WO2020147255A1 - 样本光学检测装置、样本检测方法及样本分析仪 - Google Patents

样本光学检测装置、样本检测方法及样本分析仪 Download PDF

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Publication number
WO2020147255A1
WO2020147255A1 PCT/CN2019/090756 CN2019090756W WO2020147255A1 WO 2020147255 A1 WO2020147255 A1 WO 2020147255A1 CN 2019090756 W CN2019090756 W CN 2019090756W WO 2020147255 A1 WO2020147255 A1 WO 2020147255A1
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Prior art keywords
scattered light
sample
light
angle
signal
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PCT/CN2019/090756
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English (en)
French (fr)
Inventor
汪东生
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Shenzhen Mindray Bio Medical Electronics Co Ltd
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Shenzhen Mindray Bio Medical Electronics Co Ltd
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Priority to CN201980080985.4A priority Critical patent/CN113196039B/zh
Publication of WO2020147255A1 publication Critical patent/WO2020147255A1/zh
Anticipated expiration legal-status Critical
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/47—Scattering, i.e. diffuse reflection
    • G01N21/49—Scattering, i.e. diffuse reflection within a body or fluid
    • G01N21/53—Scattering, i.e. diffuse reflection within a body or fluid within a flowing fluid, e.g. smoke
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/483—Physical analysis of biological material
    • G01N33/487—Physical analysis of biological material of liquid biological material
    • G01N33/49—Blood

Definitions

  • the invention relates to a sample optical detection device, a sample detection method and a sample analyzer.
  • Most blood cell analyzers use the principle of laser scattering for measurement.
  • the principle is: irradiate the laser on the cell, collect the forward scattered light, side scattered light (90 degree scattered light) and side fluorescence (90 degree scattered light) produced by the cells after being irradiated. Degree fluorescence) to classify and count cells.
  • Figure 1 is an optical detection device of a blood cell analyzer.
  • Cells pass through the flow chamber one by one under the action of sheath flow.
  • the light emitted by the laser light source is collimated by the lens, it irradiates the cells passing through the flow chamber, and the light irradiates the cells. It will scatter to the surroundings. After collecting the forward scattered light through a collecting lens, it passes through an aperture to limit the angle of the forward scattered light that finally reaches the photodetector.
  • the forward scattered light is limited to a low angle (or Speaking of small angle) forward scattered light-this angle of forward scattered light is generally used to measure cell volume; at the same time, the side light is collected through another collecting lens in the direction perpendicular to the light irradiated to the cell. The side light is reflected and refracted by the dichroic mirror, and the side scattered light in the side light is reflected when passing through the dichroic mirror, and then reaches the corresponding photodetector-the side scattered light is generally used. To measure the surface complexity of the cell, the side fluorescence in the side light is refracted or transmitted, and then passes through a filter to reach the corresponding photodetector. The side fluorescence is generally used to measure the nucleic acid content in the cell.
  • the optical detection device in Figure 1 only has three measurement channels—that is, a low-angle forward scattered light channel, a side scattered light channel, and a side fluorescence channel. Therefore, the cells can only be measured based on the signals obtained by these three measurement channels.
  • Classification and counting which will limit the further scoring and counting of cells to a certain extent, that is, it is impossible to perform more dimensional and more detailed classification and counting, which reduces the classification ability of abnormal cells;
  • the angular forward scattered light channel replaces or adds a high-angle (or large-angle) scattered light channel. You can directly use the photodetector target surface to receive large-angle forward scattered light, but the signal-to-noise ratio obtained in this way is very poor.
  • the size of the device is generally too large. This is due to the selection of the optical path structure.
  • the forward scattered light channel is generally designed as a refractive optical path structure, so this will cause the size of the optical detection device to be too large, especially for the current direction.
  • the scattered light channel is used to collect scattered light in multiple angle ranges (such as low angle and high angle, etc.).
  • the current measurement of RBC and PLT in blood cell analysis generally uses an impedance method or an optical method.
  • the above two methods have some drawbacks.
  • the impedance method for some abnormal samples, such as low-value PLT samples, there is often no obvious boundary between the PLT histogram and the RBC histogram, which makes the algorithm unable to accurately cut PLT and RBC.
  • the histogram can not obtain accurate PLT measurement results.
  • the accuracy of low-value PLT is another important clinical blood routine index, which has become the main disadvantage of impedance measurement.
  • the present invention mainly provides a sample optical detection device, which is described below.
  • the first forward scattered light signal collection component is used to collect forward scattered light in a first angular range, and the forward scattered light is the reflected light generated by the light source irradiating cells passing through the flow cell;
  • the second forward scattered light signal collecting component is used to directly collect forward scattered light in the second angular range, and the forward scattered light is light generated by the light source irradiating cells passing through the flow cell.
