CN113196039A

CN113196039A - Sample optical detection device, sample detection method and sample analyzer

Info

Publication number: CN113196039A
Application number: CN201980080985.4A
Authority: CN
Inventors: 汪东生
Original assignee: Shenzhen Mindray Bio Medical Electronics Co Ltd
Current assignee: Shenzhen Mindray Bio Medical Electronics Co Ltd
Priority date: 2019-01-14
Filing date: 2019-06-11
Publication date: 2021-07-30
Also published as: WO2020146967A1; WO2020147255A1

Abstract

An optical sample detection device comprising: the flow chamber is used for allowing cells in a sample to be detected to pass through one by one; a light source for illuminating cells passing through the flow cell; a first forward scattered light signal collection assembly for collecting forward scattered light of a first angular range, the forward scattered light being light generated by the light source illuminating cells passing through the flow cell and reflected; a second forward scattered light signal collection assembly for directly collecting forward scattered light of a second angular range, the forward scattered light being light generated by the light source illuminating cells passing through the flow cell. The invention also relates to a sample detection method and a sample analyzer.

Description

Sample optical detection device, sample detection method and sample analyzer

Technical Field

The invention relates to a sample optical detection device, a sample detection method and a sample analyzer.

Background

Most of blood cell analyzers adopt a laser scattering principle to measure, and the principle is as follows: the cells are classified and counted by irradiating the cells with laser light and collecting forward scattered light, side scattered light (90-degree scattered light), and side fluorescent light (90-degree fluorescent light) generated after the cells are irradiated.

FIG. 1 is a diagram of an optical detection device of a blood cell analyzer, in which cells are passed through a flow chamber one by one under the action of a sheath flow, and when light emitted from a laser light source is collimated by a lens and then irradiated to the cells passing through the flow chamber, the light irradiated to the cells is scattered all around, and after the forward scattered light is collected by a collecting lens, an aperture is used to define the angle of the forward scattered light finally reaching a photodetector, for example, the forward scattered light is defined as a low-angle (or small-angle) forward scattered light, which is generally used for measuring the volume of the cells; meanwhile, lateral light is collected through another collecting lens in the direction perpendicular to light rays irradiating cells, the collected lateral light is reflected and refracted through a dichroic mirror, lateral scattered light in the lateral light is reflected when passing through the dichroic mirror and then reaches a corresponding photoelectric detector, the lateral scattered light is generally used for measuring the surface complexity of the cells, lateral fluorescence in the lateral light is refracted or transmitted and then also reaches the corresponding photoelectric detector through an optical filter, and the lateral fluorescence is generally used for measuring the intracellular nucleic acid content.

Technical problem

The optical detection device in fig. 1 has only three measurement channels, namely, a low-angle forward scattering light channel, a side scattering light channel and a side fluorescence channel, and therefore, cells can only be classified and counted based on signals acquired by the three measurement channels, which limits further cell scoring and counting to some extent, i.e., cannot perform more-dimensional and more-detailed classification and counting, and reduces the classification capability of abnormal cells; if a technician replaces or adds a high-angle (or large-angle) scattered light channel to the low-angle forward scattered light channel in fig. 1, the technician can directly use the target surface of the photodetector to receive the large-angle forward scattered light, but the signal to noise ratio of the received signal is very poor, so in order to ensure the signal quality, the technician usually uses a plurality of complex lens assemblies to collect the large-angle forward scattered light and then emits the large-angle forward scattered light to the corresponding photodetector, which greatly increases the cost of the device; in addition, the optical detection device is generally large in size because the optical path structure is selected, for example, the forward scattering light channel is generally designed as a refractive optical path structure, which results in large size of the optical detection device, especially when the forward scattering light channel is used for collecting scattered light in multiple angular ranges (e.g., low angle, high angle, etc.).

In addition, at present, when RBC and PLT are measured in blood cell analysis, an impedance method or an optical method is generally used. However, both of the above two methods have some disadvantages, for example, for some abnormal samples, such as low value PLT samples, the boundary between the PLT histogram and the RBC histogram often has no obvious boundary in the impedance method, so that the algorithm cannot accurately cut the PLT and RBC histograms, and further cannot obtain an accurate PLT measurement result. The accuracy of low-value PLT is a blood routine index which is of great clinical concern, and the low-value PLT becomes a main defect of impedance measurement. In addition, in the optical method, a fluorescent dye is required to stain cells, and special diluent is required to perform sphericization treatment on the cells, so that the use cost is high, and the optical method is not favorable for clinical popularization.

Technical solution

The present invention mainly provides a sample optical detection device, which is described below.

An embodiment of a sample optical detection device includes:

the flow chamber is used for allowing cells in a sample to be detected to pass through one by one;

a light source for illuminating cells passing through the flow cell;

a first forward scattered light signal collection assembly for collecting forward scattered light of a first angular range, the forward scattered light being light generated by the light source illuminating cells passing through the flow cell and reflected;

a second forward scattered light signal collection assembly for directly collecting forward scattered light of a second angular range, the forward scattered light being light generated by the light source illuminating cells passing through the flow cell.

In one embodiment, the optical sample detection device further comprises a reflective light blocking assembly disposed on an optical path of forward scattered light generated by the light source irradiating the cells passing through the flow cell, for reflecting the forward scattered light of the first angular range to the first forward scattered light signal collection assembly, and allowing the forward scattered light of the second angular range to be directly incident to the second forward scattered light signal collection assembly.

In one embodiment, the reflective light barrier assembly includes a mirror for reflecting forward scattered light of a first angular range to the first forward scattered light signal collection assembly.

In one embodiment, the reflector is elliptical.

In one embodiment, the reflective light blocking assembly further comprises a light shielding bar for shielding stray light except for the forward scattered light in the second angle range; the reflector is arranged on the shading strip.

In one embodiment, the reflector is disposed on the light-shielding strip in a manner that a long axis of the reflector coincides with the light-shielding strip.

In one embodiment, the optical sample detection device further includes a lens assembly disposed on an optical path of forward scattered light generated by the cell passing through the flow cell irradiated by the light source, and configured to collect the forward scattered light in the first angle range and the second angle range and emit the collected forward scattered light to the reflective light blocking assembly.

In one embodiment, the lens assembly includes one aspheric lens and one spherical lens, or a plurality of aspheric lenses, or a plurality of spherical lenses, or one aspheric lens and a plurality of spherical lenses.

In one embodiment, when the lens assembly comprises a spherical lens, the effective data aperture of the spherical lens closest to the flow cell is at least 0.34.

In an embodiment, the sample optical detection apparatus further includes a straight blocking diaphragm disposed between the lens assembly and the reflective light blocking assembly, for blocking direct angle light in the forward scattered light emitted from the lens assembly, and/or limiting the forward scattered light emitted from the lens assembly to the first angle range and the second angle range.

In one embodiment, the first forward scattered light signal collection assembly comprises a first angle limiting diaphragm, a stray light blocking diaphragm and a photodetector which are arranged in sequence; the first angle limiting diaphragm is used for limiting the reflected forward scattered light in a first angle range and converging the reflected forward scattered light in the stray light blocking diaphragm, the stray light blocking diaphragm is used for shielding stray light of the forward scattered light in the first angle range, and the photoelectric detector is used for converting the collected forward scattered light in the first angle range into an electric signal.

In an embodiment, the second forward scattered light signal collecting assembly includes a stray light blocking diaphragm and a photodetector, which are sequentially disposed, the stray light blocking diaphragm is configured to block stray light of the forward scattered light of the second angle range, and the photodetector is configured to convert the collected forward scattered light of the second angle range into an electrical signal.

In one embodiment, the optical sample detection device further comprises a third forward scattered light signal collection assembly for collecting forward scattered light of a third angular range, the forward scattered light being light generated by the light source illuminating the cells passing through the flow cell and having undergone at least one reflection.

In one embodiment, the third forward scattered light signal collection assembly includes a reflector, a third angular range aperture stop, and a photodetector, which are sequentially disposed; the reflector is used for reflecting the forward scattering light of a third angle range generated by cells irradiated by the light source through the flow chamber to the third angle range aperture stop, the third angle range aperture stop is used for limiting the forward scattering light in the third angle range, and the photoelectric detector is used for converting the collected forward scattering light in the third angle range into an electric signal.

