CN111912978A

CN111912978A - Method and device for classifying and counting white blood cells and blood analyzer

Info

Publication number: CN111912978A
Application number: CN201910386076.5A
Authority: CN
Inventors: 陈鹏震; 孔繁钢
Original assignee: Shenzhen Mindray Bio Medical Electronics Co Ltd
Current assignee: Shenzhen Mindray Animal Medical Technology Co Ltd
Priority date: 2019-05-09
Filing date: 2019-05-09
Publication date: 2020-11-10

Abstract

The invention provides a method and a device for classifying and counting white blood cells and a blood analyzer, wherein the method comprises the following steps: obtaining an initial leukocyte histogram of leukocytes in a blood sample, wherein the initial leukocyte histogram is a histogram of the number and volume distribution of leukocytes; delineating a ghost region in the initial white blood cell histogram; correcting the ghost area to obtain a corrected lymphocyte area. According to the method, the device and the blood analyzer for counting and classifying the white blood cells, the interference of a ghost area on a lymphocyte area in a white blood cell histogram can be weakened or even eliminated, and the accuracy of counting and classifying the white blood cells is improved.

Description

Method and device for classifying and counting white blood cells and blood analyzer

Technical Field

The present invention relates generally to the field of blood analysis technology, and more particularly, to a method and apparatus for classifying and counting leukocytes, and a blood analyzer.

Background

Impedance method is one of the classic methods for counting and classifying human body white blood cells, and has wide application in counting and classifying animal white blood cells. The white blood cell count and classification of animals play an important role in the diagnosis and treatment of animals. On the other hand, because human blood cells and animal blood cells have certain differences in morphology, reaction conditions, reaction temperature and reagent dosage are not completely the same in the actual detection process; particularly, the using amount of the hemolytic agent has a remarkable influence on the detection of the animal blood cells. For example: the hemolytic agent with the same quantity and concentration can destroy red blood cells of a human, the white blood cell histogram of human blood has better morphology and can completely meet the requirements of counting and classification, but the red blood cells of some animals cannot be destroyed, so that a large amount of red blood cell fragments exist in a lymphocyte area at the front section of the white blood cell histogram of the animal, and the classification and counting of the white blood cells of the animal are interfered; if the dosage of the hemolytic agent is increased, the damage to the animal white blood cells is more obvious, so that the volume distribution of each particle swarm in a white blood cell histogram is more overlapped, and the classification result is seriously inaccurate. Therefore, in the current animal leukocyte detection, the lymphocyte region of the leukocyte histogram is inevitably interfered by cell debris, and the counting and classification of the leukocyte are influenced; this is a challenge in the detection of animal leukocytes by the current impedance method.

Disclosure of Invention

The present invention has been made to solve at least one of the above problems. Specifically, one aspect of the present invention provides a method for differential counting of leukocytes, the method comprising:

obtaining an initial leukocyte histogram of leukocytes in a blood sample, wherein the initial leukocyte histogram is a histogram of the number and volume distribution of leukocytes;

delineating a ghost region in the initial white blood cell histogram;

correcting the ghost area to obtain a corrected lymphocyte area.

Illustratively, delineating a ghost region in the initial white blood cell histogram includes:

acquiring a first boundary line of a ghost area and a lymphocyte area in the initial leukocyte histogram, wherein the ghost area is an area with a volume smaller than the volume corresponding to the first boundary line.

Illustratively, a method of delineating a ghost region in the initial white blood cell histogram, comprising:

demarcating a region of the initial leukocyte histogram having a volume less than a critical volume as the ghost region, the first boundary line being located at the critical volume.

Illustratively, obtaining a first boundary line of a ghost region and a lymphocyte region in the initial white blood cell histogram comprises:

obtaining a first peak point of the initial leukocyte histogram along the direction of volume increase starting from the minimum volume;

determining the first boundary line based on the first peak point.

Illustratively, the determining the first boundary line based on the first peak point includes:

the first boundary line is disposed at a position spaced apart from the volume of the first peak point by a predetermined volume, and the volume corresponding to the first boundary line is larger than the volume of the first peak point.

and searching a critical point with the slope being greater than the threshold slope for the first time in the initial leukocyte histogram curve from the first peak point along the direction of volume increase, wherein the first boundary line is a straight line passing through the critical point and perpendicular to the horizontal axis of the coordinate system.

Illustratively, the threshold slope is less than or equal to zero.

obtaining a first valley point of the initial white blood cell histogram from a minimum volume along a direction of volume increase;

determining the first boundary line based on the first valley point, wherein the first boundary line is disposed at a position spaced apart from a volume of the first valley point by a predetermined volume, and the volume corresponding to the first boundary line is smaller than the volume of the first valley point.

searching a first peak point on the initial leukocyte histogram from the maximum volume in the direction of volume reduction;

the first boundary line is disposed at a position spaced apart from the first peak point by a predetermined volume, and a volume corresponding to the first boundary line is smaller than a volume of the first peak point.

based on the results of at least two white blood cell counts, a ghost region is delineated in the initial white blood cell histogram.

Illustratively, the two white blood cell counts include a first white blood cell count and a second white blood cell count, the histogram of the first white blood cell count is the initial white blood cell histogram, and a shadow region is defined in the initial white blood cell histogram according to the results of the at least two white blood cell counts, including:

obtaining a threshold area of the ghost region based on an area of the histogram of the first white blood cell count and a threshold proportion coefficient of the ghost region in the histogram of the first white blood cell count, wherein the threshold proportion coefficient is obtained based on a result of the first white blood cell count and a result of the second white blood cell count;

determining a first boundary line of a ghost region and a lymphocyte region in the initial leukocyte histogram based on the threshold area to delineate a ghost region in the initial leukocyte histogram, wherein the first boundary line is located where an area in the histogram of the first leukocyte count is greater than or equal to the threshold area.

Illustratively, the amount of hemolytic agent used for the first white blood cell count is less than the amount of hemolytic agent used for the second white blood cell count.

Illustratively, the threshold area SGhost of the ghost region satisfies the following equation:

S_Ghost/(S_Ghost+S_Wbc)＝(WBC1-WBC2)/WBC1

wherein S is_Ghost+S_WbcThe area of the histogram of the first white blood cell count, the resultant WBC1 of the first white blood cell count, the resultant WBC2 of the second white blood cell count, and the threshold scaling factor is the ratio of the difference between the resultant WBC1 of the first white blood cell count and the resultant WBC2 of the second white blood cell count to the resultant WBC1 of the first white blood cell count.

Illustratively, modifying the ghost region to obtain a modified lymphocyte region comprises:

and removing the ghost area, and compensating the lymphocyte area to obtain a corrected lymphocyte area.

Illustratively, the compensating the lymphocyte region comprises:

defining a compensation line for the lymphocyte region in a blank region of the initial leukocyte histogram excluding the shadow region to compensate for the lymphocyte region.

Illustratively, the compensation line passes through a predetermined point and a critical point located in the blank area, the critical point being an intersection of a first boundary line between the lymphocyte region and the ghost region and a curve of the initial leukocyte histogram.

Illustratively, the predetermined point is a point on the horizontal axis of a rectangular coordinate system in which the initial white blood cell histogram is located.

Illustratively, the compensation line includes at least one of a straight line and a curved line.

Illustratively, the compensation line is obtained based on a predetermined function equation, the method further comprising:

calculating a lymphocyte compensation particle number from each volume between the predetermined point and the critical point on the compensation line based on the function equation;

compensating the lymphocyte region based on the summed number of lymphocyte compensation particles for each volume.

Illustratively, the function equation includes at least one of a first order function, a second order function, a polynomial function of a third order and more than a third order, an exponential function, and a logarithmic function.

Illustratively, the quadratic function is a convex function or a concave function.

Illustratively, calculating a lymphocyte compensation particle number from each volume between the predetermined point and the critical point on the compensation line based on the function equation specifically includes:

calculating a lymphocyte compensation particle number from each integral volume between the predetermined point and the critical point on the compensation line based on the function equation, wherein the lymphocyte compensation particle number is an integer.

Illustratively, the method further comprises:

pre-classifying the white blood cells based on the initial white blood cell histogram to obtain different types of white blood cell regions before correcting the ghost regions to obtain corrected lymphocyte regions, wherein the different types of white blood cell regions comprise lymphocyte regions, and the pre-classification is at least two classifications; after correcting the ghost area to obtain a corrected lymphocyte area, a corrected leukocyte histogram is obtained based on the corrected lymphocyte area.

Illustratively, the method further comprises:

obtaining a corrected leukocyte histogram based on the corrected lymphocyte region;

classifying the white blood cells based on the corrected white blood cell histogram to obtain different types of white blood cell regions, wherein the different types of white blood cell regions comprise lymphocyte regions, and the classification is at least two classifications.