  • the sample optical detection device further includes a reflection light blocking component, which is arranged on the optical path of the forward scattered light generated by the cells passing through the flow cell irradiated by the light source, for reflecting forward scattered light in the first angular range To the first forward scattered light signal collection component, and allow the forward scattered light of the second angle range to directly enter the second forward scattered light signal collection component.
  • a reflection light blocking component which is arranged on the optical path of the forward scattered light generated by the cells passing through the flow cell irradiated by the light source, for reflecting forward scattered light in the first angular range To the first forward scattered light signal collection component, and allow the forward scattered light of the second angle range to directly enter the second forward scattered light signal collection component.
  • the reflective light blocking component includes a reflecting mirror for reflecting forward scattered light in a first angular range to the first forward scattered light signal collection component.
  • the reflector is elliptical.
  • the reflective light-shielding component further includes a light-shielding strip for shielding stray light other than the forward scattered light in the second angular range; wherein the reflector is arranged on the light-shielding strip.
  • the reflector is arranged on the light-shielding strip in such a way that its long axis coincides with the light-shielding strip.
  • the sample optical detection device further includes a lens assembly, which is arranged on the optical path of the forward scattered light generated by the cells passing through the flow cell irradiated by the light source, and is used to collect the light in the first and second angular ranges The light is scattered forward and emitted to the reflective light blocking component.
  • a lens assembly which is arranged on the optical path of the forward scattered light generated by the cells passing through the flow cell irradiated by the light source, and is used to collect the light in the first and second angular ranges The light is scattered forward and emitted to the reflective light blocking component.
  • the lens assembly includes one aspheric lens and one spherical lens, or, multiple aspheric lenses, or multiple spherical lenses, or, one aspheric lens and multiple spherical lenses.
  • the effective data aperture of the spherical lens closest to the flow chamber is at least 0.34.
  • the sample optical detection device further includes a straight-blocking diaphragm, which is arranged between the lens assembly and the reflective light-blocking assembly, and is used to block the direct-angle light in the forward scattered light emitted by the lens assembly, And/or, the forward scattered light emitted by the lens assembly is limited to the first angle range and the second angle range.
  • a straight-blocking diaphragm which is arranged between the lens assembly and the reflective light-blocking assembly, and is used to block the direct-angle light in the forward scattered light emitted by the lens assembly, And/or, the forward scattered light emitted by the lens assembly is limited to the first angle range and the second angle range.
  • the first forward scattered light signal collection component includes a first angle limiting diaphragm, a stray light blocking diaphragm, and a photodetector which are sequentially arranged; the first angle limiting diaphragm is used to transmit the reflected light The forward scattered light is limited in the first angular range and converges on the stray light stop, the stray light stop is used to shield the stray light of the forward scattered light in the first angular range, and the photodetector is used To convert the collected forward scattered light in the first angular range into an electrical signal.
  • the second forward scattered light signal collection component includes a stray light blocking diaphragm and a photodetector arranged in sequence, and the stray light blocking diaphragm is used to shield the forward scattered light in the second angular range. Stray light, the photodetector is used to convert the collected forward scattered light in the second angular range into an electrical signal.
  • the sample optical detection device further includes a third forward scattered light signal collection component for collecting forward scattered light in a third angular range, and the forward scattered light is irradiated by the light source through the flow chamber. Light produced by cells and reflected at least once.
  • the third forward scattered light signal collection component includes a mirror, a third angular range aperture diaphragm, and a photodetector which are sequentially arranged; the mirror is used to irradiate the light source through the flow cell The forward scattered light of the third angular range generated by the cell is reflected to the third angular range aperture diaphragm, and the third angular range aperture diaphragm is used to limit the forward scattered light in the third angular range, and the photoelectric The detector is used to convert the collected forward scattered light in the third angular range into an electrical signal.
  • the first angle range is a low angle range
  • the second angle range is a medium angle range
  • the third angle range is a high angle range
  • the first angle range and the second angle range are continuous ranges.
  • the first angle range, the second angle range, and the third angle range are continuous ranges.
  • the first angle range is 0 to 10 degrees or 1 degree to 10 degrees; and/or, the second angle range is 10 degrees to 20 degrees; and/or, the third angle range It is 20 degrees to 70 degrees.
  • the sample optical detection device further includes a light source shaping component for collimating the light beam emitted by the light source and making it converge on the cells passing through the flow chamber.
  • the light source shaping assembly includes a collimating lens and a first cylindrical lens arranged in sequence, the collimating lens is used to collimate the light beam emitted by the light source, and the first cylindrical lens is used for To make the light beam converge at the center of the flow chamber in the direction in which the cells pass.
  • the sample optical detection device further includes a second cylindrical mirror arranged on the exit light path of the first cylindrical mirror, for converging the light beam in a direction perpendicular to the cell passing through, so that the scattered light Are illuminated into the straight stop diaphragm.