In an embodiment, the first angle range is a low angle range, and/or the second angle range is a medium angle range, and/or the third angle range is a high angle range.

In one embodiment, the first angle range and the second angle range are continuous ranges.

In one embodiment, the first angle range, the second angle range and the third angle range are continuous ranges.

In one embodiment, the first angle range is 0 to 10 degrees or 1 to 10 degrees; and/or, the second angle ranges from 10 degrees to 20 degrees; and/or the third angle range is 20 degrees to 70 degrees.

In one embodiment, the optical sample detection device further comprises a light source shaping component for collimating and focusing the light beam emitted by the light source on the cell passing through the flow chamber.

In one embodiment, the light source shaping assembly includes a collimating lens and a first cylindrical mirror, which are sequentially disposed, the collimating lens is configured to collimate the light beam emitted by the light source, and the first cylindrical mirror is configured to converge the light beam to the center of the flow chamber in a direction in which the cell passes through.

In one embodiment, the sample optical detection apparatus further includes a second cylindrical mirror disposed on an exit path of the first cylindrical mirror, and configured to converge the light beam in a direction perpendicular to a cell passing direction, so that scattered light is irradiated into the straight stop.

In an embodiment, the optical detection device further includes an optical isolator disposed between the collimating lens and the first cylindrical mirror, for suppressing the feedback light.

In one embodiment, the optical sample detection device further comprises:

a side scatter signal collection assembly for collecting side scatter light generated by the light source illuminating cells passing through the flow cell; and/or the presence of a gas in the gas,

a lateral fluorescence signal collection assembly for collecting lateral fluorescence generated by the light source illuminating cells passing through the flow cell.

According to the sample optical detection device of the above embodiment, the first forward scattered light signal collecting element collects the reflected forward scattered light in the first angular range, and the second forward scattered light signal collecting element directly collects or collects the refracted or transmitted forward scattered light.

In addition, the embodiment of the invention also provides a sample detection method for detecting red blood cells and platelets in a blood sample, which comprises the following steps:

preparing a sample solution to be detected containing a blood sample to be detected and a diluent;

irradiating the sample liquid to be detected with light in an optical detection area;

collecting at least two scattered light signals generated by the particles in the sample liquid to be detected due to light irradiation;

and classifying the red blood cells and the platelets in the blood sample to be detected according to the at least two scattered light signals.

In one embodiment, the diluent maintains the original morphology of red blood cells and platelets in the blood sample.

In one embodiment, the at least two scattered light signals may include at least two of an axial light loss, a forward scattered light signal, a mid-angle scattered light signal, a high-angle scattered light signal, a side scattered light signal, and a backward scattered light signal.

In one embodiment, the scattering angles of the axial light loss, the forward scattered light signal, the medium-angle scattered light signal, the high-angle scattered light signal, the side scattered light signal and the backward scattered light signal are respectively 0-1 degree, 1-10 degrees, 10-20 degrees, 20-70 degrees, 70-110 degrees and 110-160 degrees.

In an embodiment, the at least two scattered light signals may comprise at least one, in particular at least two, of a forward scattered signal, a medium angle scattered signal, a high angle scattered signal. Preferably, the at least two scattered light signals may comprise a forward scattered signal and a medium angle scattered signal or may comprise a forward scattered signal and a high angle scattered signal.

In one embodiment, the light irradiation may be polarized light irradiation, and the at least two scattered light signals include at least two of specific polarization state signals of axial light loss, forward scattered light signals, medium angle scattered light signals, high angle scattered light signals, side scattered light signals, and backward scattered light signals of particles in the sample fluid due to the polarized light irradiation. Illustratively, the at least two scattered light signals may comprise at least one, in particular at least two, polarization-specific signals of a forward scattered signal, a medium angle scattered signal, a high angle scattered signal. Preferably, the at least two scattered light signals may comprise polarization-specific signals of the forward scattered signal and the medium angle scattered signal or may comprise polarization-specific signals of the forward scattered signal and the high angle scattered signal.

In an embodiment, the step of classifying the red blood cells and the platelets in the sample to be tested according to the at least two scattered light signals may include:

generating a two-dimensional or three-dimensional scattergram of particles in the blood sample to be tested according to the at least two scattered light signals;

and classifying the red blood cells and the platelets in the blood sample to be detected based on the two-dimensional or three-dimensional scatter diagram.

In one embodiment, the method may include: and obtaining the detection result of the preset parameters of the red blood cells and the platelets in the blood sample to be detected according to the at least two scattered light signals. Illustratively, the predetermined parameter may include a parameter calculated from at least one of a red blood cell count, a platelet count, a mean volume of red blood cells, a mean volume of platelets, and a width of a distribution of the volume of red blood cells, or a combination thereof.

In one embodiment, the method may further include: and outputting the classification result of the red blood cells and the platelets of the blood sample to be detected and/or the detection result of the preset parameters.

In addition, an embodiment of the present invention further provides a sample analyzer, including:

a sampling device having a pipette with a pipette nozzle and having a driving device for driving the pipette to quantitatively aspirate a blood sample through the pipette nozzle;

the sample liquid preparation device is provided with a reaction tank and a liquid supply part, wherein the reaction tank is used for receiving the blood sample sucked by the sampling device, and the liquid supply part is used for supplying diluent to the reaction tank, so that the blood sample sucked by the sampling device and the diluent supplied by the liquid supply part are mixed in the reaction tank to prepare sample liquid to be tested;

the optical detection device is provided with a light source, a flow chamber and a light collector, wherein particles in a sample liquid to be detected after being treated by a diluent can flow in the flow chamber, the particles in the flow chamber are irradiated by light emitted by the light source to generate at least two scattered light signals, and the light collector is used for collecting the at least two scattered light signals;

the conveying device is used for conveying the sample liquid to be detected after the dilution liquid treatment in the reaction tank to the optical detection device;

a processor communicatively coupled to the sampling device, the sample preparation device, the optical detection device, and the delivery device and configured to:

instructing the sample preparation device to prepare a sample solution to be tested containing a blood sample to be tested and a diluent;

instructing the delivery device to deliver the prepared test sample fluid to the flow cell;

acquiring at least two scattered light signals generated by the particles in the sample liquid to be detected in the flow chamber due to light irradiation from the optical detection device;

In one embodiment, the diluent maintains the original morphology of the red blood cells and the platelets in the blood sample.

Illustratively, the at least two scattered light signals may include at least two of an axial light loss, a forward scattered light signal, a mid-angle scattered light signal, a high-angle scattered light signal, a side scattered light signal, and a backward scattered light signal.

In one embodiment, the light source may be configured as a light source emitting polarized light, and the at least two scattered light signals include at least two of specific polarization state signals of an axial light loss, a forward scattered light signal, a medium angle scattered light signal, a high angle scattered light signal, a side scattered light signal, and a backward scattered light signal of particles in the sample fluid due to the polarized light irradiation.

In an embodiment, the at least two scattered light signals comprise at least one, in particular at least two, polarization-specific signals of a forward scattered signal, a medium angle scattered signal, a high angle scattered signal.

In one embodiment, the processor may be configured to:

In one embodiment, the processor may be configured to: and obtaining the detection result of the preset parameters of the red blood cells and the platelets in the blood sample to be detected according to the at least two scattered light signals. Illustratively, the predetermined parameter may include a parameter calculated from at least one of a red blood cell count, a platelet count, a mean volume of red blood cells, a mean volume of platelets, and a width of a distribution of the volume of red blood cells, or a combination thereof.

In one embodiment, the sample analyzer may further include an output device communicatively connected to the processor for outputting the result of the classification of the red blood cells and platelets and/or the result of the detection of the predetermined parameter of the blood sample to be tested.

Advantageous effects

According to the method for detecting the red blood cells and the platelets in the blood sample and the sample analyzer, accurate classification of RBC and PLT can be achieved, and especially accurate classification of RBC and PLT can be achieved under the environment of common diluent.

