阅读说明：本技术 测定血小板浓度的方法及系统 (Method and system for determining platelet concentration ) 是由 祁欢 叶波 郑文波 胡长松 郁琦 李朝阳 于 2019-04-26 设计创作，主要内容包括：一种分析血液样本中血小板浓度的方法、血液分析系统(100)及存储介质,测定血液样本中血小板浓度的方法包括：将血液样本的第一份与稀释液混合,形成第一悬浮液；将血液样本的第二份与溶血剂和荧光染料混合以溶解红细胞和染色白细胞,形成第二悬浮液；测量第一悬浮液流过小孔的直流阻抗信号；测量第二悬浮液流过光学流动室(20)的光散射信号和荧光信号；分析第一悬浮液的该直流阻抗信号以获取第一血小板分布；分析第二悬浮液的光散射信号和荧光信号以区分血小板和白细胞并获取第二血小板分布；基于第一血小板分布和第二血小板分布确定血液样本的血小板分析数据,如血小板浓度。(A method of analyzing a blood sample for platelet concentration, a blood analysis system (100) and a storage medium, the method of determining the concentration of platelets in a blood sample comprising: mixing a first portion of the blood sample with a diluent to form a first suspension; mixing a second portion of the blood sample with a hemolytic agent and a fluorescent dye to lyse red blood cells and stain white blood cells to form a second suspension; measuring a DC impedance signal of the first suspension flowing through the orifice; measuring light scatter and fluorescence signals of the second suspension flowing through the optical flow cell (20); analyzing the dc impedance signal of the first suspension to obtain a first platelet distribution; analyzing the light scattering signal and the fluorescence signal of the second suspension to distinguish platelets from leukocytes and obtain a second platelet distribution; platelet analysis data, such as platelet concentration, of the blood sample is determined based on the first platelet distribution and the second platelet distribution.)

A method of determining the concentration of platelets in a blood sample, comprising:

mixing a first portion of the blood sample with a diluent to form a first suspension;

mixing a second portion of the blood sample with a hemolytic agent and a fluorescent dye to lyse red blood cells and stain white blood cells to form a second suspension;

measuring a direct current impedance signal of the first suspension flowing through the orifice;

measuring light scatter and fluorescence signals of the second suspension flowing through the optical flow cell;

analyzing the dc impedance signal of the first suspension to obtain a first platelet distribution;

analyzing the light scatter signal and the fluorescence signal of the second suspension to distinguish platelets from leukocytes and/or nucleated red blood cells and obtain a second platelet distribution; and

determining a platelet concentration of the blood sample based on the first platelet distribution and the second platelet distribution.

The method of claim 1, wherein said distinguishing platelets from white blood cells and/or nucleated red blood cells comprises distinguishing a region of platelets from a region of white blood cells and/or nucleated red blood cells in a light scatter and fluorescence scatter plot obtained from said second suspension.

The method of claim 2, wherein the first platelet distribution is a platelet dc impedance histogram obtained from the first suspension.

The method of claim 3, wherein the second platelet distribution is a derived platelet volume histogram generated using light scattering signals of platelets in the platelet region.

The method of claim 4, wherein the step of determining the platelet concentration of the blood sample comprises generating a fused platelet histogram using the platelet DC impedance histogram from the first suspension and the derived platelet volume histogram from the second suspension, and obtaining the platelet concentration based on the fused platelet histogram.

The method of claim 5, further comprising displaying to a user the platelet concentration, the platelet DC impedance histogram from the first suspension, the derived platelet volume histogram from the second suspension, the fused platelet histogram, a superposition of the platelet DC impedance histogram from the first suspension and the derived platelet volume histogram from the second suspension, a superposition of the platelet DC impedance histogram from the first suspension and the fused platelet histogram, the platelet region of the light scatter and fluorescence scatter plot from the second suspension, or a combination thereof.

The method of claim 3, wherein the second platelet distribution is a two-dimensional distribution of platelets in a platelet region in a forward light scatter and fluorescence scatter plot obtained from the second suspension.