Illustratively, the classifying the white blood cells based on the modified white blood cell histogram includes:

acquiring a first valley point in the modified white blood cell histogram curve from the minimum volume along the direction of volume increase;

defining a second boundary line between the modified lymphocyte region and another leukocyte region having a larger volume than the lymphocyte based on the first valley point.

Illustratively, the first valley point is a valley point between a first peak point and a second peak point taken from a minimum volume in a direction in which the volume increases.

Illustratively, the second demarcation line is spaced from the first valley point by a first predetermined volume and corresponds to a volume less than the volume of the first valley point.

Illustratively, the other white blood cells include a first white blood cell, a second white blood cell and a third white blood cell, wherein the second boundary line is a boundary line between the lymphocyte and the first white blood cell, and the classifying the white blood cells based on the corrected histogram of white blood cells further includes:

demarcating a third boundary line between the first type of white blood cell and the second type of white blood cell based on the first valley point and demarcating a fourth boundary line between the second type of white blood cell and the third type of white blood cell based on the first valley point or the second peak point.

Illustratively, the third demarcation line is spaced from the first valley point by a second predetermined volume, and the fourth demarcation line is spaced from the first valley point by a third predetermined volume, the third predetermined volume being greater than the second predetermined volume.

Illustratively, the third demarcation line corresponds to a volume greater than the volume of the first valley point and the fourth demarcation line corresponds to a volume greater than the volume of the first valley point.

Illustratively, before the step of classifying the white blood cells based on the corrected white blood cell histogram, the method further comprises:

and carrying out filtering processing on the corrected leukocyte histogram.

Illustratively, the orthogonal coordinate system in which the initial white blood cell histogram and the corrected white blood cell histogram are located has a cell volume size as an abscissa and a cell particle number of different volumes as an ordinate, wherein the method further comprises:

and calculating the total number of the cell particles included in the different types of leukocyte areas based on the sum of the number of the cell particles corresponding to each volume in the different types of leukocyte areas in the corrected leukocyte histogram.

Illustratively, the initial white blood cell histogram is obtained based on electrical impedance measurements of the blood sample.

Illustratively, the white blood cells include animal white blood cells.

In another aspect, the present invention provides a device for differential counting of leukocytes, the device comprising one or more processors, working together or separately, for performing the aforementioned methods of counting and classifying leukocytes.

Illustratively, the apparatus further comprises:

a detecting device for detecting the white blood cells in the blood sample and outputting a pulse signal when the white blood cells pass through a detecting hole in the detecting device, wherein the number of the pulse signal is in direct proportion to the number of the cells, and the height of the pulse signal is in direct proportion to the volume of the cells;

and the counting device is used for acquiring the pulse signals and obtaining an initial leukocyte histogram based on the pulse signals.

In a further aspect, the present invention provides a blood analyzer, which is characterized in that the blood analyzer comprises the above-mentioned white blood cell differential counting device.

The method, the device and the blood analyzer for counting and classifying the white blood cells are characterized in that an initial white blood cell histogram of the white blood cells in a blood sample is obtained, wherein the initial white blood cell histogram is a histogram of the number and volume distribution of the white blood cells; delineating a ghost region in the initial white blood cell histogram; and correcting the ghost area to obtain a corrected lymphocyte area, so that the interference of the ghost area on the lymphocyte area in the leukocyte histogram is weakened or even eliminated, and the accuracy of counting and classifying leukocytes is improved.

Drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without inventive labor.

FIG. 1 shows a conventional human blood leukocyte triage histogram;

FIG. 2 shows a conventional dog blood-like leukocyte four-classification histogram;

FIG. 3 shows a white blood cell histogram obtained after increasing the amount and concentration of hemolytic agent in the same dog blood as shown in FIG. 2;

FIG. 4 shows a flow chart of a method of differential counting of leukocytes in an embodiment of the invention;

FIG. 5 shows a schematic diagram of a ghost region delineated in an initial white blood cell histogram in a first example of the present invention;

FIG. 6 shows a schematic representation of a ghost region delineated in an initial white blood cell histogram in a second example of the present invention;

FIG. 7 shows a schematic diagram of a ghost region delineated in an initial white blood cell histogram in a third example of the present invention;

FIG. 8 shows a schematic representation of a silhouette region demarcated in an initial white blood cell histogram in a fourth example of the present invention;

FIG. 9 shows a schematic representation of a silhouette region demarcated in an initial white blood cell histogram in a fifth example of the present invention;

fig. 10 shows a schematic diagram of a ghost region delineated in an initial white blood cell histogram in a sixth example of the present invention;

FIG. 11 shows a schematic diagram of compensation of lymphocyte particles on an initial histogram in an embodiment of the invention;

FIG. 12 shows a schematic diagram of compensation of lymphocyte particles on an initial histogram in another embodiment of the present invention;

FIG. 13 shows a schematic diagram of compensation of lymphocyte particles on an initial histogram in a further embodiment of the invention;

FIG. 14 is a diagram illustrating four classifications of a modified white blood cell histogram in accordance with one embodiment of the present invention;

FIG. 15 shows a schematic block diagram of an example electronic device for implementing the method and apparatus for differential white blood cell counting according to embodiments of the present invention;

fig. 16 shows a schematic block diagram of an apparatus for differential counting of leukocytes in an embodiment of the invention.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, exemplary embodiments according to the present invention will be described in detail below with reference to the accompanying drawings. It is to be understood that the described embodiments are merely a subset of embodiments of the invention and not all embodiments of the invention, with the understanding that the invention is not limited to the example embodiments described herein. All other embodiments, which can be derived by a person skilled in the art from the embodiments of the invention described herein without inventive step, shall fall within the scope of protection of the invention.

In the following description, numerous specific details are set forth in order to provide a more thorough understanding of the present invention. It will be apparent, however, to one skilled in the art, that the present invention may be practiced without one or more of these specific details. In other instances, well-known features have not been described in order to avoid obscuring the invention. It is to be understood that the present invention may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.

The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms "comprises" and/or "comprising," when used in this specification, specify the presence of stated features, integers, steps, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and/or groups thereof. As used herein, the term "and/or" includes any and all combinations of the associated listed items.

In the following, the white blood cell histogram of each type of sample shows the problems in the classification of animal white blood cells. Under the condition of the same amount and concentration of hemolytic agent, the white blood cell histogram of human sample is shown in figure 1, and the white blood cell histogram of animal (dog blood sample is taken as an example) is shown in figure 2; the white blood cell histogram of the blood sample from the same animal after further addition of hemolytic agent is shown in FIG. 3. In FIG. 1, GOHST is a blood shadow, LYM is a Lymphocyte (Lymphocyte), MID is a Neutrophil (MID-sized Cell), GRAN is a Neutrophil (Granulocyte), and in FIGS. 2 and 3, MON is a Monocyte (Monocyte), EOS is an Eosinophil (Eosinophil), NEU is a Neutrophil (Neutrophil).

As can be seen from the histogram shown in fig. 1, under the condition, the leukocyte histograms of the human blood samples have more distributed leukocyte clusters, and the interference of the lymphocyte regions by the ghost regions is less, so that a good leukocyte counting and classifying effect can be achieved; however, as shown in fig. 2, although the distribution of each leukocyte population is relatively open, LYM particles in the histogram are relatively severely interfered by GHOST, which significantly affects the accuracy of LYM classification; when increasing hemolytic agent, the white blood cell histogram of the animal (e.g. dog) is shown in fig. 3, although the effect of LYM by GHOST is greatly reduced, the particle size of the whole white blood cell histogram is more tightly compressed, which is not favorable for white blood cell classification (especially LYM, MON).

Therefore, in order to solve the technical problem that during the detection of a blood sample (especially an animal blood sample), a lymphocyte region in a leukocyte histogram is interfered by ghosts (i.e. cell debris), so that the leukocyte count and classification are inaccurate, the embodiment of the invention provides an improved method for counting leukocytes by classification, which comprises the following steps: obtaining an initial leukocyte histogram of leukocytes in a blood sample, wherein the initial leukocyte histogram is a histogram of the number and volume distribution of leukocytes; delineating a ghost region in the initial white blood cell histogram; correcting the ghost area to obtain a corrected lymphocyte area. The method weakens or even eliminates the interference of the ghost area on the lymphocyte area in the leukocyte histogram, improves the accuracy of lymphocyte classification and counting, and further improves the accuracy of leukocyte counting and classification.

In order to provide a thorough understanding of the present invention, a detailed structure will be set forth in the following description in order to explain the present invention. Alternative embodiments of the invention are described in detail below, however, the invention may be practiced in other embodiments that depart from these specific details.

Specifically, the method for differential white blood cell count according to the present application will be described in detail below with reference to the accompanying drawings. The features of the following examples and embodiments may be combined with each other without conflict.

In one embodiment, as shown in fig. 4, the method for classifying and counting leukocytes in the embodiment of the present invention includes the following steps S401 to S403:

first, in step S401, an initial leukocyte histogram of leukocytes in a blood sample is obtained, wherein the initial leukocyte histogram is a histogram of the number and volume distribution of leukocytes.

