyte.
For above situation, the present embodiment is obtained by using the third angle scattered light signal that measurement angle is 26 ~ 42 °
The assisted morphometric information of cell, and formed with the assisted morphometric information of the cell and the grown form information of the above-mentioned cell measured
Second-third angle scatter plot, referred to as auxiliary scatter plot；Clustering is carried out to the auxiliary scatter plot, it is thin to obtain acidophil granules
The correcting region of born of the same parents；For eosinophil, third angle scattered signal and second angle scattered signal intensity are suitable, because
And the correcting region of eosinophil is located at the right regions of auxiliary scatter plot, as shown in the B area in Fig. 4；Then it will lead
It is classified as eosinophil on scatter plot but does not fall within the point in B area rejecting from eosinophil, remaining point is
The true classification results of eosinophil, as shown in the region C in Fig. 5.
Embodiment 2
At room temperature, the red blood cell in another whole blood sample is completely dissolved using hemolytic agent, obtains another leucocyte blood sample
This；A kind of leucocyte blood sample is detected with laser detection method, uses and measures angle as 1 ~ 6 ° of first angle scattered light measuring
The size information of cell, and use measurement angle for the grown form information of 6 ~ 18 ° of second angle scattered light measuring cell, with
The size information of the cell measured and grown form information the first-second angle scatter plot of drafting of cell, referred to as main scatter plot,
The main scatter plot of the leucocyte blood sample is as shown in fig. 6, from fig. 6, it can be seen that using general clustering method to this
Leucocyte blood sample is classified, and cannot obtain effective classification information.
For above situation, projective transformation can be carried out to dialogue cellular blood sample to by cervical arthroplasty or main scatter plot
This progress preliminary classification；The present embodiment is to carry out preliminary classification result by carrying out projective transformation to main scatter plot.Specifically, right
Point on above-mentioned main scatter plot carries out projective transformation to X direction and y direction respectively, obtains two projection histograms, so
The disposal of gentle filter is carried out using gaussian filtering method to projection histogram afterwards, obtains smooth histogram, as shown in Figure 7；It is smooth straight
There are two wave crests in square figure, and the threshold value using the wave trough position between wave crest as segmentation carries out preliminary classification to main scatter plot, obtain
To the leucocyte blood sample in monocyte preliminary classification result as shown in the region D in Fig. 8, it is clear that since monokaryon is thin
Boundary is unclear between born of the same parents and neutrophil leucocyte, and the preliminary classification result for obtaining monocyte is inaccurate.
For this purpose, the third angle scattered light signal that the present embodiment is 26 ~ 42 ° by using measurement angle, obtains cell
Assisted morphometric information, and second-is formed with the assisted morphometric information of the cell and the grown form information of the above-mentioned cell measured
Third angle scatter plot, referred to as auxiliary scatter plot；Clustering is carried out to the auxiliary scatter plot, obtains the amendment area of monocyte
Domain, since the internal complexity of lymphocyte and monocyte is low, in the lower left corner of the auxiliary scatter plot, formed one it is compact
Zonule, as shown in the region E in Fig. 9, the cell in the region includes lymphocyte and monocyte；Then by main scatter plot
On be classified as monocyte but do not fall within the point in the region E rejecting from monocyte, remaining point is the true of monocyte
Classification results, as shown in the region F in Figure 10.
In conclusion the present invention provides a kind of classification processing method of leucocyte and its devices.Specifically, pass through increase
Third angle scattered light signal is collected, and second angle scattered light signal is the longitudinal axis, third angle scattered light signal is that horizontal axis is drawn
System auxiliary scatter plot, repairs the preliminary classification result of main scatter plot followed by the cluster analysis result of auxiliary scatter plot
Just, the accuracy of leukocyte differential count is improved；And the universality of the classification processing method of leucocyte of the invention is good.
It should be understood that the application of the present invention is not limited to the above for those of ordinary skills can
With improvement or transformation based on the above description, all these modifications and variations all should belong to the guarantor of appended claims of the present invention
Protect range.