The method of claim 7, wherein the step of determining the platelet concentration of the blood sample comprises:

determining a platelet trough to peak ratio of the platelet dc impedance histogram of the first suspension;

determining a number of events for a specified one of the platelet regions of the forward light scatter and fluorescence scatter plot from the second suspension;

determining a derivative separation threshold for valleys between platelets and red blood cells in the platelet dc impedance histogram based on the platelet valley to peak ratio and the number of events for the specified region; and

and distinguishing the platelets and red blood cells in the platelet direct current impedance histogram by using the derivative separation threshold value to obtain the platelet concentration of the blood sample.

The method of claim 8, further comprising displaying the platelet concentration, the platelet region in the forward light scatter and fluorescence scatter plot from the second suspension, the designated region in the platelet region, a number of events of the designated region, the platelet trough to peak ratio in the platelet dc impedance histogram from the first suspension, the platelet dc impedance histogram from the first suspension with the derivative separation threshold, or a combination thereof to a user.

The method of claim 1, wherein the step of determining the platelet concentration of the blood sample comprises: generating a third platelet distribution based on the first platelet distribution and the second platelet distribution, and obtaining a platelet concentration based on the third platelet distribution.

The method of claim 1, further comprising differentiating leukocytes in the blood sample into subpopulations of leukocytes based on the light scatter signal and the fluorescence signal of the second suspension, preferably wherein the differentiating leukocytes in the blood sample into subpopulations of leukocytes comprises:

differentiating monocytes, lymphocytes, neutrophils and eosinophils; or

Basophils were differentiated.

The method of claim 1,

further comprising performing a white blood cell count of the blood sample on the second suspension; or

Identifying nucleated red blood cells or immature white blood cells in the second suspension.

A blood analysis system for determining the concentration of platelets in a blood sample, comprising:

a first module comprising a first mixing chamber for mixing a first portion of the blood sample with a diluent to form a first suspension, and a dc impedance detector mounted to an orifice of a flow path communicating with the first mixing chamber, the dc impedance detector for detecting a dc impedance signal of the first suspension through the orifice;

a second module comprising a second mixing chamber for mixing a second portion of the blood sample with a hemolysing agent and a fluorescent dye, lysing red blood cells, and staining white blood cells to form a second suspension, a light source for directing a light beam at a detection well of an optical flow chamber in communication with the second mixing chamber, and at least one optical detector mounted to the optical flow chamber for detecting light scatter and fluorescence signals of the second suspension passing through the detection well of the optical flow chamber; and

a data processing module operatively connected to the DC impedance detector of the first module and the at least one optical detector of the second module, respectively, the data processing module comprising a processor and a non-transitory computer-readable storage medium programmed with a computer application that, when executed by the processor, causes the processor to generate a first platelet distribution based on the DC impedance signal of the first suspension, distinguish platelets from leukocytes and/or nucleated red blood cells based on the light scatter signal and the fluorescence signal of the second suspension, generate a second platelet distribution, determine a platelet concentration of the blood sample based on the first platelet distribution and the second platelet distribution.

The blood analysis system of claim 13, wherein the first platelet distribution is a platelet dc impedance histogram derived from the first suspension, and the second platelet distribution is a derived platelet volume histogram generated based on light scattering from the second suspension and light scattering signals from platelets in a platelet region in a fluorescence scattergram.

The blood analysis system of claim 14, wherein the computer application of the data processing module, when executed by the processor, causes the processor to generate a fused platelet histogram using a platelet dc impedance histogram from the first suspension and the derived platelet volume histogram from the second suspension, and obtain a platelet concentration based on the fused platelet histogram.

The blood analysis system of claim 15, further comprising a user interface operatively coupled to the data processing module and configured to display the platelet concentration, the platelet dc impedance histogram from the first suspension, the derived platelet volume histogram from the second suspension, the fused platelet histogram, a superimposed plot of the platelet dc impedance histogram from the first suspension and the derived platelet volume histogram from the second suspension, a superimposed plot of the platelet dc impedance histogram from the first suspension and the fused platelet histogram, the platelet region of the light scatter and fluorescence scatter plot from the second suspension, or a combination thereof.

The blood analysis system of claim 13, wherein the first platelet distribution is a platelet dc impedance histogram from the first suspension and the second platelet distribution is a two-dimensional distribution of platelets in a platelet region in a forward light scatter and fluorescence scatter plot from the second suspension.