The blood sample may be any animal or human blood sample including leukocytes, and may be obtained by treating whole blood drawn from an animal body with a diluent, a hemolytic agent, or the like. The diluent is an isotonic solution with acid-base buffering effect, proper ionic strength and conductivity, for example, the diluent mainly contains hypoxanthine or xanthine compound or its salt as main component or can be other diluents capable of playing the role, and the hemolytic agent is used for lysing red blood cells to classify and count white blood cells. The hemolytic agent includes a surfactant, which may specifically include a cationic surfactant and a nonionic surfactant. The amount and concentration of the hemolytic agent may be selected reasonably according to actual sample preparation requirements, and are not specifically limited herein. The hemolytic agent may be a quaternary ammonium salt ionic surfactant as a main component, or may be any other surfactant capable of exerting the above-described effects.

The initial histogram of white blood cells is a histogram of the number and volume distribution of white blood cells, for example, the histogram shown in the figure is represented by a rectangular coordinate system with the size of the cell volume as the abscissa and the number of cell particles (including white blood cells and ghosts, for example) of different volumes as the ordinate.

The initial histogram may be obtained by testing the blood sample by any suitable testing method, for example, the initial leukocyte histogram is obtained by testing the blood sample based on electrical impedance. Alternatively, the initial white blood cell histogram may be the animal white blood cell histogram shown in fig. 2.

With continued reference to fig. 4, in step S402, a ghost region is delineated in the initial white blood cell histogram.

The shadow region may be delineated in the initial leukocyte histogram by any suitable method, for example, by: obtaining a first boundary line of a ghost area and a lymphocyte area in the initial leukocyte histogram, wherein the ghost area is an area with a volume smaller than the volume corresponding to the first boundary line, and a ghost is a cell fragment generated after the erythrocyte is acted by the hemolytic agent, and generally has a volume smaller than that of the leukocyte, so that the ghost area is positioned on the left side of the first boundary line on the histogram, namely the ghost area is an area with a volume smaller than the volume corresponding to the first boundary line in the initial histogram. Optionally, the first boundary line is a straight line perpendicular to the abscissa axis in a rectangular coordinate system in which the initial histogram is located.

The ghost regions can be delineated in the initial histogram by a number of suitable methods, and the methods in the first through sixth examples are described in detail below with reference to fig. 5 through 10, but it should be understood that the following methods are merely examples and that other suitable methods may be applied to the present application.

In a first example, as shown in fig. 5, a method of delineating a ghost region in the initial white blood cell histogram, comprising: demarcating a region of the initial leukocyte histogram having a volume less than a critical volume as the ghost region, the first boundary line being located at the critical volume. The critical volume can be set reasonably according to a priori experience, for example, at specific reaction conditions, reaction temperature, reagent (including hemolytic agent and diluent) dosage, and through multiple tests, the critical volume under the specific conditions, especially specific hemolytic agent dosage, can be the position where the first boundary line of the ghost area and lymphocyte area in the initial leukocyte histogram in the prior experience is located, for example, as shown in fig. 5, the critical volume is 48fL, and the first boundary line is located at the critical volume. The critical volume may vary depending on the reaction conditions, reaction temperature, and reagent (including hemolytic agent and diluent) dosage, and may be adjusted according to the actual situation, for example, the critical volume may decrease with the increase of the amount and concentration of hemolytic agent.

Specifically, a first boundary line of a ghost area and a lymphocyte area in the initial leukocyte histogram may be obtained, where the ghost area is an area having a volume smaller than a volume corresponding to the first boundary line, that is, an area on the left side of the first boundary line in the initial histogram, and the first boundary line may be obtained by any suitable method, for example, a first peak point of the initial leukocyte histogram is obtained from a minimum volume along a direction of volume increase; the first boundary line is determined based on the first peak point, wherein the first boundary line is usually located at the left side of the first peak point, that is, the first boundary line corresponds to a volume larger than the volume of the first peak point, and the specific position of the first boundary line can be determined based on the first peak point by the following examples as shown in fig. 6 and 7.

In a second example as shown in fig. 6, said determining said first boundary line based on said first peak point comprises: and searching a critical point B with the slope being larger than a threshold slope K for the first time in the initial leukocyte histogram curve from the first peak point A along the direction of the volume increase, wherein the first boundary line is a straight line passing through the critical point B and perpendicular to the horizontal axis (also called the abscissa axis) of the rectangular coordinate system where the histogram is located. In general, the curve of the initial leukocyte histogram on the right side of the first peak point a has a downward trend, and the slope of a point on the curve of the initial leukocyte histogram in a predetermined segment from the first peak point a is less than or equal to 0, so the value of the threshold slope K is set to be less than or equal to zero, and specifically, the value of the threshold slope K may be set according to actual conditions. The area of the ghost can be determined directly from the critical point B, if the volume of the critical point B is equal to n (fL), the area of the initial leukocyte histogram where the volume V is smaller than n (fL) is the area of the ghost, for example, the threshold slope K is set to-2.6, the slope of the curve at the critical point B is-2.0, -2.0 is greater than 2.6, and the volume of the critical point B n (fL) is equal to 46fL, the area of the initial leukocyte histogram where the volume V is smaller than 46fL is the area of the ghost.

In a third example as shown in fig. 7, determining the first boundary line based on the first peak point includes: the first boundary line is arranged at a position separated from the volume of the first peak point by a predetermined volume M, and the volume corresponding to the first boundary line is larger than the volume of the first peak point, that is, the first boundary line is arranged at a position at the right side of the first peak point and separated from the volume of the first peak point by the predetermined volume M, the predetermined volume M can be reasonably set according to the prior experience, for example, under specific reaction conditions, reaction temperature, reagent (including hemolytic agent and diluent) dosage, the volume of the first peak point under the specific conditions, especially specific hemolytic agent dosage and the volume separated between the position of the first boundary line of the blood shadow region and the lymphocyte region in the initial leukocyte histogram and the first peak point are obtained through multiple detections, so as to determine the predetermined volume M, for example, as shown in fig. 7, different reaction conditions, reaction temperatures, reagent (including hemolytic agent and diluent) dosage, first peak position, and predetermined volume M may also be different, and may be adjusted according to practical situations, for example, the first peak position may move to the left with increasing amount and concentration of hemolytic agent, that is, the corresponding volume may decrease, and the predetermined volume M may also change, for example, may also decrease.

In a fourth example as shown in fig. 8, acquiring a first boundary line of a ghost region and a lymphocyte region in the initial leukocyte histogram, comprises: acquiring a first valley point N of the initial leukocyte histogram along the direction of volume increase starting from the minimum volume; determining the first boundary line based on the first valley point, wherein the first boundary line is disposed at a position spaced apart from a volume of the first valley point by a predetermined volume, and the volume corresponding to the first boundary line is smaller than the volume of the first valley point. A reasonable predetermined volume may be set a priori, for example if the first valley point has a volume of 70fL and the predetermined volume is 22fL, the first boundary line is located at 48fL, so that the ghost region is a region with a volume of less than 48fL on the initial histogram.

By way of further example, a shadow region may be defined in the initial leukocyte histogram by the method in the fifth example shown in fig. 9, specifically including: searching a first peak point P (e.g., a peak point of a Neu region) on the initial leukocyte histogram from the maximum volume in a direction of volume reduction (e.g., from the right side to the left side of the initial leukocyte histogram); the first boundary line Q is set at a position spaced apart from the first peak point P by a predetermined volume S and the volume corresponding to the first boundary line Q is smaller than the volume of the first peak point P, the first boundary line is located at a volume obtained by subtracting the volume of the first peak point from the predetermined volume. This predetermined volume can be set reasonably a priori, for example if the first peak point has a volume of 125fL, the predetermined volume S is 77fL, and the first boundary line is located at 48fL, so that the ghost region is a region with a volume less than 48fL on the initial histogram.

In other examples, a method of delineating a ghost region in the initial white blood cell histogram, comprising: based on the results of at least two white blood cell counts, a ghost region is delineated in the initial white blood cell histogram. Wherein the at least two white blood cell count results comprise two, three, or more white blood cell count results.

Optionally, the at least two white blood cell counts include a first white blood cell count and a second white blood cell count, the histogram of the first white blood cell count is the initial white blood cell histogram, and a shadow region is defined in the initial white blood cell histogram according to the results of the at least two white blood cell counts, including steps S1 and S2:

in step S1, obtaining a threshold area of the ghost region based on the area of the histogram of the first white blood cell count and a threshold proportion coefficient that the ghost region occupies in the histogram of the first white blood cell count; the above-mentioned threshold proportionality coefficient may be determined based on any suitable method, for example, the threshold proportionality coefficient is obtained based on the result of the first white blood cell count and the result of the second white blood cell count.