Claims (9)
1. a kind of classification processing method of leucocyte, which is characterized in that comprising steps of
A, the red blood cell in whole blood sample is completely dissolved using reagent, obtains leucocyte blood sample；
B, by light scattering testing technology obtain leucocyte blood sample in each cell particle first angle scattered light signal,
Second angle scattered light signal and third angle scattered light signal；
It C, is that the longitudinal axis draws main scatter plot using second angle light scattering signal as horizontal axis, first angle scattered light signal, with third
Angle scattered light signal is horizontal axis, second angle scattered light signal is that the longitudinal axis draws auxiliary scatter plot；
D, preliminary classification is carried out to main scatter plot by clustering, judges whether preliminary classification information is effective；
Preliminary classification information is effective, thens follow the steps E；
Preliminary classification information is invalid, then carries out preliminary classification to main scatter plot by projective transformation, obtain effective preliminary classification
Information, then execute step E；
E, using the cluster analysis result of auxiliary scatter plot, the effective preliminary classification information is modified, is obtained white thin
The true classification information of born of the same parents.
2. the classification processing method of leucocyte according to claim 1, which is characterized in that in step A, the reagent is molten
Blood agent.
3. the classification processing method of leucocyte according to claim 1, which is characterized in that in step B, the first angle
Scattered light signal represents the information of cell size, and the second angle scattered light signal represents the information of cellular morphology, and described
Three angle scattered light signals represent the auxiliary information of cellular morphology.
4. the classification processing method of leucocyte according to claim 1, which is characterized in that in step B, the first angle
The corresponding angle of scattered light signal is 1 ~ 6 °；The corresponding angle of the second angle scattered light signal is 6 ~ 18 °；The third angle
Spending the corresponding angle of scattered light signal is 26 ~ 42 °.
5. the classification processing method of leucocyte according to claim 1, which is characterized in that it is described by projective transformation to master
Scatter plot carries out preliminary classification and specifically includes step:
D1, projective transformation is carried out to its X direction and y direction to the point on main scatter plot respectively, it is straight to obtain two projections
Fang Tu；
D2, the disposal of gentle filter then is carried out to projection histogram, obtains smooth histogram；
Main scatter plot is carried out preliminary classification by D3, the threshold value using the wave trough position of the smooth histogram as segmentation.
6. leukocyte differential count processing method according to claim 5, which is characterized in that it is described projection histogram is carried out it is flat
The method of sliding filtering processing is gaussian filtering method.
7. the classification processing method of leucocyte according to claim 1, which is characterized in that the step E is specifically included:
E1, clustering is carried out to the auxiliary scatter plot, obtains the correcting region of certain class cell in leucocyte；
E2, the point for the correcting region that preliminary classification in main scatter plot does not fall within certain class cell for certain described class cell is picked
It removes, remaining point is the true group minute of certain class cell.
8. the classification processing method of leucocyte according to claim 7, which is characterized in that certain described class cell is acidophilia
Granulocyte or monocyte.
9. a kind of device of the classification processing method of leucocyte as described in claim 1 characterized by comprising
Fluid driving unit, for driving leucocyte blood sample；
Flow pool unit, for drive the cell particle in leucocyte blood sample individually sequentially through；
Light illuminating unit, for irradiating the cell particle for sequentially flowing through flowing pool unit；
Optical measurement unit, for measuring and obtaining the scattered light signal of cell particle；
The optical measurement unit includes:
First diaphragm, for measuring the first angle scattered light signal of cell particle；
Second diaphragm, for measuring the second angle scattered light signal of cell particle；
Third diaphragm, for measuring the third angle scattered light signal of cell particle；
First optical signal receiver, for obtaining the first angle scattered light signal of cell particle；
Second optical signal receiver, for obtaining the second angle scattered light signal of cell particle；
Third optical signal receiver, for obtaining the third angle scattered light signal of cell particle.
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| CN114419619A (en) * | 2022-03-29 | 2022-04-29 | 北京小蝇科技有限责任公司 | Erythrocyte detection and classification method and device, computer storage medium and electronic equipment |
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