The blood analysis system of claim 17, wherein the computer application of the data processing module, when executed by the processor, causes the processor to

Determining a platelet trough to peak ratio of the platelet dc impedance histogram of the first suspension;

determining a number of events for a specified one of the platelet regions of the forward light scatter and fluorescence scatter plot from the second suspension;

determining a derivative separation threshold for valleys between platelets and red blood cells in the platelet dc impedance histogram based on the platelet valley to peak ratio and the number of events for the specified region; and

and distinguishing the platelets and red blood cells in the platelet direct current impedance histogram by using the derivative separation threshold value to obtain the platelet concentration of the blood sample.

The blood analysis system of claim 13, wherein the step of determining the platelet concentration of the blood sample comprises: generating a third platelet distribution based on the first platelet distribution and the second platelet distribution, and obtaining a platelet concentration based on the third platelet distribution.

The blood analysis system of claim 13, wherein the computer application of the data processing module, when executed by the processor, causes the processor to distinguish leukocytes in the blood sample into subpopulations of leukocytes based on the light scatter signal and the fluorescence signal of the second suspension, preferably the distinguishing of leukocytes in the blood sample into subpopulations of leukocytes comprises:

differentiating monocytes, lymphocytes, neutrophils and eosinophils; or

Basophils were differentiated.

The blood analysis system of claim 13,

further comprising performing a white blood cell count of the blood sample on the second suspension; or

Identifying nucleated red blood cells or immature white blood cells in the second suspension.

A method of determining the concentration of platelets in a blood sample, comprising:

mixing a first portion of the blood sample with a diluent to form a first suspension;

mixing a second portion of the blood sample with a hemolysing agent to lyse red blood cells to form a second suspension;

measuring a direct current impedance signal of the first suspension flowing through the orifice;

measuring a forward light scatter signal and a side light scatter signal or a mid-angle light scatter signal of the second suspension flowing through the optical flow cell;

analyzing the dc impedance signal of the first suspension to obtain a first platelet distribution;

analyzing the forward light scatter signal and the side light scatter signal or the mid-angle light scatter signal of the second suspension to distinguish between platelets and leukocytes and obtain a second platelet distribution; and

determining a platelet concentration of the blood sample based on the first platelet distribution and the second platelet distribution.

The method of claim 22, wherein distinguishing between platelets and leukocytes comprises distinguishing between platelet regions and leukocyte regions in a forward light scatter signal and side light scatter signal scatter plot or a forward light scatter signal and medium angle light scatter signal scatter plot obtained from the second suspension.

The method of claim 23, wherein the first platelet distribution is a platelet dc impedance histogram obtained from the first suspension.

The method of claim 24, wherein the second platelet distribution is a derived platelet volume histogram generated using light scattering signals of platelets in the platelet region.

The method of claim 25, wherein the step of determining the platelet concentration of the blood sample comprises generating a fused platelet histogram using the platelet dc impedance histogram from the first suspension and the derived platelet volume histogram from the second suspension, and obtaining the platelet concentration based on the fused platelet histogram.

The method of claim 24, wherein the second platelet distribution is a two-dimensional distribution of platelets in a platelet region in the forward and side or mid-angle light scatter signal scatter plot from the second suspension.

The method of claim 27, wherein the step of determining the platelet concentration of the blood sample comprises:

determining a platelet trough to peak ratio of the platelet dc impedance histogram of the first suspension;

determining a number of events for a specified one of the platelet regions from the forward and side or mid-angle light scatter signal scatter plots of the forward and side light scatter signals obtained from the second suspension;

determining a derivative separation threshold for valleys between platelets and red blood cells in the platelet dc impedance histogram based on the platelet valley to peak ratio and the number of events for the specified region; and

and distinguishing the platelets and red blood cells in the platelet direct current impedance histogram by using the derivative separation threshold value to obtain the platelet concentration of the blood sample.

The method of claim 22, wherein the step of determining the platelet concentration of the blood sample comprises: generating a third platelet distribution based on the first platelet distribution and the second platelet distribution, and obtaining a platelet concentration based on the third platelet distribution.

The method of claim 22, further comprising differentiating leukocytes in the blood sample into subpopulations of leukocytes based on the light scatter signal of the second suspension.

The method of claim 30, wherein the differentiating the white blood cells in the blood sample into subpopulations of white blood cells comprises:

differentiating monocytes, lymphocytes, neutrophils and eosinophils; or

Distinguishing basophils; or

Performing a white blood cell count of the blood sample on the second suspension.