Illustratively, the amount of hemolytic agent used for the first white blood cell count is less than the amount of hemolytic agent used for the second white blood cell count, for example, assuming that white blood cells of an animal are counted twice, and a blood sample of the same animal, the histogram of the white blood cell classification obtained at the 1 st time may be shown in fig. 2, the result WBC1 of the first white blood cell count may be obtained by counting the histogram of the white blood cells, the result WBC2 of the second white blood cell count may be obtained by counting the histogram of the white blood cells obtained at the 2 nd time by increasing the amount and concentration of hemolytic agent as shown in fig. 3.

The result of the first white blood cell counting is higher due to the fact that different hemolysis doses contain ghost interference, the result of the second white blood cell counting is high due to the fact that the hemolysis agent is high, the influence of the ghost formed by cell fragments after red blood cells and platelets are damaged on the white blood cells is small and can be ignored, the result of the second white blood cell counting can be considered to be a true value, and WBC2 is not more than WBC 1; the threshold area SGhost of the ghost region satisfies the following equation:

S_Ghost/(S_Ghost+S_Wbc)＝(WBC1-WBC2)/WBC1

wherein S is_Ghost+S_WbcThe area of the entire histogram for the first white blood cell count, the resultant WBC1 for the first white blood cell count, the resultant WBC2 for the second white blood cell count, and the threshold scaling factor is the ratio of the difference between the resultant WBC1 for the first white blood cell count and the resultant WBC2 for the second white blood cell count to the resultant WBC1 for the first white blood cell count.

In another example, if a plurality of (e.g., greater than or equal to 3) animal white blood cell counts are performed; selecting a white blood cell histogram of one weak hemolysis (less hemolytic agent concentration and amount) as a second white blood cell count; the threshold area of the ghost area in the white blood cell histogram of the first white blood cell count can also be obtained based on the aforementioned method by selecting the white blood cell histogram of one count in other strong hemolysis doses (the hemolysis agent concentration and amount are larger, that is, the amount of the hemolysis agent is larger than the amount of the hemolysis agent used in the first white blood cell count) as the second white blood cell count.

In step S2, a first boundary line of the ghost region and the lymphocyte region in the initial leukocyte histogram is determined based on the threshold area to demarcate the ghost region in the initial leukocyte histogram, wherein the first boundary line is located at an area greater than or equal to the threshold area in the first leukocyte count histogram, for example, the first leukocyte count histogram sequentially overlaps the first boundary line with an area S starting from a minimum volume and increasing from the minimum volume to a volume, and the first boundary line of the first leukocyte count histogram is determined at a position where S is first greater than or equal to the threshold area, that is, the position of the first boundary line of the initial leukocyte histogram.

For example, in the sixth example shown in FIG. 10, the first WBC1 count is 18.4 10^9/L, the second WBC2 count is 13.6 ^ 10^9/L, the first WBC histogram is the entire area S_Ghost+S_Wbc13850; then S is calculated by the above formula_Ghost3613; by sequentially superimposing the areas S on the left side of the histogram of the first white blood cell count, with S being greater than or equal to S for the first time_GhostThe position is the first boundary line between the GHOST and the lymphocyte region, for example, the first boundary line is located at 49fL, and the GHOST region is located in the region of the leukocyte histogram with the first leukocyte count, which has a volume smaller than the volume corresponding to the first boundary line.

Subsequently, with continued reference to fig. 4, in step S403, the ghost region is corrected to obtain a corrected lymphocyte region.

After the positions of the ghost areas in the initial leukocyte histogram are determined based on the above steps, if the leukocyte classification, such as two-classification, three-classification, four-classification, or five-classification, is directly performed, the percentage of lymphocytes is low due to interference of the ghost on the lymphocyte areas, and therefore, the lymphocyte areas can be corrected by the method of this step.

In one specific example, modifying the ghost region to obtain a modified lymphocyte region comprises: and removing the ghost area, compensating the lymphocyte area, namely compensating lymphocyte particles on the initial histogram to obtain a corrected lymphocyte area, weakening or even eliminating the interference of the ghost area on the lymphocyte area in the leukocyte histogram, and improving the accuracy of counting and classifying leukocytes.

Fig. 3 shows that in the white blood cell histogram without ghost interference or with very little ghost interference, the lymphocyte particle cluster is generally a distribution region from about 20fL, and after the ghost region in the initial white blood cell histogram is removed, if the lymphocyte region is not compensated, the lymphocyte particle cluster is a distribution region from about 50fL, for example, and the 50fL is greater than 20fL, it can be seen that a part of the lymphocyte particles are also removed after the ghost is actually removed, which results in a low LYM classification and count, so the lymphocyte region of the white blood cell histogram after the ghost is removed needs to be compensated. The lymphocyte region can be compensated by any suitable method on the initial histogram, for example, a compensation line of the lymphocyte region is defined in a blank region of the initial leukocyte histogram after the ghost region is removed, so as to compensate the lymphocyte region, that is, the number of cell particles defined by the compensation line on the initial histogram is compensated to the number of particles of the lymphocyte population, and a reasonable number is defined by the compensation line to be compensated to the lymphocyte, so that the problem that the percentage of the lymphocyte region is low due to the interference of the ghost region on the lymphocyte region in the leukocyte histogram is solved, and the accuracy of counting and classifying the lymphocyte is improved.

The compensation line may include at least one of a straight line and a curved line, for example, a straight line, a curved line, or a line formed by connecting a plurality of straight lines and curved lines end to end, or a line formed by connecting at least two straight lines with different slopes end to end, or a line formed by connecting different curved lines end to end, or other suitable lines.

In one example, the compensation line passes through a predetermined point located in a blank area after the ghost area is removed and a critical point, the critical point being an intersection of a first boundary line between the lymphocyte area and the ghost area and a curve of the initial leukocyte histogram. The predetermined point can be chosen reasonably according to the actual need, for example, the predetermined compensation is to the number of particles in the lymphocyte population to set reasonably, or the predetermined point can be set a priori. Optionally, the predetermined point is a point on a horizontal axis of a rectangular coordinate system of the initial white blood cell histogram, that is, the number of cell particles at the coordinate of the predetermined point is zero.

In one example, the compensation line may be obtained based on a predetermined function equation including at least one of a first order function, a second order function, a polynomial function of three or more orders, an exponential function, a logarithmic function, or other suitable function equations, but it is worth mentioning that no matter which kind of function equation is based on the obtained compensation line, the compensation line must also satisfy the passing of a predetermined point and a critical point, or the predetermined function equation may be obtained by first setting the type of the predetermined function equation, such as a first order function or a second order function, and then using the coordinates of the predetermined point and the critical point in the initial histogram to obtain the predetermined function equation through calculation.

Further, the method further comprises: calculating a lymphocyte compensation particle number from each volume between the predetermined point and the critical point on the compensation line based on the function equation; the lymphocyte region is compensated based on the sum of the lymphocyte compensation particle numbers of each volume, that is, the particle number of the lymphocyte population is compensated, and preferably, the lymphocyte compensation particle numbers of each integral volume between the predetermined point and the critical point on the compensation line are calculated based on the function equation, wherein the lymphocyte compensation particle number is an integer. Alternatively, the lymphocyte compensation particle number may be obtained based on a product of a ratio of an area of a region defined by the compensation line in an area of the entire initial leukocyte histogram and a total cell particle number represented by the initial leukocyte histogram, and the lymphocyte compensation particle number is rounded in the calculation process.

Several methods of compensating for lymphocyte regions are described in detail below with reference to fig. 11-13, but it should be understood that the following methods are exemplary only and that other suitable methods are equally applicable to the present application.

In the example shown in fig. 11, the lymphocyte regions in the initial histogram are compensated using straight lines, for example using the equation of a straight line y k V + b, k > 0. First, the blood shadow region defined by the above steps is removed, and then a straight line is taken from a predetermined point a to a critical point B as a LYM compensation part on the left side of the initial leukocyte histogram, as shown in fig. 11, wherein the critical point B is an intersection point of a first boundary line between the determined GHOST and LYM and the initial leukocyte histogram curve, the predetermined point a can be reasonably set according to practical situations, for example, the coordinates of the predetermined point a in the initial leukocyte histogram are (20,0), and the coordinates of the critical point B can be determined according to the above steps in defining the blood shadow region, for example, an equation of an AB straight line can be calculated according to the coordinates of the predetermined point a and the critical point B (y is 5.8 (V-20), V is a cell particle volume between the point a and the point B, and y is a cell particle number with a volume of V), further, the number of Lymphocyte (LYM) compensation particles per volume (where the volumes are all integers) between the point a and the point B (for example, 50fL after the volume of the point B is rounded) can be calculated by the linear equation (the rounding operation is also performed on the number of particles calculated by the linear equation), and the number of particles of the lymphocyte population is compensated by the sum of the number of lymphocyte compensation particles per volume.