Drawings

FIG. 1 is a schematic view showing the construction of an optical detection device of a blood cell analyzer;

FIG. 2 is a schematic diagram of an embodiment of a sample optical detection device;

FIG. 3 is a schematic diagram for explaining the structures of the first forward scattered light signal collecting element and the second forward scattered light signal collecting element;

FIG. 4 is a schematic view for explaining the structure of a reflection light blocking member;

FIG. 5 is a schematic diagram of a sample optical inspection device including a lens assembly in one embodiment;

FIG. 6 is a schematic diagram of a configuration for a lens assembly;

FIG. 7 is a schematic diagram of an exemplary optical inspection apparatus including a stop-stop diaphragm;

FIG. 8 is a schematic diagram of an exemplary optical sample detection device including a third forward scattered light signal collection assembly according to an embodiment;

FIG. 9 is a schematic diagram for explaining the structure of a third forward scattered light signal collecting module;

FIG. 10 is a schematic diagram of an exemplary optical inspection device including a side scatter signal collection assembly and a side fluorescence signal collection assembly;

FIG. 11 is a schematic diagram for explaining the structure of a side scattered light signal collecting module and a side fluorescent signal collecting module;

FIG. 12 is a schematic diagram of an exemplary optical inspection device including a light source shaping assembly according to an embodiment;

FIG. 13 is a schematic structural diagram of an optical sample detection device according to another embodiment;

FIG. 14 is a histogram of RBC and PLT measurements using an impedance method;

fig. 15A is an RBC histogram obtained by detecting RBCs and PLTs of a normal sample by an impedance method;

FIG. 15B is a PLT histogram of RBC and PLT of normal samples detected by impedance method;

FIG. 16 is a PLT histogram of RBC and PLT using impedance method to detect abnormal samples;

FIG. 17 is a schematic diagram of a sample analyzer according to one embodiment;

FIG. 18 is a schematic flow chart of a sample detection method for detecting red blood cells and platelets in a blood sample according to an exemplary embodiment;

FIG. 19 is a graph illustrating scattering angles of various scattered light signals according to one embodiment;

FIG. 20 is a scattergram of forward scattered light signals (FSC) and medium angle scattered light signals (MAS) obtained by the sample detection method according to one embodiment;

fig. 21 is a scatter diagram of forward scattered light signals (FSC) and high angle scattered light signals (WAS) obtained by the sample detection method according to an embodiment.

Modes for carrying out the invention

The present invention will be described in further detail with reference to the following detailed description and accompanying drawings. Wherein like elements in different embodiments are numbered with like associated elements. In the following description, numerous details are set forth in order to provide a better understanding of the present application. However, those skilled in the art will readily recognize that some of the features may be omitted or replaced with other elements, materials, methods in different instances. In some instances, certain operations related to the present application have not been shown or described in detail in order to avoid obscuring the core of the present application from excessive description, and it is not necessary for those skilled in the art to describe these operations in detail, so that they may be fully understood from the description in the specification and the general knowledge in the art.

Furthermore, the features, operations, or characteristics described in the specification may be combined in any suitable manner to form various embodiments. Also, the various steps or actions in the method descriptions may be transposed or transposed in order, as will be apparent to one of ordinary skill in the art. Thus, the various sequences in the specification and drawings are for the purpose of describing certain embodiments only and are not intended to imply a required sequence unless otherwise indicated where such sequence must be followed.

The numbering of the components as such, e.g., "first", "second", etc., is used herein only to distinguish the objects as described, and does not have any sequential or technical meaning. The term "connected" and "coupled" when used in this application, unless otherwise indicated, includes both direct and indirect connections (couplings).

Referring to fig. 2, an embodiment of the present invention provides an optical sample detection device, which includes a light source 1, a flow cell 6, a first forward scattered light signal collection assembly 20 and a second forward scattered light signal collection assembly 30, which are described in detail below.

The flow cell 6 is used for allowing the cells in the sample to be tested to pass through one by one. For example, after the blood cells in the sample to be tested have been lysed or stained with reagents, the blood cells are caused to line up one after the other from flow chamber 6 using sheath flow techniques. The Y-axis direction in the figure is a direction in which blood cells in a sample move, and the Y-axis direction in the figure is a direction perpendicular to the plane of the drawing.

The light source 1 is used to illuminate cells passing through the flow cell 6. In one embodiment, the light source 1 is a laser, such as a he — ne laser or a semiconductor laser. When light from the light source 1 is applied to the cells in the flow cell 6, it is scattered to the surroundings. In one embodiment of the present invention, the first forward scattered light signal collection assembly 20 and the second forward scattered light signal collection assembly 30 are used to collect forward scattered light of a first angular range and forward scattered light of a second angular range, respectively, as described in detail below.

The first forward scattered light signal collection assembly 20 is configured to collect forward scattered light of a first angular range, which is light generated by the light source 1 illuminating the cells passing through the flow cell 6 and reflected. Referring to fig. 3, in an embodiment, the first forward scattered light signal collecting assembly 20 includes a first angle limiting diaphragm 21, a stray light blocking diaphragm 22 and a photodetector 23, which are sequentially disposed; the first angle limiting diaphragm is used for limiting the reflected forward scattered light in a first angle range and converging the forward scattered light on the stray light blocking diaphragm 22, the stray light blocking diaphragm 22 is used for shielding stray light of the forward scattered light in the first angle range, and the photoelectric detector 23 is used for converting the collected forward scattered light in the first angle range into an electric signal. In an embodiment, the first angular range is a low angular range, for example the first angular range is 0 to 10 degrees or 1 to 10 degrees.

The second forward scattered light signal collection assembly 30 is used to collect forward scattered light over a second range of angles, which is the light generated by the light source 1 illuminating the cells passing through the flow cell 6. In one embodiment, the second forward scattered light signal collecting assembly 30 comprises a stray light shielding diaphragm 31 and a photodetector 32, which are sequentially arranged, wherein the stray light shielding diaphragm 31 is used for shielding stray light of the forward scattered light in the second angular range, and the photodetector 32 is used for converting the collected forward scattered light in the second angular range into an electrical signal. In one embodiment, the second angular range is a medium angular range, for example, the second angular range is 10 degrees to 20 degrees. In an embodiment, the first and second angular ranges are continuous ranges.

The first forward scattered light signal collecting component 20 collects the forward scattered light of the first angular range after reflection, and the second forward scattered light signal collecting component 30 directly collects or collects the refracted or transmitted forward scattered light, so that the longitudinal dimension (the Z-axis direction in the figure) of the light path can be compressed through the design of the optical structure, and the designed sample optical detection device is miniaturized.

In one embodiment of the present invention, the separation of the forward scattered light over the first and second angular ranges is achieved by a reflective baffle assembly, one reflected to the first forward scattered light signal collection assembly 20 and one allowed to pass to the second forward scattered light signal collection assembly 30, as described in more detail below.

The sample optical detection apparatus in one embodiment further includes a reflective light-blocking assembly 10, wherein the reflective light-blocking assembly 10 is disposed on the optical path of the forward scattered light generated by the light source 1 irradiating the cells passing through the flow cell 6, and is configured to reflect the forward scattered light of the first angle range to the first forward scattered light signal collection assembly 20, and allow the forward scattered light of the second angle range to be directly incident to the second forward scattered light signal collection assembly 30. Referring to fig. 4 (a), the light reflecting and blocking assembly 10 can be implemented in many ways, for example, in one embodiment, the light reflecting and blocking assembly 10 includes a reflector 11 for reflecting the forward scattered light of the first angular range to the first forward scattered light signal collecting assembly 20. In one embodiment, the reflector 11 may be elliptical, i.e. the reflector 11 is an elliptical aperture reflector. The angle of the mirror 11 may be 45 degrees, and the mirror surface is parallel to the Y axis in the drawing, but the angle of the mirror 11 may also be adjusted according to the spatial layout of the light path, and is not limited to only 45 degrees. Referring to fig. 4 (b), in an embodiment, the reflective light-blocking assembly 10 includes a light-blocking strip 12 for blocking stray light except for the forward scattered light in the second angle range. In one embodiment, the reflector 11 is disposed on the light-shielding strip 12, for example, when the reflector 11 is elliptical, the reflector 11 is disposed on the light-shielding strip 12 in a manner that the major axis coincides with the light-shielding strip 12, as shown in fig. 4 (c). The light shielding strip 12 plays a role of a support of the reflector 11, and shields stray light except forward scattered light in the second angle range, so that the signal quality of the forward scattered light in the second angle range can be effectively ensured.