A blood analysis system for determining the concentration of platelets in a blood sample, comprising:

a first module comprising a first mixing chamber for mixing a first portion of the blood sample with a diluent to form a first suspension, and a dc impedance detector mounted to an orifice of a flow path communicating with the first mixing chamber, the dc impedance detector for detecting a dc impedance signal of the first suspension through the orifice;

a second module comprising a second mixing chamber for mixing a second portion of the blood sample with a hemolysing agent, lysing red blood cells to form a second suspension, a light source for directing a light beam at a detection aperture of an optical flow chamber in communication with the second mixing chamber, and at least one optical detector mounted to the optical flow chamber for detecting a forward light scatter signal and a side light scatter signal or a mid-angle light scatter signal of the second suspension passing through the detection aperture of the optical flow chamber; and

a data processing module operatively connected to the DC impedance detector of the first module and the at least one optical detector of the second module, respectively, the data processing module comprising a processor and a non-transitory computer-readable storage medium programmed with a computer application that, when executed by the processor, causes the processor to generate a first platelet distribution based on the DC impedance signal of the first suspension, distinguish platelets from leukocytes based on the light scatter signal and the side or mid-angle light scatter signal of the second suspension, generate a second platelet distribution, determine a platelet concentration of the blood sample based on the first platelet distribution and the second platelet distribution.

The blood analysis system of claim 32, wherein the first platelet distribution is a platelet dc impedance histogram obtained from the first suspension.

The blood analysis system of claim 33, wherein the second platelet distribution is a derived platelet volume histogram generated based on the forward and side light scatter signal scatter plots or the light scatter signals of platelets in a platelet region in the forward and mid-angle light scatter signal scatter plot obtained for the second suspension.

The blood analysis system of claim 34, wherein the computer application of the data processing module, when executed by the processor, causes the processor to generate a fused platelet histogram using a platelet dc impedance histogram from the first suspension and the derived platelet volume histogram from the second suspension, and obtain a platelet concentration based on the fused platelet histogram.

The blood analysis system of claim 33, wherein the second platelet distribution is a two-dimensional distribution of platelets in a platelet region in a forward and side or mid-angle light scatter signal scatter plot of forward and side light scatter signals obtained from the second suspension.

The blood analysis system of claim 36, wherein the computer application of the data processing module, when executed by the processor, causes the processor to

Determining a platelet trough to peak ratio of the platelet dc impedance histogram of the first suspension;

determining a number of events for a specified one of the platelet regions from the forward and side or mid-angle light scatter signal scatter plots of the forward and side light scatter signals obtained from the second suspension;

determining a derivative separation threshold for valleys between platelets and red blood cells in the platelet dc impedance histogram based on the platelet valley to peak ratio and the number of events for the specified region; and

and distinguishing the platelets and red blood cells in the platelet direct current impedance histogram by using the derivative separation threshold value to obtain the platelet concentration of the blood sample.

The blood analysis system of claim 32, wherein the step of determining the platelet concentration of the blood sample comprises: generating a third platelet distribution based on the first platelet distribution and the second platelet distribution, and obtaining a platelet concentration based on the third platelet distribution.

The blood analysis system of claim 32, wherein the computer application of the data processing module, when executed by the processor, causes the processor to distinguish leukocytes in the blood sample into subpopulations of leukocytes based on the light scatter signal of the second suspension.

The blood analysis system of claim 32, wherein the computer application of the data processing module, when executed by the processor, causes the processor to distinguish leukocytes from monocytes, lymphocytes, neutrophils, and eosinophils or from basophils in the blood sample based on the light scatter signal of the second suspension; or

Performing a white blood cell count of the blood sample on the second suspension.

A method of analyzing a blood sample, comprising:

obtaining a direct current impedance signal of a first suspension of the blood sample;

obtaining at least two optical signals of the second suspension of the blood sample, the at least two optical signals including a forward light scatter signal and a first optical signal for providing information on cell content;

obtaining a first platelet distribution based on the DC impedance signal;

obtaining a second platelet distribution based on the at least two optical signals; and

obtaining a third platelet distribution based on the first platelet distribution and the second platelet distribution;

the first suspension is formed by mixing a first portion of the blood sample with a diluent, and the second suspension is formed by mixing a second portion of the blood sample with a treatment reagent that includes a hemolysing agent to lyse red blood cells in the second portion of the blood sample.