In the example shown in fig. 12 and 13, the LYM region is compensated using a quadratic polynomial (i.e., a quadratic function) of y ═ a × V²+ b V + c. The ghost area defined by the previous steps is removed, and then a quadratic polynomial compensation is made from a predetermined point A (which can be reasonably set according to practical situations, such as A (20,0)) to a critical point B to serve as a left LYM compensation part of the white blood cell histogram. This quadratic polynomial may be a convex or a concave function. If the convex function is illustrated in fig. 12, the AB point equation may be: y-0.48V²+ 30.72V-421.52; an example of a concave function is shown in fig. 13, and the AB point equation can be: y 0.48V²-19.2 x V + 192. The LYM compensated population for each volume size (where the volumes are all integers) between point a and point B (e.g., point B is rounded to 50fL) can also be calculated by the equation (the population calculated by the equation is also rounded), and the population of the lymphocyte population is compensated for by the sum of the compensated populations of lymphocytes for each volume size.

In one example, the method of an embodiment of the present invention further comprises: pre-classifying the white blood cells based on the initial white blood cell histogram to obtain different types of white blood cell regions before correcting the ghost regions to obtain corrected lymphocyte regions, wherein the different types of white blood cell regions comprise lymphocyte regions, and the pre-classification is at least two classifications; after correcting the ghost region to obtain a corrected lymphocyte region, a corrected leukocyte histogram is obtained based on the corrected lymphocyte region, the corrected leukocyte histogram being already a classified leukocyte histogram, and then the number of particles per classification can be statistically calculated based on the corrected leukocyte histogram.

In another example, the method of an embodiment of the present invention further comprises: obtaining a corrected leukocyte histogram based on the corrected lymphocyte region; classifying the leukocytes based on the corrected leukocyte histograms to obtain different leukocyte regions, wherein the different leukocyte regions include lymphocyte regions that are modified by the method of the previous step, and the classification is at least two classifications, such as two classifications, three classifications, four classifications, five classifications, and the like, wherein the classification of the leukocytes may be different according to the blood sample type, for example, in the case of animal leukocytes such as dogs, the leukocytes are classified into lymphocyte populations LYM, monocyte populations MON, neutrophil populations NEU, and eosinophil populations EOS in sequence from the smallest volume along the direction of volume increase.

Classifying the leukocytes in the modified leukocyte histogram can be performed by any suitable method, for example, in one example, classifying the leukocytes based on the modified leukocyte histogram includes: acquiring a first valley point in the modified leukocyte histogram curve from the minimum volume along the direction of volume increase, optionally, the first valley point is a valley point between a first peak point and a second peak point acquired from the minimum volume along the direction of volume increase; a second boundary line between the modified lymphocyte region and another leukocyte region having a larger volume than the lymphocyte is defined based on the first valley point, optionally, the second boundary line is separated from the first valley point by a first predetermined volume, and the volume corresponding to the second boundary line is smaller than the volume of the first valley point, that is, the second boundary line is located on the left side of the first valley point. The first predetermined volume can be reasonably set a priori, and can be properly adjusted for different detection conditions, and is not specifically limited herein.

Alternatively, the leukocytes include, in addition to the lymphocytes included therein, a first type of leukocytes, which are a monocyte population MON, a neutrophil population NEU, and an eosinophil population EOS, which are sequentially increased in volume, a second type of leukocytes, which are boundaries between the lymphocytes and the first type of leukocytes, for example, as shown in fig. 14. Further demarcating, by any suitable method, a third demarcation line between the first type of white blood cells and the second type of white blood cells, and a fourth demarcation line between the second type of white blood cells and the third type of white blood cells, for example, classifying the white blood cells based on the modified white blood cell histogram, further comprises: demarcating a third demarcation line between the first type of white blood cell and the second type of white blood cell and a fourth demarcation line between the second type of white blood cell and the third type of white blood cell based on the first valley point. Wherein the third demarcation line is spaced from the first valley point by a second predetermined volume, the fourth demarcation line is spaced from the first valley point by a third predetermined volume, and the third predetermined volume is greater than the second predetermined volume. The volume corresponding to the third boundary line is greater than the volume of the first valley point, and the volume corresponding to the fourth boundary line is greater than the volume of the first valley point, that is, the third boundary line and the fourth boundary line are both located on the right side of the first valley point, and the fourth boundary line is located on the right side of the third boundary line.

The aforementioned first, second and third predetermined volumes may be reasonably set according to a priori experience, for example, according to the result of the classification of the leukocyte histogram in the priori experience, for example, the spacing volume between the volume corresponding to each boundary line and the volume of the first valley point, so as to make the classification more accurate.

In one example, before the step of classifying the white blood cells based on the corrected white blood cell histogram, the method further includes: and a step of performing filtering processing on the corrected leukocyte histogram, wherein the filtering processing can remove noise in the corrected leukocyte histogram, so that the result of counting the corrected leukocyte histogram is more accurate.

The above classification method is also equally applicable to pre-classifying the initial leukocyte histogram.

For example, the method of using the convex function to perform LYM compensation in fig. 12 is used to obtain the modified leukocyte histogram shown in fig. 14, filter the modified leukocyte histogram, then find the first peak point a and the second peak point B in the modified leukocyte histogram, find the first valley point C between the AB points, respectively, the left and right certain distances (i.e., the predetermined intervals) of the first valley point C are the boundary between LYM and MON and the boundary between MON and NEU, respectively, and the right certain distance (i.e., the predetermined interval) of the second peak point B is found as the boundary between NEU and EOS, or the right certain distance (i.e., the predetermined interval) is found as the boundary between NEU and EOS based on the first valley point, thereby completing the fourth classification of the modified leukocyte histogram.

Animal samples (in the case of dog blood) were randomly selected and sorted by LYM compensation of leukoreduced histograms as shown in the following table:

it can be seen from the above table that the classification result of the corrected lymphocyte region obtained after compensation is performed on the lymphocytes based on the method of the present application is closer to the reference value (i.e., the true value) than that before compensation, so that the interference of the ghost region on the lymphocyte region in the leukocyte histogram can be weakened or even eliminated based on the method of the present application, the accuracy of classification and counting of the lymphocytes is improved, and the accuracy of counting and classifying of the leukocytes is further improved.

After the modified leukocyte histogram is classified, the number of cell particles included in each classification result may be counted based on the classification result, and in one example, the orthogonal coordinate system in which the initial leukocyte histogram and the modified leukocyte histogram are located uses the cell volume as an abscissa, and the total number of cell particles included in different types of leukocyte regions including a region defined based on the above-mentioned compensation line is calculated based on a sum of numbers (or integers) of cell particles corresponding to each volume (the volume may be rounded) in the different types of leukocyte regions in the modified leukocyte histogram. In other examples, the total number of cell particles of each cell type may be calculated based on the area occupied by each cell type region in the corrected leukocyte histogram, for example, the total number of cell particles may be obtained based on the initial leukocyte histogram, the total number of particles of the lymphocyte type corresponding to the corrected lymphocyte type may be calculated based on the percentage of the area of the corrected lymphocyte type region in the total area of the initial leukocyte histogram by multiplying the total number of cell particles by the percentage, and the number of particles of other leukocyte classes may be calculated according to the same method.

In summary, according to the method of the embodiment of the present invention, an initial leukocyte histogram of leukocytes in a blood sample is obtained, wherein the initial leukocyte histogram is a histogram of the number and volume distribution of leukocytes; delineating a ghost region in the initial white blood cell histogram; and correcting the ghost area to obtain a corrected lymphocyte area, so that the interference of the ghost area on the lymphocyte area in the leukocyte histogram is weakened or even eliminated, and the accuracy of counting and classifying leukocytes is improved.

An example electronic device 150 for implementing the method and apparatus for differential white blood cell counting of embodiments of the present invention is described below with reference to fig. 15.

In one example, as shown in fig. 15, electronic device 150 may include one or more processors 151, one or more storage devices 152, an input device 153, an output device 154, a communication interface 155, interconnected by a bus system 156 and/or other form of connection mechanism (not shown). It should be noted that the components and configuration of the electronic device 150 shown in FIG. 15 are exemplary only, and not limiting, and the electronic device may have other components and configurations as desired.

The processor 151 may be a Central Processing Unit (CPU), a Graphics Processing Unit (GPU), an Application Specific Integrated Circuit (ASIC), a Field Programmable Gate Array (FPGA) or other form of processing unit having data processing capabilities and/or instruction execution capabilities, and may control other components in the apparatus for white blood cell count classification to perform the desired functions. The processor is capable of executing programs and/or instructions stored in the storage device to perform the methods of white blood cell count classification described herein. For example, processor 151 can include one or more embedded processors, processor cores, microprocessors, logic circuits, hardware Finite State Machines (FSMs), Digital Signal Processors (DSPs), or a combination thereof.

The storage 152 may include one or more computer program products that may include various forms of computer-readable storage media, such as volatile memory and/or non-volatile memory. The volatile memory may include, for example, Random Access Memory (RAM), cache memory (cache), and/or the like. The non-volatile memory may include, for example, Read Only Memory (ROM), hard disk, flash memory, etc. On which one or more computer program instructions may be stored that may be executed by processor 151 to implement the functions of the embodiments of the invention described herein (as implemented by the processor) and/or other desired functions. Various applications and various data, such as various data used and/or generated by the applications, may also be stored in the computer-readable storage medium.