In order to further improve the signal quality of the forward scattered light in the first angle range and the second angle range, referring to fig. 5, in an embodiment, the sample optical detection apparatus may further include a lens assembly 13, where the lens assembly 13 is disposed on the optical path of the forward scattered light generated by the light source 1 irradiating the cells passing through the flow cell 6, and is used for collecting the forward scattered light in the first angle range and the second angle range and emitting the collected forward scattered light to the reflection light blocking assembly 10. For example, in one embodiment, lens assembly 13 includes one aspheric lens and one spherical lens, or a plurality of aspheric lenses, or a plurality of spherical lenses, or an aspheric lens and a plurality of spherical lenses. Referring to fig. 6, an example of an embodiment in which the lens assembly 13 may be formed by two

spherical lenses

14 and 15 is shown. In an embodiment when the lens assembly 13 comprises a spherical lens, the effective data aperture of the spherical lens closest to the flow cell 6 is at least 0.34. The introduction of the lens assembly 13 can ensure that system aberrations are effectively corrected, and that signal spots can be effectively collected by the two measurement channels of the first forward scattered light signal collection assembly 20 and the second forward scattered light signal collection assembly 30. And the signal quality of the forward scattered light in the first angle range can be effectively ensured through the combination of the lens assembly 13 and the reflective light blocking assembly 10.

Referring to fig. 7, in order to further improve the signal quality of the forward scattered light in the first angle range and the second angle range, in an embodiment, the sample optical detection apparatus may further include a straight stop 16 disposed between the lens assembly 13 and the light reflecting and blocking assembly 10. The stop 16 may have a number of functions, such as the ability of the stop 16 to block the direct angle, i.e., 0 degrees, of the forward scattered light exiting the lens assembly 13, which may prevent signal saturation, and/or the ability to limit the forward scattered light exiting the lens assembly 13 to a first range of angles and a second range of angles.

While the above is some description of the sample optical detection device enabling collection of forward scattered light for the first and second angular ranges, in some embodiments the sample optical detection device is also capable of enabling collection of forward scattered light for a third angular range. For example, in one embodiment, the third angular range is a high angular range, such as a third angular range of 20 degrees to 70 degrees. In some embodiments the first, second and third angular ranges are continuous ranges. The following describes in detail how the sample optical detection device achieves the collection of forward scattered light in a third angular range.

Considering that the optical path space behind the flow cell 6 in the sample optical detection device is relatively tight, for example, it is provided with the first forward scattered light signal collection component 20 and the second forward scattered light signal collection component 30, even the lens component 13, etc., if the forward scattered light is directly collected by the photodetector, it cannot be guaranteed that the forward scattered light of the third angular range (e.g., the high angular range) can be completely collected, so the inventors consider the optical path structure for refraction to turn the forward scattered light of the third angular range (e.g., the high angular range), so that the forward scattered light of the third angular range (e.g., the high angular range) can be effectively collected.

Therefore, referring to fig. 8, in one embodiment, the optical detecting apparatus further includes a third forward scattered light signal collecting component 40 for collecting forward scattered light of a third angular range, the forward scattered light being light generated by the light source 1 irradiating the cells passing through the flow cell 6 and having undergone at least one reflection. Referring to fig. 9, in an embodiment, the third forward scattered light signal collecting assembly 40 includes a reflector 41, a third angular range aperture stop 42 and a photodetector 43, which are sequentially disposed; the reflector 41 is used for reflecting the forward scattered light of the third angular range generated by the cells irradiated by the light source 1 through the flow cell 6 to the third angular range aperture stop 42, the third angular range aperture stop 42 is used for limiting the forward scattered light in the third angular range, and the photodetector 43 is used for converting the collected forward scattered light of the third angular range into an electric signal. It should be noted that the optical detection device for sample shown in fig. 9 is only an example shown for illustrating the third forward scattered light signal collecting element 40, and this does not mean that the optical detection device for sample including the third forward scattered light signal collecting element 40 can only have the structure shown in fig. 9.

While the above is some description of the collection of forward scattered light by the sample optical detection device, in some embodiments, the sample optical detection device of the present invention can also achieve the collection of side light, such as side scattered light and/or side fluorescent light, as described in detail below in conjunction with fig. 10 and 11.

The optical sample detection device of an embodiment further includes a side scattered light signal collection unit 50 for collecting side scattered light generated by the light source 1 irradiating the cells passing through the flow cell 6. In one embodiment, the laterally scattered light signal collecting assembly 50 includes a lateral aperture stop 51 and a photodetector 52, which are sequentially disposed, the lateral aperture stop 51 is used for processing the laterally scattered light, and the photodetector 52 receives the laterally scattered light processed by the lateral aperture stop 51 and converts the laterally scattered light into an electrical signal.

The optical sample detection device of an embodiment further includes a lateral fluorescence signal collecting assembly 60 for collecting lateral fluorescence generated by the light source 1 irradiating the cells passing through the flow cell 6. In one embodiment, the lateral fluorescence signal collecting assembly 60 includes a fluorescence aperture stop 61, a filter 62 and a photodetector 63, which are sequentially disposed, where the fluorescence aperture stop 61 is configured to process lateral fluorescence, the processed lateral fluorescence passes through the filter 62 and reaches the photodetector 63, and the photodetector 63 is configured to convert the lateral fluorescence into an electrical signal.

In one embodiment, a collecting lens 7 may be disposed in the path of the side light in the flow cell 6 to collect the side light (including the side scattered light and the side fluorescent light), and the side scattered light may be reflected by a dichroic mirror 8 to the side scattered light signal collecting component 50, for example, the dichroic mirror 8 focuses the side scattered light onto the side aperture stop 51 and then enters the photodetector 52; the lateral fluorescence enters the lateral fluorescence signal collecting assembly 60 through the dichroic mirror 8, for example, the lateral fluorescence passes through the dichroic mirror 8, is focused on the fluorescent aperture stop 61, and then passes through the optical filter 62 to reach the photodetector 63.

Some of the above descriptions of the collection of side light by the sample optical detection device, the side scattered light and the side fluorescent light may be 70 degrees to 110 degrees in one embodiment.

It should be noted that fig. 10 and 11 are examples of the sample optical detection device including both the side scattered light signal collection assembly 50 and the side fluorescent signal collection assembly 60, and in some embodiments, the sample optical detection device may include only one of the side scattered light signal collection assembly 50 and the side fluorescent signal collection assembly 60.

While the forward scattered light, the side scattered light and the side fluorescent light generated by the light source 1 irradiating the cells passing through the flow cell 6 are collected by the corresponding measurement channels, in some embodiments, the light path from the light source 1 to the flow cell 6 may also be provided with some devices for improving the optical signal, for example, referring to fig. 12, in an embodiment, the sample optical detection apparatus may further include a light source shaping component 9 for collimating and focusing the light beam emitted from the light source 1 on the cells passing through the flow cell 6. Referring to fig. 13, in an embodiment, the light source shaping component 9 includes a collimating lens 2, an optical isolator 3, a first cylindrical mirror 4 and a second cylindrical mirror 5, which are sequentially disposed; the collimating lens 2 is used for collimating light beams emitted by the light source 1, and the optical isolator 3 is used for inhibiting feedback light and preventing reflected light of a rear optical device from entering the light source 1 and influencing the light source 1; the first cylindrical mirror 4 is used to converge the light beam at the center of the flow cell in the direction in which the cell passes, for example, the first cylindrical mirror 4 converges the light beam at the center of the flow cell in the Y-axis direction in the figure; the second cylindrical mirror 5 is used to converge the light beam in a direction perpendicular to the direction in which the cell passes, for example, the second cylindrical mirror 5 is used to converge the light beam in the X-axis direction in the drawing, so that the scattered light is all irradiated into the straight stop 16. Those skilled in the art will appreciate that components in the source shaping assembly 9 are not necessary in some cases, such as the optical isolator 3 and the second cylindrical mirror 5.

In summary, the five measurement channels in the sample optical detection apparatus are described separately, and in the specific implementation, several of the five measurement channels may be introduced according to the requirement. For example, FIG. 13 above is a schematic diagram of a five measurement channel sample optical detection device. The operation and principle of these five measurement channels will be described below with reference to fig. 13 as an example.