  • the sample optical detection device further includes an optical isolator disposed between the collimating lens and the first cylindrical mirror to suppress feedback light.
  • the sample optical detection device further includes:
  • the side scattered light signal collection component is used to collect the side scattered light generated by the light source irradiating cells passing through the flow chamber; and/or,
  • the lateral fluorescence signal collection component is used to collect the lateral fluorescence generated by the light source irradiating cells passing through the flow chamber.
  • the first forward scattered light signal collection component collects the reflected forward scattered light in the first angular range
  • the second forward scattered light signal collection component directly collects or collects the refracted forward scattered light.
  • the transmitted forward scattered light through the design of this optical structure, the longitudinal size of the optical path can be compressed, so that the designed sample optical detection device can be miniaturized.
  • an embodiment of the present invention also provides a sample detection method for detecting red blood cells and platelets in a blood sample, the method including:
  • the red blood cells and platelets in the blood sample to be tested are classified.
  • the diluent can maintain the original shape of the red blood cells and platelets in the blood sample to be tested.
  • the at least two kinds of scattered light signals may include axial light loss, forward scattered light signals, mid-angle scattered light signals, high-angle scattered light signals, side scattered light signals, and backscattered light signals. At least two.
  • the axial light loss, the forward scattered light signal, the middle angle scattered light signal, the high angle scattered light signal, the side scattered light signal, and the back scattered light signal The scattering angles are 0° ⁇ 1°, 1° ⁇ 10°, 10° ⁇ 20°, 20° ⁇ 70°, 70° ⁇ 110° and 110° ⁇ 160°.
  • the at least two scattered light signals may include at least one, especially at least two, of a forward scattered signal, a middle-angle scattering signal, and a high-angle scattering signal.
  • the at least two kinds of scattered light signals may include forward scattering signals and mid-angle scattering signals or may include forward scattering signals and high-angle scattering signals.
  • the light irradiation may be polarized light irradiation
  • the at least two kinds of scattered light signals include axial light loss, forward scattered light signal, intermediate light caused by polarized light irradiation of particles in the sample liquid.
  • the at least two kinds of scattered light signals may include at least one, especially at least two, of a specific polarization state signal of a forward scattering signal, a middle-angle scattering signal, and a high-angle scattering signal.
  • the at least two kinds of scattered light signals may include a specific polarization state signal of a forward scattering signal and a mid-angle scattering signal or may include a specific polarization state signal of a forward scattering signal and a high-angle scattering signal.
  • the step of classifying the red blood cells and platelets in the sample to be tested according to the at least two scattered light signals may include:
  • red blood cells and platelets in the blood sample to be tested are classified.
  • the method may include: obtaining a detection result of predetermined parameters of red blood cells and platelets in the blood sample to be tested based on the at least two scattered light signals.
  • the predetermined parameter may include at least one parameter among red blood cell count, platelet count, average red blood cell volume, average platelet volume, and red blood cell volume distribution width, or a parameter calculated by a combination thereof.
  • the method may further include: outputting classification results of red blood cells and platelets of the blood sample to be tested and/or detection results of predetermined parameters.
  • an embodiment of the present invention also provides a sample analyzer, including:
  • a sampling device having a pipette with a pipette nozzle and a driving device for driving the pipette to quantitatively suck blood samples through the pipette nozzle;
  • the sample liquid preparation device has a reaction tank and a liquid supply part, wherein the reaction tank is used to receive the blood sample drawn by the sampling device, and the liquid supply part supplies the diluent to the reaction tank, so that the sampling device is The sucked blood sample is mixed with the diluent provided by the liquid supply part in the reaction tank to prepare a sample liquid to be tested;
  • the optical detection device has a light source, a flow chamber, and a light collector, wherein the particles in the sample liquid to be tested after the diluent treatment can flow in the flow chamber, and the light emitted by the light source illuminates the flow chamber Particles to generate at least two scattered light signals, and the light collector is used to collect the at least two scattered light signals;
  • a conveying device for conveying the sample solution to be tested that has been processed by the diluent in the reaction tank to the optical detection device;
  • the processor is in communication connection with the sampling device, the sample preparation device, the optical detection device and the conveying device and is used for:
  • the red blood cells and platelets in the blood sample to be tested are classified according to the at least two kinds of scattered light signals.
  • the diluent can maintain the original shape of the red blood cells and the platelets in the blood sample.
  • the at least two kinds of scattered light signals may include at least one of axial light loss, forward scattered light signal, middle angle scattered light signal, high angle scattered light signal, side scattered light signal, and back scattered light signal. Two kinds.