The input device 153 may be a device used by a user to input an instruction (for example, in the method for classifying a white blood cell count according to the embodiment of the present invention described herein, the user may input a function equation related to the compensation line, a predetermined volume when dividing the boundary line, and the like), and may include one or more of a keyboard, a mouse, a microphone, a touch screen, and the like. In addition, the input device 153 may be any interface for receiving information.

The output device 154 may output various information (e.g., images or sounds) to the outside (e.g., a user), and may include one or more of a display (e.g., a list of parameters requiring the white blood cell count classification device, a result of the white blood cell count classification, a histogram of the white blood cell classification, etc., displayed to the user), a speaker, and the like.

The communication interface 155 is used for communication between the electronic device 150 and other devices, including wired or wireless communication. The electronic device 150 may access a wireless network based on a communication standard, such as WiFi, 2G, 3G, 4G, 5G, or a combination thereof. In an exemplary embodiment, the communication interface 155 further includes a Near Field Communication (NFC) module to facilitate short-range communications. For example, the NFC module may be implemented based on Radio Frequency Identification (RFID) technology, infrared data association (IrDA) technology, Ultra Wideband (UWB) technology, Bluetooth (BT) technology, and other technologies.

Exemplarily, an exemplary electronic device for implementing the method of white blood cell count classification according to an embodiment of the present invention may be implemented as a terminal such as a desktop computer, a tablet computer, or the like, or an apparatus for white blood cell count classification and a blood analyzer including the terminal, or the like.

The device for differential counting of leukocytes provided by another aspect of the invention is described below with reference to fig. 16. Fig. 16 shows a schematic block diagram of an apparatus for differential counting of leukocytes in an embodiment of the invention.

As shown in fig. 16, the apparatus 160 for white blood cell count classification further comprises a reaction device 162 and a sampling device 161, wherein the reaction device 162 is used for preparing a blood sample to be tested into a blood sample, and for example, the reaction device comprises a reaction cell. The sampling device 161 is used for injecting a blood sample to be tested into a reaction device such as a reaction cell, for example, for injecting whole blood of a drawn animal sample into the reaction cell, and in this embodiment, the sampling device 161 is a sampling needle. In other embodiments, the sampling device may be implemented in other ways.

In one example, as shown in fig. 16, the apparatus 160 for white blood cell count sorting further includes a reagent storage device 163, and the reagent storage device 163 is connected to the reaction device 162 for supplying a reagent for preparing a blood sample, such as a hemolytic agent, a diluent, etc., into the reaction device 162. The number of the reagent storage devices is set according to the kind of the reagent, and for example, the reagent storage device includes a storage device for storing a hemolytic agent and a storage device for storing a diluent. After the reagent is injected into the reaction device, a blood sample, a hemolytic agent, a diluent, and the like are mixed in the reaction device to prepare a blood sample for detecting leukocytes.

The apparatus 160 for white blood cell count classification further comprises a conveying device (not shown) for conveying the sample liquid in the reaction cell to the detection device, in this embodiment, the conveying device comprises an injector and a conveying pipeline communicated with the injector, the sample liquid outlet of the reaction cell and the inlet of the detection device are communicated through the conveying pipeline, the number of the injectors can be multiple, and each injector performs suction and discharge actions under the control of the control device.

In one example, as shown in fig. 16, the device 160 further comprises a detecting device 164, wherein the detecting device 164 is used for detecting white blood cells in the blood sample and outputting pulse signals when the white blood cells pass through a detecting hole in the detecting device, the number of the pulse signals is proportional to the number of the cells, the height of the pulse signals is proportional to the volume of the cells, and thus the number and the volume of the blood cell particles in the blood are obtained. Wherein, the detection device is used for detecting the blood sample prepared by the reaction tank.

In one example, the structure of the detection device 164 may include a counting cell (not shown) and a pulse sensor (not shown), optionally, the pulse sensor may include a small-hole tube, the small-hole tube is provided with a detection small hole, optionally, the diameter of the detection small hole is less than 100 micrometers, the thickness of the detection small hole ranges from 60 micrometers to 90 micrometers, for example, about 75 micrometers, the counting cell is filled with a conductive solvent, and the detection small hole is divided into a front cell and a rear cell; the front pool and the back pool are respectively provided with a positive electrode and a negative electrode, the positive electrode and the negative electrode are connected with one end of a constant current source, the electrode arranged in the front pool, a conductive solvent, the electrode arranged in the back pool and the constant current source form a closed loop which is connected in series, after the power supply is switched on, the electrodes positioned at two sides of the small hole tube generate stable current, diluted cell suspension flows to the inside of the small hole tube from the outer side of the small hole tube through the detection small hole, the resistance in a small hole induction area is increased, instantaneous voltage change is caused to form a pulse signal, the amplitude of the pulse signal is in direct proportion to the size of the cell volume, the pulse number is in direct proportion to the cell number, the blood cell number and the volume value in a blood sample are obtained, and different types.

In one example, the apparatus 160 further comprises a counting device 165 for acquiring the pulse signal, and obtaining an initial white blood cell histogram based on the pulse signal. The counting device receives the pulse signal output by the detection device and analyzes the pulse signal, so that an initial leukocyte histogram is obtained based on the pulse signal. The counting device may also be used to count the initial leukocyte histogram statistically to obtain the result of the leukocyte count, such as obtaining the total number of cell particles in the entire initial leukocyte histogram. The counting means may be implemented on the basis of the aforementioned electronics, which may be integrated in the device 160 or may be separate from the device 160.

Further, the means for differential counting of leukocytes 160 comprises one or more processors, working collectively or individually, optionally the counting means comprises the processor.

In one embodiment, the apparatus includes one or more processors working collectively or individually and one or more storage devices (not shown) storing programs for implementing respective steps in the method of differential white blood cell counting according to an embodiment of the present invention. The processor is used for operating the program stored in the storage device to execute the corresponding steps of the method for classifying and counting the white blood cells according to the embodiment of the invention.

In one embodiment of the invention, the program when executed by the processor causes the apparatus for differential white blood cell counting 160 to perform the steps of: obtaining an initial leukocyte histogram of leukocytes in a blood sample, wherein the initial leukocyte histogram is a histogram of the number and volume distribution of leukocytes; delineating a ghost region in the initial white blood cell histogram; correcting the ghost area to obtain a corrected lymphocyte area.

In one embodiment of the invention, the program when executed by the processor causes the apparatus for differential white blood cell counting 160 to perform the steps of: acquiring a first boundary line of a shadow area and a lymphocyte area in the initial leukocyte histogram, wherein the shadow area is an area with a volume smaller than the volume corresponding to the first boundary line so as to demarcate the shadow area in the initial leukocyte histogram.

In one embodiment of the invention, the program when executed by the processor causes the apparatus for differential white blood cell counting 160 to perform the steps of: demarcating a region of the initial leukocyte histogram having a volume less than a critical volume as the ghost region, the first boundary line being located at the critical volume.

In one embodiment of the invention, the program when executed by the processor causes the apparatus for differential white blood cell counting 160 to perform the steps of: obtaining a first peak point of the initial leukocyte histogram along the direction of volume increase starting from the minimum volume; determining the first boundary line based on the first peak point. More specifically, the following steps are performed: the first boundary line is disposed at a position spaced apart from the volume of the first peak point by a predetermined volume, and the volume corresponding to the first boundary line is larger than the volume of the first peak point.

In one embodiment of the invention, the program when executed by the processor causes the apparatus for differential white blood cell counting 160 to perform the steps of: and searching a critical point with the slope being greater than the threshold slope for the first time in the initial leukocyte histogram curve from the first peak point along the direction of volume increase, wherein the first boundary line is a straight line passing through the critical point and perpendicular to the horizontal axis of the coordinate system. Optionally, the threshold slope is less than or equal to zero.

In one embodiment of the invention, the program when executed by the processor causes the apparatus for differential white blood cell counting 160 to perform the steps of: obtaining a first valley point of the initial white blood cell histogram from a minimum volume along a direction of volume increase; determining the first boundary line based on the first valley point, wherein the first boundary line is disposed at a position spaced apart from a volume of the first valley point by a predetermined volume, and the volume corresponding to the first boundary line is smaller than the volume of the first valley point.

In one embodiment of the invention, the program when executed by the processor causes the apparatus for differential white blood cell counting 160 to perform the steps of: searching a first peak point on the initial leukocyte histogram from the maximum volume in the direction of volume reduction; the first boundary line is disposed at a position spaced apart from the first peak point by a predetermined volume, and a volume corresponding to the first boundary line is smaller than a volume of the first peak point.