The method of analyzing a blood sample of claim 41, further comprising: outputting at least one of the first, second, and third platelet distributions.

The method of analyzing a blood sample of claim 41, wherein the step of obtaining a second platelet distribution comprises:

obtaining a scatter diagram of the second suspension based on the at least two optical signals; and

in the scattergram, platelets are distinguished from leukocytes and/or nucleated erythrocytes, resulting in a platelet region.

The method of analyzing a blood sample of claim 43, wherein the step of obtaining a second platelet distribution further comprises: obtaining a derived platelet volume histogram of the blood sample based on the scattered optical signals of the platelet region.

The method of analyzing a blood sample of claim 44, wherein the first platelet profile comprises a platelet DC impedance histogram of the blood sample; the step of obtaining a third platelet distribution comprises: and acquiring a fused platelet histogram according to a preset condition based on the platelet direct current impedance histogram and the derived platelet volume histogram.

The method of analyzing a blood sample of claim 45, wherein the step of obtaining a third platelet distribution further comprises: and acquiring platelet analysis data of the blood sample based on the fused platelet histogram, wherein the platelet analysis data is selected from one or more of platelet count, average platelet volume and platelet volume distribution width.

The method of analyzing a blood sample of claim 46, further comprising: displaying at least one of the platelet region, the derived platelet volume histogram, the overlay of the platelet DC impedance histogram and the derived platelet volume histogram, the fused platelet histogram, the overlay of the platelet DC impedance histogram and the fused platelet histogram, and the platelet analysis data.

The method of analyzing a blood sample of claim 43, wherein the step of obtaining a second platelet distribution further comprises: and acquiring the event number of a specified area in the platelet area.

The method of analyzing a blood sample of claim 48, wherein the first platelet profile comprises a platelet DC impedance histogram of the blood sample; the step of obtaining a third platelet distribution comprises:

determining a platelet trough to peak ratio of the platelet DC impedance histogram; and

and determining a derivative separation threshold value based on the platelet valley peak ratio and the number of events in the specified area, distinguishing platelets from red blood cells by using the derivative separation threshold value in the platelet direct current impedance histogram, and acquiring platelet analysis data of the blood sample, wherein the platelet analysis data is selected from one or more of platelet count, average platelet volume and platelet distribution width.

The method of analyzing a blood sample of claim 49, further comprising: displaying at least one of the platelet region, the designated region, the platelet trough to peak ratio, the platelet DC impedance histogram including the derivative separation threshold, the platelet analysis data.

The method of analyzing a blood sample of any of claims 41-50, wherein the first optical signal is selected from at least one of a side light scatter signal and a mid-angle light scatter signal.

The method of analyzing a blood sample of any one of claims 41-50, wherein the processing reagent further comprises a fluorescent dye for staining white blood cells, and the first optical signal is selected from at least one of a fluorescent signal, a side light scatter signal, and a mid-angle light scatter signal.

The method of analyzing a blood sample of claim 51 or 52, further comprising: differentiating leukocytes in the blood sample into subpopulations of leukocytes based on the at least two optical signals of the second suspension.

The method of analyzing a blood sample of claim 51 or 52, wherein leukocytes in the blood sample are differentiated from monocytes, lymphocytes, neutrophils and eosinophils or from basophils based on the at least two optical signals of the second suspension; or

Performing a white blood cell count of the blood sample on the second suspension; or

Identifying nucleated red blood cells or immature white blood cells in the second suspension.

A non-transitory computer-readable storage medium having a computer application program stored thereon, the computer application program comprising: the computer application, when executed by a processor, performs the steps of the method of analyzing a blood sample according to any one of claims 41-54.