The direction of flow of the sample cells (i.e., the direction through the flow cell 6) in the figure is the Y-axis direction, i.e., the direction perpendicular to the plane of the paper. Light emitted from a light source 1 is changed into an equal beam by a collimator lens 2, passes through an optical isolator 3, and then is processed by a first cylindrical mirror 4 so that the beam converges at the center of the flow cell in the direction in which the sample cell flows, i.e., the Y-axis direction, to irradiate the cell passing through the flow cell 6. The optical isolator 3 mainly has the main function of inhibiting feedback light which mainly comes from the reflection of each optical component behind the optical isolator 3 from entering the light source 1, preventing the output power of the light source 1 from fluctuating and ensuring the stability of an optical baseline.

The second cylindrical lens 5 and the

spherical lenses

14 and 15 can make the light beam converge in the X-axis direction, for example, by designing the second cylindrical lens 5 and the

spherical lenses

14 and 15, the size of the light beam in the X-axis direction is compressed to be smaller than the transverse size of the straight bar of the straight stop 16, so as to ensure that all the direct light can be effectively blocked by the straight stop 16, and prevent signal saturation; note that the bar of the bar stop 16 is a bar in the Y-axis direction, that is, the axial direction of the bar is the Y-axis direction, and the lateral direction is the X-axis direction.

When the processed blood cells pass through the flow cell 6 one by one, the light beam of the light source 1 finally irradiates the cells passing through the flow cell 6, and forward scattered light in a first angle range (hereinafter, the forward scattered light at a low angle is not exemplified), forward scattered light in a second angle range (hereinafter, the forward scattered light at a medium angle is not exemplified), forward scattered light in a third angle range (hereinafter, the forward scattered light at a high angle is not exemplified), side scattered light, and side fluorescence are generated:

the low-angle forward scattered light and the medium-angle forward scattered light are collected by the

spherical lenses

14 and 15, and then the low-angle forward scattered light is reflected by the reflective light blocking assembly 10 (e.g., an elliptical aperture mirror thereof) to the first angle limiting diaphragm 21 (e.g., a low-angle limiting diaphragm) to converge at the stray light blocking diaphragm 22, and then enters the photodetector 23; the forward scattered light of the medium angle is effectively shielded by the reflection light blocking assembly 10 (for example, the light blocking strip 12 is arranged along the plane direction composed of the X axis and the Z axis, the effective shielding direction is the X axis direction), and then the stray light of the forward scattered light of the medium angle is converged to the stray light blocking diaphragm 31 and enters the photodetector 32;

the high angle forward scattered light is reflected by the mirror 41 to the third angular range aperture stop 42 (e.g., high angle aperture stop) and then enters the photodetector 43;

the side scattered light and the side fluorescent light are simultaneously collected by the collecting lens 7 and spatially separated while passing through the dichroic mirror 8, wherein the side scattered light is reflected and focused on the side aperture stop 51 through the front surface of the dichroic mirror 8 and then enters the photodetector 52; the side fluorescent light passes through the dichroic mirror 8, is focused on the fluorescent aperture stop 61, and then passes through the optical filter 62 to reach the photodetector 63.

The above are some descriptions of the sample optical detection apparatus of the present invention. The lens assembly is used for collecting the low-angle forward scattered light and the medium-angle forward scattered light in the forward scattered light, and the reflection light blocking assembly is used for carrying out space separation on the low-angle forward scattered light and the medium-angle forward scattered light, so that the signal quality of the low-angle forward scattered light and the medium-angle forward scattered light is ensured, the longitudinal size of an optical structure is reduced, and the miniaturization is facilitated; in a compact light path structure of forward scattered light, the invention collects high-angle forward scattered light by using the reflector, which can replace the traditional classification of eosinophil by side scattered light signals; the invention adds a middle-angle forward scattering light measuring channel and a high-angle forward scattering light measuring channel on the low-angle forward scattering light measuring channel, and can classify and count some cells through the three measuring channels, for example, platelets can be classified from red blood cells through the three measuring channels under the condition of not using a fluorescent reagent or a sphering reagent; the invention utilizes the collecting lens arranged on the lateral light path of the flowing chamber to perform depolarization and fluorescence signal collection on the lateral light, and separates the depolarized lateral scattered light and the fluorescence signal through the wavelength selection of the dichroic mirror. The present invention realizes a light path structure for receiving scattered light from 0 degree or 1 degree to 110 degrees, and no lost angle occurs in the middle.

The sample detection method and the sample analyzer of the present invention are described in detail next with reference to fig. 14 to 21.

Currently, impedance methods or fluorescence optical methods are generally used in blood analyzers to measure RBCs and PLTs in blood samples.

The impedance method is based on the coulter principle, so that a diluted blood sample passes through a small hole, constant current sources are applied to two sides of the small hole, and each cell passing through the small hole causes the electrical impedance of liquid in the small hole to change, thereby generating electric pulses. The corresponding electrical pulses are acquired with amplitudes representing the volume of the cells, thereby generating a RBC and PLT histogram, which is a one-dimensional information, i.e., only the volume information of the cells, as shown in fig. 14.

In fig. 14, the PLT histogram is on the left of the dotted line, and the RBC histogram is on the right of the dotted line. Since the RBC particle diameter is about 3 times the PLT and the number of RBC particles is about 30 times the PLT, the PLT histogram is to the left of the RBC histogram and the area enclosed by the horizontal axis is much smaller than that of RBC. That is, the dotted line in fig. 14 indicates the boundary between the RBC histogram and the PLT histogram. Performing normalization processing on the RBC histogram and the PLT histogram respectively, and drawing out the RBC histogram and the PLT histogram separately to form the RBC histogram and the PLT histogram which are common on a blood cell analyzer, as shown in fig. 15A and 15B, wherein fig. 15A is the RBC histogram obtained by detecting the RBC and the PLT of a normal sample by using an impedance method; fig. 15B is a PLT histogram of RBC and PLT of normal samples detected by impedance method.

In the RBC and PLT histograms of normal samples, there is an obvious boundary between the RBC peak and the PLT peak, and the valley on the right side of the main peak of the PLT particle in the PLT histogram is usually used as the boundary between the PLT and the RBC, and then the two histograms are analyzed respectively to obtain the measurement parameters related to the PLT and the RBC. However, for some abnormal samples, such as low-value PLT samples, the boundary between the PLT histogram and the RBC histogram often has no obvious boundary, as shown in fig. 16, the low-value PLT histogram is jagged, so that the algorithm cannot accurately cut the PLT and RBC histograms, and further cannot obtain an accurate PLT measurement result. The accuracy of low value PLTs is a routine indicator of blood of major clinical concern, which is a major drawback of impedance measurements.

Fluorescence optics can overcome this drawback. Fluorescence methods are based on flow cytometry. The diluted and stained sample is passed sequentially through the optical detection zone under the extrusion of a sheath fluid. Each cell is irradiated by an excitation light source, and a forward scattering signal (representing the cell volume) and a fluorescence signal (representing the content of nucleic acid in the cell) are obtained in an optical system, so that a two-dimensional scatter diagram is generated, and the RBC and the PLT are divided and calculated. Compared with the one-dimensional histogram obtained by the impedance method, the two-dimensional scatter diagram of the optical method has one more dimension of information, so that the PLT and the RBC can be accurately divided on the two-dimensional scatter diagram. However, in the fluorescence method, a fluorescent dye is required to stain cells, and special diluent is required to perform sphericization treatment on the cells, so that the use cost is high, and the popularization of the optical method in clinic is not facilitated.

In order to solve the problem, the invention provides a sample detection method and a sample analyzer for detecting red blood cells and platelets in a blood sample by using only scattered light information, wherein the sample detection method and the sample analyzer can realize accurate classification of RBC and PLT in a common diluent environment, and particularly can realize accurate classification of RBC and PLT in an abnormal sample in a common diluent environment.

First, a sample analyzer according to the present invention will be described in detail with reference to fig. 17.

As shown in fig. 17, the sample analyzer 100 includes at least a sampling device (not shown), a sample liquid preparation device 110, an optical detection device 120, a transport device 130, and a processor 140.

The sampling device has a pipette with a pipette nozzle and has a drive device for driving the pipette to aspirate a blood sample quantitatively through the pipette nozzle. Further, the sampling device is driven by its driving means and moved to the reaction cell 111 of the sample liquid preparation means 110 after sucking the blood sample, and the sucked blood sample is injected into the reaction cell 111.