In one embodiment of the invention, the program when executed by the processor causes the apparatus for differential white blood cell counting 160 to perform the steps of: based on the results of at least two white blood cell counts, a ghost region is delineated in the initial white blood cell histogram.

The two white blood cell counts include a first white blood cell count and a second white blood cell count, the histogram of the first white blood cell count is the initial white blood cell histogram, and in one embodiment of the invention, the program when executed by the processor causes the apparatus for differential white blood cell count 160 to perform the steps of: obtaining a threshold area of the ghost region based on an area of the histogram of the first white blood cell count and a threshold proportion coefficient of the ghost region in the histogram of the first white blood cell count, wherein the threshold proportion coefficient is obtained based on a result of the first white blood cell count and a result of the second white blood cell count; determining a first boundary line of a ghost region and a lymphocyte region in the initial leukocyte histogram based on the threshold area to delineate a ghost region in the initial leukocyte histogram, wherein the first boundary line is located where an area in the histogram of the first leukocyte count is greater than or equal to the threshold area.

Optionally, the amount of hemolytic agent used for the first white blood cell count is less than the amount of hemolytic agent used for the second white blood cell count.

SGhost/(SGhost+SWbc)＝(WBC1-WBC2)/WBC1

wherein SGhost + SWbc is an area of the histogram of the first white blood cell count, the result WBC1 of the first white blood cell count, the result WBC2 of the second white blood cell count, and the threshold scaling factor is a ratio of the difference between the result WBC1 of the first white blood cell count and the result WBC2 of the second white blood cell count to the result WBC1 of the first white blood cell count.

In one embodiment of the invention, the program when executed by the processor causes the apparatus for differential white blood cell counting 160 to perform the steps of: and removing the ghost area, and compensating the lymphocyte area to obtain a corrected lymphocyte area.

In one embodiment of the invention, the program when executed by the processor causes the apparatus for differential white blood cell counting 160 to perform the steps of: defining a compensation line for the lymphocyte region in a blank region of the initial leukocyte histogram excluding the shadow region to compensate for the lymphocyte region. In one example, the compensation line includes at least one of a straight line and a curved line. Optionally, the compensation line passes through a predetermined point located in the blank area and a critical point, the critical point being an intersection of a first boundary line between the lymphocyte region and the ghost region and a curve of the initial leukocyte histogram. Illustratively, the predetermined point is a point on the horizontal axis of a rectangular coordinate system in which the initial white blood cell histogram is located.

The compensation line is obtained based on a predetermined function equation, which when executed by the processor in one embodiment of the invention causes the means 160 for differential counting of white blood cells to perform the following steps: calculating a lymphocyte compensation particle number from each volume between the predetermined point and the critical point on the compensation line based on the function equation; compensating the lymphocyte region based on the summed number of lymphocyte compensation particles for each volume. Optionally, the function equation includes at least one of a first order function, a second order function, a polynomial function of the third order and more than the third order, an exponential function, and a logarithmic function. Illustratively, the quadratic function is a convex function or a concave function.

In one embodiment of the invention, the program when executed by the processor causes the apparatus for differential white blood cell counting 160 to perform the steps of: calculating a lymphocyte compensation particle number from each integral volume between the predetermined point and the critical point on the compensation line based on the function equation, wherein the lymphocyte compensation particle number is an integer.

In one embodiment of the invention, the program when executed by the processor causes the apparatus for differential white blood cell counting 160 to perform the steps of: pre-classifying the white blood cells based on the initial white blood cell histogram to obtain different types of white blood cell regions before correcting the ghost regions to obtain corrected lymphocyte regions, wherein the different types of white blood cell regions comprise lymphocyte regions, and the pre-classification is at least two classifications; after correcting the ghost area to obtain a corrected lymphocyte area, a corrected leukocyte histogram is obtained based on the corrected lymphocyte area.

In one embodiment of the invention, the program when executed by the processor causes the apparatus for differential white blood cell counting 160 to perform the steps of: obtaining a corrected leukocyte histogram based on the corrected lymphocyte region; classifying the white blood cells based on the corrected white blood cell histogram to obtain different types of white blood cell regions, wherein the different types of white blood cell regions comprise lymphocyte regions, and the classification is at least two classifications.

In one embodiment of the invention, the program when executed by the processor causes the apparatus for differential white blood cell counting 160 to perform the steps of: acquiring a first valley point in the modified white blood cell histogram curve from the minimum volume along the direction of volume increase; defining a second boundary line between the modified lymphocyte region and another leukocyte region having a larger volume than the lymphocyte based on the first valley point. Optionally, the first valley point is a valley point between a first peak point and a second peak point taken from the minimum volume in a direction of volume increase. Optionally, the second dividing line is spaced from the first valley point by a first predetermined volume, and the volume to which the second dividing line corresponds is less than the volume of the first valley point.

The other white blood cells include a first white blood cell type, a second white blood cell type and a third white blood cell type, wherein the second boundary line is a boundary line between the lymphocyte and the first white blood cell type, and when the program is executed by the processor, the apparatus 160 for differential counting of white blood cells performs the following steps: demarcating a third boundary line between the first type of white blood cell and the second type of white blood cell based on the first valley point and demarcating a fourth boundary line between the second type of white blood cell and the third type of white blood cell based on the first valley point or the second peak point. Optionally, the third demarcation line is spaced from the valley point by a second predetermined volume, the fourth demarcation line is spaced from the first valley point by a third predetermined volume, the third predetermined volume being greater than the second predetermined volume. Optionally, the volume corresponding to the third dividing line is greater than the volume of the first valley point, and the volume corresponding to the fourth dividing line is greater than the volume of the first valley point.

In one embodiment of the invention, the program when executed by the processor causes the apparatus for differential white blood cell counting 160 to perform the steps of: and carrying out filtering processing on the corrected leukocyte histogram.

In an embodiment of the present invention, the orthogonal coordinate system of the initial white blood cell histogram and the corrected white blood cell histogram has a cell volume as an abscissa and a cell particle number of different volumes as an ordinate, and when the program is executed by the processor, the device 160 for classifying and counting white blood cells performs the following steps: and calculating the total number of the cell particles included in the different types of leukocyte areas based on the sum of the number of the cell particles corresponding to each volume in the different types of leukocyte areas in the corrected leukocyte histogram.

In addition, the embodiment of the invention also provides a computer storage medium, and the computer storage medium is stored with the computer program. One or more computer program instructions may be stored on the computer-readable storage medium, which may be executed by a processor to implement the program instructions stored by the storage device to implement the functions of the embodiments of the invention (implemented by the processor) described herein and/or other desired functions, such as performing the corresponding steps of the method for white blood cell count classification according to the embodiments of the invention, and various applications and various data, such as various data used and/or generated by the applications, etc., may also be stored in the computer-readable storage medium.

For example, the computer storage medium may include, for example, a memory card of a smart phone, a storage component of a tablet computer, a hard disk of a personal computer, a Read Only Memory (ROM), an Erasable Programmable Read Only Memory (EPROM), a portable compact disc read only memory (CD-ROM), a USB memory, or any combination of the above storage media.

In addition, the embodiment of the present invention further provides a blood analyzer, which may include the above-described apparatus 160 for classifying white blood cell count according to the embodiment of the present invention, and a person skilled in the art can understand the structure, operation, and the like of the blood analyzer based on the above-described apparatus 160 for classifying white blood cell count, and therefore, in order to avoid repetition, the details are not repeated herein.

Blood analyzers are used for various analyses of blood components, such as counting and classifying leukocytes in blood, detecting the concentration of Hemoglobin (HGB) in erythrocytes, counting platelets, and the like. Therefore, the structure of the complete blood analyzer may include a device for detecting hemoglobin, a device for counting and detecting platelets, and the like, in addition to the aforementioned device for classifying white blood cell count.

In summary, in order to solve the problem of inaccurate counting and classifying of white blood cells caused by interference of cell debris on lymphocyte regions, particularly the problem of inaccurate counting and classifying of white blood cells caused by interference of cell debris on lymphocyte regions of a white blood cell histogram in an animal blood sample detection process, the method, the device and the blood analyzer for counting and classifying white blood cells according to the embodiment of the invention obtain an initial white blood cell histogram of the white blood cells in the blood sample, wherein the initial white blood cell histogram is a histogram of the number and volume distribution of the white blood cells; delineating a ghost region in the initial white blood cell histogram; and correcting the ghost area to obtain a corrected lymphocyte area, for example, reducing or even eliminating the interference of the ghost area to the lymphocyte area in a leukocyte histogram by compensating the lymphocyte area, and improving the accuracy of counting and classifying leukocytes, so that the actual condition of the blood sample is detected by more truly reflecting the counting and classifying result, and a doctor and the like can reasonably judge the health condition of the blood sample source sample according to the result to make reasonable medical diagnosis and the like.