A blood analysis system comprising:

a first module comprising a first mixing chamber for mixing a first portion of the blood sample with a diluent to form a first suspension, and a dc impedance detector mounted to an orifice of a flow path communicating with the first mixing chamber, the dc impedance detector for detecting a dc impedance signal of the first suspension through the orifice;

a second module comprising a second mixing chamber for mixing a second portion of the blood sample with a processing reagent to lyse red blood cells in the second portion of the blood sample to form a second suspension, a light source for directing a light beam at a detection well of an optical flow chamber in communication with the second mixing chamber, and at least one optical detector mounted to the optical flow chamber for detecting at least two optical signals of the second suspension passing through the detection well of the optical flow chamber, the at least two optical signals including a forward light scatter signal and a first optical signal for providing cell content information; and

a data processing module operatively connected to the DC impedance detector of the first module and the at least one optical detector of the second module, respectively, the data processing module including a processor and a non-transitory computer readable storage medium programmed with a computer application that, when executed by the processor, acquires a first platelet distribution based on the DC impedance signals, a second platelet distribution based on the at least two optical signals, and a third platelet distribution of the blood sample based on the first platelet distribution and the second platelet distribution.

The blood analysis system of claim 56, further comprising a user interface operatively coupled to the data processing module and configured to output at least one of the first platelet distribution, the second platelet distribution, and the third platelet distribution.

The blood analysis system of claim 56, wherein the data processing module obtaining a second platelet distribution comprises:

obtaining a scatter diagram of the second suspension based on the at least two optical signals; and

platelets are distinguished from leukocytes in the scatter plot, resulting in platelet regions.

The blood analysis system of claim 58, wherein the step of the data processing module obtaining a second platelet distribution further comprises: obtaining a derived platelet volume histogram of the blood sample based on the at least two optical signals of the platelet region.

The blood analysis system of claim 59, wherein the first platelet distribution comprises a platelet DC impedance histogram of the blood sample; the step of the data processing module acquiring a third platelet distribution comprises: and acquiring a fused platelet histogram according to a preset condition based on the platelet direct current impedance histogram and the derived platelet volume histogram.

The blood analysis system of claim 60, wherein the data processing module obtaining a third platelet distribution further comprises: and acquiring platelet analysis data of the blood sample based on the fused platelet histogram, wherein the platelet analysis data is selected from one or more of platelet count, average platelet volume and platelet volume distribution width.

The blood analysis system of claim 61, further comprising a user interface operatively coupled to the data processing module and configured to display at least one of the platelet region, the derived platelet volume histogram, the overlay of the platelet DC impedance histogram and the derived platelet volume histogram, the fused platelet histogram, the overlay of the platelet DC impedance histogram and the fused platelet histogram, and the platelet analysis data.

The blood analysis system of claim 58, wherein the step of obtaining a second platelet distribution further comprises: and acquiring the event number of a specified area in the platelet area.

The blood analysis system of claim 63, wherein the first platelet distribution comprises a platelet DC impedance histogram of the blood sample; the step of obtaining a third platelet distribution comprises:

determining a platelet trough to peak ratio of the platelet DC impedance histogram; and

and determining a derivative separation threshold value based on the platelet valley peak ratio and the number of events in the specified area, distinguishing platelets from red blood cells by using the derivative separation threshold value in the platelet direct current impedance histogram, and acquiring platelet analysis data of the blood sample, wherein the platelet analysis data is selected from one or more of platelet count, average platelet volume and platelet volume distribution width.

The blood analysis system of claim 64, further comprising a user interface operatively connected to the data processing module and configured to display at least one of the platelet region, the specified region, the platelet trough to peak ratio, the platelet DC impedance histogram including the derivative separation threshold, and the platelet analysis data.

The blood analysis system of any one of claims 56-65, wherein the first optical signal is selected from at least one of a side light scatter signal and a mid-angle light scatter signal.

The blood analysis system of any one of claims 56-65, wherein the processing reagent further comprises a fluorescent dye for staining leukocytes, and the first optical signal is selected from at least one of a fluorescent signal, a side light scatter signal, and a mid-angle light scatter signal.

The blood analysis system of claim 67, further comprising: differentiating the leukocytes in the blood sample into subpopulations of leukocytes based on the at least two optical signals of the second suspension, preferably the step of differentiating the leukocytes in the blood sample into subpopulations of leukocytes comprises:

differentiating monocytes, lymphocytes, neutrophils and eosinophils; or to differentiate basophils.

The blood analysis system of claim 68,

further comprising performing a white blood cell count of the blood sample on the second suspension; or recognizing nucleated red blood cells or immature leukocytes in the second suspension.