The sample liquid preparation device 110 has at least one reaction cell 111 and a liquid supply portion (not shown), wherein the reaction cell 111 is used for receiving the blood sample sucked by the sampling device, and the liquid supply portion supplies a diluent to the reaction cell 111, so that the blood sample sucked by the sampling device and the diluent supplied by the liquid supply portion react in the reaction cell to prepare a blood sample to be detected. For example, the liquid supply portion may be used to inject an appropriate diluent into the reaction cell to process particles in the blood sample, so as to prepare the blood sample to be detected for subsequent detection. Wherein the diluent is a common diluent necessary for a blood cell analyzer, so as to maintain the original forms of the red blood cells and the platelets in the blood sample, and a special diluent for spheroidizing RBC and PLT is not required. For example, the diluent may include sodium chloride, phosphate buffer, and preservative, and the diluent is not limited to a specific one, and may be selected according to the need, and will not be described herein.

The optical detection device 120 has a light source 121, a flow cell 122, and

light collectors

123, 124. The light source 121 may emit natural light or light of a specific wavelength band, and is not limited to a certain one. Alternatively, the light source 121 may be a polarized light source to emit polarized light of a specific polarization state. The flow cell 122 has an orifice 1221, and particles of the sample liquid to be measured after being treated with the diluting liquid in the sample liquid preparation apparatus 110 can flow in the flow cell 122 and pass through the orifice 1221 one by one. Light emitted by light source 121 illuminates particles in the flow cell 122 to produce optical signal information. The

light collectors

123, 124 are used to collect the optical signal information. The optical signal information may include at least two of an axial light loss, a forward scattered light signal, a medium angle scattered light signal, a high angle scattered light signal, a side scattered light signal, and a backward scattered light signal, and when irradiation with polarized light is selected, the optical signal information includes at least two of specific polarization state signals of the axial light loss, the forward scattered light signal, the medium angle scattered light signal, the high angle scattered light signal, the side scattered light signal, and the backward scattered light signal. That is, the optical detection device 120 includes at least two of an axial light loss collector, a forward scattered light signal collector, a middle angle scattered light signal collector, a high angle scattered light signal collector, a side scattered light signal collector, and a backward scattered light signal collector.

In one embodiment, the light collector is configured as a photodetector, such as a photodiode or photomultiplier tube. Specifically, as shown in fig. 17, forward scattered light emitted from blood cells flowing in the flow cell 122 is received by the photodiode (forward scattered light collector) 123 through the condenser 126 and the pinhole 127, and side scattered light is received by the photomultiplier (side scattered light collector) 124 through the condenser 126, the dichroic mirror 128, the optical film 129, and the pinhole 127. The optical signals output from the

respective light collectors

123 and 124 are subjected to analog signal processing such as amplification and waveform processing by an amplifier 150, and then are transmitted to a processor 140.

In the invention, the scattering angles of the axial light loss, the forward scattered light signal, the medium-angle scattered light signal, the high-angle scattered light signal, the side scattered light signal and the backward scattered light signal are respectively 0-1 degree, 1-10 degrees, 10-20 degrees, 20-70 degrees, 70-110 degrees and 110-160 degrees. The axial light loss light collector, the forward scattered light signal light collector, the middle angle scattered light signal light collector, the high angle scattered light signal light collector and the side scattered light signal light collector are respectively configured to receive scattered light signals of the scattering angles.

Preferably, the optical detection device 120 at least comprises at least one, especially at least two, of a forward scattered light signal collector, a medium angle scattered light signal collector and a high angle scattered light signal collector to receive at least one, especially at least two, of a forward scattered signal, a medium angle scattered signal and a high angle scattered signal or at least one, especially at least two, of specific polarization state signals of the forward scattered signal, the medium angle scattered signal and the high angle scattered signal, so as to improve the accuracy of the detection result of the red blood cells and the platelets. In particular, the optical detection device 120 includes a forward scattered light signal collector and a medium angle scattered light signal collector or includes a forward scattered light signal collector and a high angle scattered light signal collector.

The transportation device 130 is used for transporting the blood sample to be tested, which is processed by the dilution liquid in the reaction cell 111, to the optical detection device 120.

The processor 140 is communicatively connected to the sampling device, the sample preparation device 110, the optical detection device 120, and the transport device 130 and is configured to acquire the optical signal information from the optical detection device 120 and process the optical signal information to obtain a result of the particle classification count in the blood sample to be tested. The processor 140 may have an a/D converter, not shown, for converting the analog signal provided by the optical detection device 120 into a digital signal. In particular, the processor 140 is configured to implement a sample detection method according to the present invention, which is described in further detail below.

In the embodiment of the present invention, the Processor 140 may be at least one of an Application Specific Integrated Circuit (ASIC), a Digital Signal Processor (DSP), a Digital Signal Processing Device (DSPD), a ProgRAMmable Logic Device (PLD), a Field ProgRAMmable Gate Array (FPGA), a Central Processing Unit (CPU), a controller, a microcontroller, and a microprocessor. It will be appreciated that the electronic devices used to implement the processor functions described above may be other devices, and embodiments of the present invention are not limited in particular.

Next, a detailed description will be given of a specific method and principle of detecting red blood cells and platelets in a blood sample by the above-described blood sample analyzer 100 with reference to fig. 18 to 21.

FIG. 18 is a sample detection method for detecting red blood cells and platelets in a blood sample according to an embodiment. As shown in fig. 18, a sample detection method 200 for detecting red blood cells and platelets in a blood sample includes the steps of:

step S210, preparing a sample solution to be detected containing a blood sample to be detected and a diluent.

For example, in this step, a sample liquid to be tested is prepared, for example, in a sample preparation device 110 of the sample analyzer 100, the sample liquid preparation device 110 has at least one reaction cell 111 and has a liquid supply portion (not shown), under the control of the processor 140, the sampling device is driven by its driving device and moves to the reaction cell 111 of the sample liquid preparation device 110 after sucking the blood sample, and then the liquid supply portion supplies a diluent to the reaction cell 111, so that the blood sample sucked by the sampling device reacts with the diluent supplied by the liquid supply portion in the reaction cell to prepare a blood sample to be tested. On one hand, the diluent provides proper pH, conductivity and osmotic pressure for a blood sample to be detected, so that the complete form of cells is ensured, and hemolysis is avoided; in addition, the diluent is also used for cleaning substances remained in the sample detected last time, so that the cleanness of a sampling needle, a pipeline and a flow chamber of a sample analyzer is ensured, and cross contamination is prevented; on the other hand, the diluent is filled in the middle of the pipeline during the temporary stop of the sample analyzer, so that foreign matters such as external dust are prevented from entering the analyzer, and faults are prevented. Wherein the diluent is a common diluent necessary for a blood cell analyzer, so as to maintain the original forms of the red blood cells and the platelets in the blood sample, rather than a special diluent for spheroidizing RBC and PLT.

In an embodiment of the present invention, the diluent may include sodium chloride, a phosphate buffer, a preservative, and other components, and the diluent is not limited to a certain one and may be selected according to needs, which is not described herein again.

Step S220, performing light irradiation on the sample liquid to be detected in the optical detection area.

For example, in this step, under the control of the processor 140, the transportation device 130 of the sample analyzer 100 transports the blood sample to be tested, which is processed by the dilution solution in the reaction cell 111, into the flow cell 122 of the optical detection device 120, particles of the sample solution to be tested, which is processed by the dilution solution, can flow in the flow cell 122 and pass through the orifice 1221 one by one, and the light emitted by the light source 121 irradiates the particles in the flow cell 122 to generate optical signal information.

Step S230, collecting at least two kinds of scattered light signals generated by the particles in the sample liquid to be measured due to light irradiation, that is, the scattered light signals at a specific scattering angle.

For example, in this step, under the control of the processor 140, the optical detection device 120 transmits the scattered light signal output therefrom to the processor 140, so that the processor 140 processes the scattered light signal. In an embodiment of the present invention, the scattered light signals may include at least two of axial light loss, forward scattered light signals, medium angle scattered light signals, high angle scattered light signals, side scattered light signals, and backward scattered light signals. Preferably, the scattered light signal comprises at least one, in particular at least two, of a forward scatter signal, a medium scatter signal, a high scatter signal. More preferably, the at least two scattered light signals comprise a forward scattered signal and a medium angle scattered signal or comprise a forward scattered signal and a high angle scattered signal.