Although the illustrative embodiments have been described herein with reference to the accompanying drawings, it is to be understood that the foregoing illustrative embodiments are merely exemplary and are not intended to limit the scope of the invention thereto. Various changes and modifications may be effected therein by one of ordinary skill in the pertinent art without departing from the scope or spirit of the present invention. All such changes and modifications are intended to be included within the scope of the present invention as set forth in the appended claims.

Those of ordinary skill in the art will appreciate that the various illustrative elements and algorithm steps described in connection with the embodiments disclosed herein may be implemented as electronic hardware or combinations of computer software and electronic hardware. Whether such functionality is implemented as hardware or software depends upon the particular application and design constraints imposed on the implementation. Skilled artisans may implement the described functionality in varying ways for each particular application, but such implementation decisions should not be interpreted as causing a departure from the scope of the present invention.

In the several embodiments provided in the present application, it should be understood that the disclosed apparatus and method may be implemented in other ways. For example, the above-described device embodiments are merely illustrative, and for example, the division of the units is only one logical functional division, and other divisions may be realized in practice, for example, a plurality of units or components may be combined or integrated into another device, or some features may be omitted, or not executed.

In the description provided herein, numerous specific details are set forth. It is understood, however, that embodiments of the invention may be practiced without these specific details. In some instances, well-known methods, structures and techniques have not been shown in detail in order not to obscure an understanding of this description.

Similarly, it should be appreciated that in the description of exemplary embodiments of the invention, various features of the invention are sometimes grouped together in a single embodiment, figure, or description thereof for the purpose of streamlining the invention and aiding in the understanding of one or more of the various inventive aspects. However, the method of the present invention should not be construed to reflect the intent: that the invention as claimed requires more features than are expressly recited in each claim. Rather, as the following claims reflect, inventive aspects lie in less than all features of a single disclosed embodiment. Thus, the claims following the detailed description are hereby expressly incorporated into this detailed description, with each claim standing on its own as a separate embodiment of this invention.

It will be understood by those skilled in the art that all of the features disclosed in this specification (including any accompanying claims, abstract and drawings), and all of the processes or elements of any method or apparatus so disclosed, may be combined in any combination, except combinations where such features are mutually exclusive. Each feature disclosed in this specification (including any accompanying claims, abstract and drawings) may be replaced by alternative features serving the same, equivalent or similar purpose, unless expressly stated otherwise.

Furthermore, those skilled in the art will appreciate that while some embodiments described herein include some features included in other embodiments, rather than other features, combinations of features of different embodiments are meant to be within the scope of the invention and form different embodiments. For example, in the claims, any of the claimed embodiments may be used in any combination.

The various component embodiments of the invention may be implemented in hardware, or in software modules running on one or more processors, or in a combination thereof. Those skilled in the art will appreciate that a microprocessor or Digital Signal Processor (DSP) may be used in practice to implement some or all of the functionality of some of the modules according to embodiments of the present invention. The present invention may also be embodied as apparatus programs (e.g., computer programs and computer program products) for performing a portion or all of the methods described herein. Such programs implementing the present invention may be stored on computer-readable media or may be in the form of one or more signals. Such a signal may be downloaded from an internet website or provided on a carrier signal or in any other form.

It should be noted that the above-mentioned embodiments illustrate rather than limit the invention, and that those skilled in the art will be able to design alternative embodiments without departing from the scope of the appended claims. In the claims, any reference signs placed between parentheses shall not be construed as limiting the claim. The invention may be implemented by means of hardware comprising several distinct elements, and by means of a suitably programmed computer. In the unit claims enumerating several means, several of these means may be embodied by one and the same item of hardware. The usage of the words first, second and third, etcetera do not indicate any ordering. These words may be interpreted as names.

Claims

1. A method of differential counting of leukocytes, the method comprising:

delineating a ghost region in the initial white blood cell histogram;

correcting the ghost area to obtain a corrected lymphocyte area.

2. The method of claim 1, wherein delineating a ghost region in the initial white blood cell histogram comprises:

3. The method of claim 2, wherein the method of delineating a ghost region in the initial white blood cell histogram comprises:

4. The method of claim 2, wherein obtaining a first boundary line of a ghost region and a lymphocyte region in the initial white blood cell histogram comprises:

determining the first boundary line based on the first peak point.

5. The method of claim 4, wherein said determining the first boundary line based on the first peak point comprises:

6. The method of claim 4, wherein said determining the first boundary line based on the first peak point comprises:

7. The method of claim 6, wherein the threshold slope is less than or equal to zero.

8. The method of claim 2, wherein obtaining a first boundary line of a ghost region and a lymphocyte region in the initial white blood cell histogram comprises:

9. The method of claim 2, wherein the method of delineating a ghost region in the initial white blood cell histogram comprises:

10. The method of claim 1, wherein the method of delineating a ghost region in the initial white blood cell histogram comprises:

11. The method of claim 10, wherein the two white blood cell counts include a first white blood cell count and a second white blood cell count, the histogram of the first white blood cell count being the initial white blood cell histogram, a ghost region being delineated in the initial white blood cell histogram based on results of the at least two white blood cell counts, comprising:

12. The method of claim 11, wherein the amount of hemolytic agent used for the first white blood cell count is less than the amount of hemolytic agent used for the second white blood cell count.

13. The method of claim 11, wherein the threshold area of the ghost region SGhost satisfies the following equation:

S_Ghost/(S_Ghost+S_Wbc)＝(WBC1-WBC2)/WBC1

14. The method of claim 1, wherein modifying the ghost region to obtain a modified lymphocyte region comprises:

15. The method of claim 14, wherein said compensating said lymphocyte region comprises:

16. The method of claim 15,

the compensation line passes through a predetermined point located in the blank region and a critical point, which is an intersection of a first boundary line between the lymphocyte region and the ghost region and a curve of the initial leukocyte histogram.

17. The method of claim 16, wherein the predetermined point is a point on a horizontal axis of a rectangular coordinate system in which the initial white blood cell histogram lies.

18. The method of claim 15, wherein the compensation line comprises at least one of a straight line and a curved line.

19. The method of claim 16, wherein the compensation line is obtained based on a predetermined function equation, the method further comprising:

20. The method of claim 19, wherein the function equation comprises at least one of a first order function, a second order function, a polynomial function of third and more than third, an exponential function, and a logarithmic function.

21. The method of claim 20, wherein the quadratic function is a convex function or a concave function.

22. The method of claim 19, wherein calculating a lymphocyte compensation particle number from each volume between the predetermined point and the critical point on the compensation line based on the function equation comprises:

23. The method of claim 1, wherein the method further comprises:

pre-classifying the white blood cells based on the initial white blood cell histogram to obtain different types of white blood cell regions before correcting the ghost regions to obtain corrected lymphocyte regions, wherein the different types of white blood cell regions comprise lymphocyte regions, and the pre-classification is at least two classifications;

after correcting the ghost area to obtain a corrected lymphocyte area, a corrected leukocyte histogram is obtained based on the corrected lymphocyte area.

24. The method of claim 1, wherein the method further comprises:

25. The method of claim 24, wherein said classifying the white blood cells based on the modified white blood cell histogram comprises:

26. The method of claim 25, wherein the first valley point is a valley point between a first peak point and a second peak point taken from a minimum volume in a direction of increasing volume.

27. The method of claim 25, wherein said second boundary line is spaced from said first valley point by a first predetermined volume and said second boundary line corresponds to a volume less than the volume of said first valley point.

28. The method of claim 26, wherein the other white blood cells include a first type of white blood cell, a second type of white blood cell, and a third type of white blood cell that sequentially increase in volume, wherein the second boundary line is a boundary line between the lymphocyte and the first type of white blood cell, and wherein classifying the white blood cells based on the modified white blood cell histogram further comprises:

29. The method of claim 28, wherein said third demarcation line is spaced from said first valley point by a second predetermined volume, and said fourth demarcation line is spaced from said first valley point by a third predetermined volume, said third predetermined volume being greater than said second predetermined volume.

30. The method of claim 28, wherein the third demarcation line corresponds to a volume greater than a volume of the first valley point and the fourth demarcation line corresponds to a volume greater than a volume of the first valley point.

31. The method of claim 24, further comprising, prior to the step of classifying the white blood cells based on the modified white blood cell histogram, the step of:

and carrying out filtering processing on the corrected leukocyte histogram.

32. The method of claim 23 or 24, wherein the orthogonal coordinate system in which the initial white blood cell histogram and the modified white blood cell histogram are located has a cell volume size as an abscissa and a cell particle number of different volumes as an ordinate, and wherein the method further comprises:

33. The method of claim 1, wherein the initial white blood cell histogram is obtained based on electrical impedance measurements of the blood sample.

34. The method of claim 1, wherein the white blood cells comprise animal white blood cells.

35. An apparatus for differential counting of leukocytes, the apparatus comprising one or more processors, working together or separately, for performing the method of any one of claims 1 to 34.

36. The apparatus of claim 35, wherein the apparatus further comprises:

37. A blood analyser comprising a device according to claim 35 or 36.