In the embodiment of the present invention, the scattering angle refers to an angle formed by a vertex, a first side, a second side, and two sides, the vertex is an angle formed by a center of a region where the sample flow and the excitation beam overlap in the flow cell, the first side is an angle formed by a propagation direction of the excitation beam, and the second side is an angle formed by a propagation direction of scattered light emitted by particles at the vertex. Without going to the contrary, the scattering angles refer to this explanation.

Here, the present invention defines the scattering angle of the scattered light signal as follows, as shown in fig. 19, in which: the scattering angle of the axial light loss is: 0-1 degree; the scattering angle of the forward scattering optical signal is as follows: 1-10 degrees; the scattering angle of the medium-angle scattered light signal is as follows: 10-20 degrees; the scattering angle of the high-angle scattered light signal is as follows: 20-70 degrees; the scattering angle of the side scattering optical signal is as follows: 70-110 degrees; and the scattering angle of the backscattered light signal is: 110-160 degrees.

In an embodiment of the present invention, any two or all of the forward scattering signal, the medium scattering signal and the high scattering signal are collected for the subsequent classification of the red blood cells and the platelets in the blood sample, and at least two of the three scattering signals are selected to more effectively and accurately classify the red blood cells and the platelets. Specifically, for example, a scatter diagram of red blood cells and platelets obtained by selecting at least two of the forward scatter signal, the medium scatter signal, and the high scatter signal has a clearer boundary between red blood cells and platelets, so that the classification result is more accurate.

Of course, the selection of the scattered light signal is not limited to the above example, and may be selected according to actual needs.

The light irradiation can be natural light or light with a specific wave band, and polarized light irradiation with a specific polarization state can also be selected. When the sample liquid to be detected is irradiated by polarized light, the scattered light signals are specific polarization state signals of axial light loss, forward scattered light signals, middle-angle scattered light signals, high-angle scattered light signals, side scattered light signals and backward scattered light signals generated by the particles in the sample liquid due to the irradiation of the polarized light.

Similarly, in an embodiment of the present invention, any two or all of the polarization-specific signals of the forward scatter signal, the medium angle scatter signal, and the high angle scatter signal are collected for subsequent classification of red blood cells and platelets in the blood sample for more efficient and accurate classification of red blood cells and platelets.

Step S240, classifying the red blood cells and the platelets in the blood sample to be tested according to the at least two scattered light signals.

For example, the processor 140 receives the scattered light signal from the optical detection device 120 and processes the scattered light signal to obtain a classification result of red blood cells and platelets in the blood sample to be tested.

Preferably, in this step S240, the step of classifying the red blood cells and the platelets may include: generating a two-dimensional or three-dimensional scattergram of particles in the blood sample from the at least two scattered light signals; classifying red blood cells and platelets in the blood sample based on the two-dimensional or three-dimensional scattergram.

In an embodiment of the present invention, for example, when the PLT value of the sample fluid to be measured is less than 30, the PLT histogram appears jagged in the right boundary region, as shown in fig. 16, and it is difficult to demarcate a boundary line separated from the RBC region. However, the detection method of the present invention can generate a scattergram as shown in fig. 20 and 21. Fig. 20 is a scatter diagram of a Forward Scattering (FSC) optical signal and a medium-angle scattering (MAS) optical signal. Fig. 21 is a scatter plot of Forward Scattered (FSC) light signals versus high angle scattered (WAS) light signals. It can be seen from fig. 20 and 21 that there is a distinct boundary between the PLT and RBC clusters, as indicated by the direction of the dashed lines in the figures. Therefore, PLTs and RBC clusters that cannot be distinguished in the impedance method can be distinguished in the light scattering scattergram, resulting in accurate RBC and PLT measurements.

In addition to the classification of the red blood cells and the platelets, the detection result of the predetermined parameter of the red blood cells and the platelets can be obtained from the at least two scattered light signals. The predetermined parameter may include at least one of a red blood cell count, a platelet count, a mean volume of red blood cells MCV, a mean volume of platelets MPV, and a red blood cell volume distribution width RDW, and other parameters calculated by combining the above parameters.

Further, the method 200 may further include outputting a result of the differential counting of the red blood cells and the platelets of the blood sample to be tested and/or a result of the detection of the predetermined parameter.

The method 200 for detecting red blood cells and platelets in a blood sample and the corresponding sample analyzer according to the present invention can realize accurate classification of RBC and PLT, especially in a common diluent environment. In the case of a normal diluent, the red blood cells and platelets in the blood sample retain their original morphology, e.g., the red blood cells retain a biconcave pie shape with thicker edges and thinner centers, and are therefore difficult to detect and distinguish. This drawback can be compensated by using at least two scattered lights to collect more cell features, with only the scattered light being used to differentiate between red blood cells and platelets in the case of a common diluent.

The present invention has been described in terms of specific examples, which are provided to aid understanding of the invention and are not intended to be limiting. Variations of the above-described embodiments may be made by those skilled in the art, consistent with the principles of the invention.

Claims

An optical sample detection device, comprising:

the flow chamber is used for allowing cells in a sample to be detected to pass through one by one;

a light source for illuminating cells passing through the flow cell;

a first forward scattered light signal collection assembly for collecting forward scattered light of a first angular range, the forward scattered light being light generated by the light source illuminating cells passing through the flow cell and reflected;

a second forward scattered light signal collection assembly for directly collecting forward scattered light of a second angular range, the forward scattered light being light generated by the light source illuminating cells passing through the flow cell.
The optical sample detection device according to claim 1, further comprising a light-reflecting blocking member disposed in an optical path of forward scattered light generated by the light source irradiating the cells passing through the flow cell, for reflecting the forward scattered light of the first angle range to the first forward scattered light signal collecting member and allowing the forward scattered light of the second angle range to be directly incident to the second forward scattered light signal collecting member.
The sample optical assay of claim 2, wherein the reflective light barrier assembly comprises a mirror for reflecting forward scattered light of a first angular range to the first forward scattered light signal collection assembly.
The optical sample detection device as claimed in claim 3, wherein the mirror is elliptical.
The optical detection device as claimed in claim 3 or 4, wherein the reflective light-blocking assembly further comprises a light-blocking strip for blocking stray light except for the forward scattered light of the second angle range; the reflector is arranged on the shading strip.
The optical detection device as claimed in claim 5, wherein the mirror is disposed on the light-shielding strip such that a long axis thereof coincides with the light-shielding strip.
The optical sample detection device as claimed in any one of claims 2 to 6, further comprising a lens assembly disposed on an optical path of the forward scattered light generated by the light source irradiating the cells passing through the flow cell, for collecting the forward scattered light of the first angle range and the second angle range and emitting the forward scattered light to the reflective light blocking assembly.
The sample optical assay device of claim 7, wherein the lens assembly comprises one aspheric lens and one spherical lens, or a plurality of aspheric lenses, or a plurality of spherical lenses, or one aspheric lens and a plurality of spherical lenses.
The sample optical assay device of claim 8, wherein when the lens assembly comprises a spherical lens, an effective data aperture of the spherical lens closest to the flow cell is at least 0.34.
The optical sample detection device as claimed in claim 7, further comprising a stop disposed between the lens assembly and the reflective light blocking assembly for blocking the direct angle of the forward scattered light emitted from the lens assembly and/or limiting the forward scattered light emitted from the lens assembly to the first angle range and the second angle range.
The optical sample detection device according to claim 1, wherein the first forward scattered light signal collection unit comprises a first angle limiting diaphragm, a stray light blocking diaphragm and a photodetector arranged in this order; the first angle limiting diaphragm is used for limiting the reflected forward scattered light in a first angle range and converging the reflected forward scattered light in the stray light blocking diaphragm, the stray light blocking diaphragm is used for shielding stray light of the forward scattered light in the first angle range, and the photoelectric detector is used for converting the collected forward scattered light in the first angle range into an electric signal.
The optical sample detection device as claimed in claim 1, wherein the second forward scattered light signal collection module comprises a stray light stop for shielding stray light of the forward scattered light of the second angular range and a photodetector for converting the collected forward scattered light of the second angular range into an electrical signal, which are arranged in this order.
The optical sample detection device of claim 1, further comprising a third forward scattered light signal collection assembly for collecting forward scattered light of a third angular range, the forward scattered light being light generated by the light source illuminating cells passing through the flow cell and having undergone at least one reflection.
The optical sample detection device as claimed in claim 13, wherein the third forward scattered light signal collection assembly comprises a mirror, a third angular range aperture stop and a photodetector arranged in this order; the reflector is used for reflecting the forward scattering light of a third angle range generated by cells irradiated by the light source through the flow chamber to the third angle range aperture stop, the third angle range aperture stop is used for limiting the forward scattering light in the third angle range, and the photoelectric detector is used for converting the collected forward scattering light in the third angle range into an electric signal.
The sample optical detection device according to any one of claims 1 to 14, wherein the first angular range is a low angular range, and/or the second angular range is a medium angular range, and/or the third angular range is a high angular range.
The sample optical detection device of claim 15, wherein the first angular range and the second angular range are continuous ranges.
The sample optical detection device of claim 15, wherein the first angular range, the second angular range, and the third angular range are continuous ranges.
The sample optical detection device according to claim 15, wherein the first angle range is 0 to 10 degrees or 1 to 10 degrees; and/or, the second angle ranges from 10 degrees to 20 degrees; and/or the third angle range is 20 degrees to 70 degrees.
The optical sample detection device of claim 1, further comprising a light source shaping assembly for collimating and focusing the light beam emitted by the light source onto the cells passing through the flow cell.
The optical detection device according to any one of claims 1 to 19, wherein the light source shaping assembly comprises a collimating lens and a first cylindrical mirror, which are arranged in sequence, the collimating lens is configured to collimate the light beam emitted by the light source, and the first cylindrical mirror is configured to converge the light beam to the center of the flow chamber in the direction in which the cell passes through.
The sample optical detection device according to claim 20, further comprising a second cylindrical mirror disposed on an exit path of the first cylindrical mirror for converging the light beam in a direction perpendicular to a cell passing direction so that scattered light is irradiated into the direct-blocking diaphragm.
The optical sample detection device as claimed in claim 20, further comprising an optical isolator disposed between the collimating lens and the first cylindrical mirror for suppressing the feedback light.
The optical sample detection device according to claim 1, further comprising:

a side scatter signal collection assembly for collecting side scatter light generated by the light source illuminating cells passing through the flow cell; and/or the presence of a gas in the gas,

a lateral fluorescence signal collection assembly for collecting lateral fluorescence generated by the light source illuminating cells passing through the flow cell.
A sample detection method for detecting red blood cells and platelets in a blood sample, the method comprising:

preparing a sample solution to be detected containing a blood sample to be detected and a diluent;

irradiating the sample liquid to be detected with light in an optical detection area;

collecting at least two scattered light signals generated by the particles in the sample liquid to be detected due to light irradiation, wherein at least one of the at least two scattered light signals is selected from a forward scattering signal, a medium-angle scattering signal or a high-angle scattering signal;

and classifying the red blood cells and the platelets in the blood sample to be detected according to the at least two scattered light signals.
The method of claim 24, wherein the diluent maintains the original morphology of red blood cells and platelets in the test blood sample.
The method of claim 24 or 25, wherein the at least two scattered light signals comprise at least two of a forward scattered signal, a medium angle scattered signal, and a high angle scattered signal.
The method of any one of claims 24 to 26, wherein the forward scattered light signal, the mid-angle scattered light signal, and the high-angle scattered light signal have scattering angles of 1 ° -10 °, 10 ° -20 °, and 20 ° -70 °, respectively.
The method of any one of claims 24 to 27, wherein the at least two scattered light signals comprise a forward scattered signal and a medium angle scattered signal or comprise a forward scattered signal and a high angle scattered signal.
The method of claim 24 or 25, wherein the light irradiation is polarized light irradiation and the at least two scattered light signals comprise at least two of forward scattered light signals, medium angle scattered light signals and specific polarization state signals of high angle scattered light signals generated by the particles in the sample fluid due to the polarized light irradiation.
The method of claim 29, wherein the at least two scattered light signals comprise polarization-specific signals for forward scattered signals and mid-angle scattered signals or polarization-specific signals for forward scattered signals and high-angle scattered signals.
The method of any one of claims 24 to 30, wherein the step of classifying the red blood cells and platelets in the test sample from the at least two scattered light signals comprises:

generating a two-dimensional or three-dimensional scattergram of particles in the blood sample to be tested according to the at least two scattered light signals;

and classifying the red blood cells and the platelets in the blood sample to be detected based on the two-dimensional or three-dimensional scatter diagram.
The method according to any one of claims 24 to 31, characterized in that the method comprises: and obtaining the preset parameters of the red blood cells and the platelets in the blood sample to be detected according to the at least two scattered light signals.
The method of claim 32, wherein the predetermined parameters include parameters calculated from at least one of red blood cell count, platelet count, mean volume of red blood cells, mean volume of platelets, and width of distribution of volume of red blood cells, or a combination thereof.
The method of claim 33, further comprising:

and outputting the classification result and/or the preset parameters of the red blood cells and the platelets of the blood sample to be detected.
A sample analyzer, comprising:

a sampling device having a pipette with a pipette nozzle and having a driving device for driving the pipette to quantitatively aspirate a blood sample through the pipette nozzle;

the sample liquid preparation device is provided with a reaction tank and a liquid supply part, wherein the reaction tank is used for receiving the blood sample sucked by the sampling device, and the liquid supply part is used for supplying diluent to the reaction tank, so that the blood sample sucked by the sampling device and the diluent supplied by the liquid supply part are mixed in the reaction tank to prepare sample liquid to be tested;

the optical detection device is provided with a light source, a flow chamber and a light collector, wherein particles in a sample liquid to be detected after being treated by a diluent can flow in the flow chamber, the particles in the flow chamber are irradiated by light emitted by the light source to generate at least two scattered light signals, and the light collector is used for collecting the at least two scattered light signals;

the conveying device is used for conveying the sample liquid to be detected after the dilution liquid treatment in the reaction tank to the optical detection device;

a processor communicatively coupled to the sampling device, the sample preparation device, the optical detection device, and the delivery device and configured to:

instructing the sample preparation device to prepare a sample solution to be tested containing a blood sample to be tested and a diluent;

instructing the delivery device to deliver the prepared test sample fluid to the flow cell;

acquiring at least two scattered light signals generated by the particles in the sample liquid to be detected in the flow chamber due to light irradiation from the optical detection device, wherein at least one of the at least two scattered light signals is selected from a forward scattering signal, a medium-angle scattering signal or a high-angle scattering signal;

and classifying the red blood cells and the platelets in the blood sample to be detected according to the at least two scattered light signals.
The sample analyzer of claim 35, wherein the diluent maintains the original morphology of red blood cells and platelets in the blood sample to be tested.
The sample analyzer of claim 35 or 36 wherein the at least two scattered light signals comprise at least two of forward scattered light signals, medium angle scattered light signals, and high angle scattered light signals.
The sample analyzer of any of claims 35-37 wherein the forward scattered light signal, the mid-angle scattered light signal, and the high-angle scattered light signal have scattering angles of 1 ° -10 °, 10 ° -20 °, and 20 ° -70 °, respectively.
The sample analyzer of any of claims 35 to 38, wherein the at least two scattered light signals comprise a forward scattered signal and a medium angle scattered signal or comprise a forward scattered signal and a high angle scattered signal.
The sample analyzer of any one of claims 35 to 39 wherein the processor is configured to:

generating a two-dimensional or three-dimensional scattergram of particles in the blood sample to be tested according to the at least two scattered light signals;

and classifying the red blood cells and the platelets in the blood sample to be detected based on the two-dimensional or three-dimensional scatter diagram.
The sample analyzer of any one of claims 35 to 40 wherein the processor is configured to: and obtaining the preset parameters of the red blood cells and the platelets in the blood sample to be detected according to the at least two scattered light signals.
The sample analyzer of claim 41 wherein the predetermined parameters include parameters calculated from at least one of red blood cell count, platelet count, mean volume of red blood cells, mean volume of platelets, and width of red blood cell volume distribution, or combinations thereof.
The sample analyzer of claim 42 further comprising an output device communicatively coupled to the processor for outputting the results of the classification of the red blood cells and platelets and/or the predetermined parameters of the blood sample to be tested.