  • the axial light loss, the forward scattered light signal, the middle angle scattered light signal, the high angle scattered light signal, the side scattered light signal, and the back scattered light signal The scattering angles are 0° ⁇ 1°, 1° ⁇ 10°, 10° ⁇ 20°, 20° ⁇ 70°, 70° ⁇ 110° and 110° ⁇ 160°.
  • the at least two scattered light signals may include at least one, especially at least two, of a forward scattered signal, a middle-angle scattering signal, and a high-angle scattering signal.
  • the at least two kinds of scattered light signals may include forward scattering signals and mid-angle scattering signals or may include forward scattering signals and high-angle scattering signals.
  • the light source may be configured as a light source that emits polarized light
  • the at least two scattered light signals include axial light loss and forward scattered light signals generated by the particles in the sample liquid due to polarized light irradiation.
  • the at least two kinds of scattered light signals include at least one, especially at least two, of a specific polarization state signal of a forward scattering signal, a middle-angle scattering signal, and a high-angle scattering signal.
  • the processor may be used to:
  • red blood cells and platelets in the blood sample to be tested are classified.
  • the processor may be configured to obtain a detection result of predetermined parameters of red blood cells and platelets in the blood sample to be tested based on the at least two kinds of scattered light signals.
  • the predetermined parameter may include at least one parameter among red blood cell count, platelet count, average red blood cell volume, average platelet volume, and red blood cell volume distribution width, or a parameter calculated by a combination thereof.
  • the sample analyzer may further include an output device communicatively connected with the processor for outputting the classification results of red blood cells and platelets of the blood sample to be tested and/or the detection results of predetermined parameters.
  • the method for detecting red blood cells and platelets in a blood sample and the sample analyzer according to the embodiments of the present invention can achieve accurate classification of RBC and PLT, especially in a common diluent environment.
  • Figure 1 is a schematic structural diagram of an optical detection device of a blood cell analyzer
  • FIG. 2 is a schematic diagram of the structure of a sample optical detection device in an embodiment
  • FIG. 3 is a schematic diagram for explaining the structure of a first forward scattered light signal collection component and a second forward scattered light signal collection component;
  • FIG. 5 is a schematic structural diagram of a sample optical detection device including a lens assembly in an embodiment
  • Figure 6 is a schematic diagram of a structure for a lens assembly
  • FIG. 7 is a schematic structural diagram of a sample optical detection device including a straight diaphragm in an embodiment
  • FIG. 8 is a schematic structural diagram of a sample optical detection device including a third forward scattered light signal collection component in an embodiment
  • FIG. 9 is a schematic diagram for explaining the structure of a third forward scattered light signal collecting component
  • FIG. 10 is a schematic structural diagram of a sample optical detection device including a side scattered light signal collection component and a side fluorescence signal collection component in an embodiment
  • FIG. 11 is a schematic diagram for explaining the structure of the side scattered light signal collecting component and the side fluorescent signal collecting component
  • Fig. 12 is a schematic structural diagram of a sample optical detection device including a light source shaping component in an embodiment
  • FIG. 13 is a schematic structural diagram of a sample optical detection device according to another embodiment.
  • Figure 14 is a histogram obtained by detecting RBC and PLT using the impedance method
  • Fig. 15A is an RBC histogram obtained by detecting RBC and PLT of normal samples by the impedance method
  • Fig. 15B is a PLT histogram obtained by detecting RBC and PLT of normal samples by impedance method
  • Figure 16 is a PLT histogram obtained by using impedance method to detect RBC and PLT of abnormal samples
  • FIG. 17 is a schematic structural diagram of a sample analyzer provided by an embodiment
  • FIG. 18 is a schematic flowchart of a sample detection method for detecting red blood cells and platelets in a blood sample provided by an embodiment
  • 19 is a schematic diagram of the scattering angles of various scattered light signals in an embodiment
  • FSC forward scattered light signal
  • MAS mid-angle scattered light signal
  • FIG. 21 is a scatter plot of a forward scattered light signal (FSC) and a high-angle scattered light signal (WAS) obtained by a sample detection method provided by an embodiment.
  • FSC forward scattered light signal
  • WAS high-angle scattered light signal
  • connection and “connection” mentioned in this application include direct and indirect connection (connection) unless otherwise specified.
  • an embodiment of the present invention provides a sample optical detection device, the sample optical detection device includes a light source 1, a flow chamber 6, a first forward scattered light signal collection component 20 and a second forward scattered light signal
  • the collection component 30 is described in detail below.
  • the flow chamber 6 is used for the cells in the sample to be tested to pass one by one.
  • the sheath flow technology is used to make the blood cells queue through the flow chamber 6 one by one.
  • the Y-axis direction in the figure is the direction of blood cell movement in the sample. It should be noted that the Y-axis direction in the figure is a direction perpendicular to the paper.
  • the light source 1 is used to illuminate the cells passing through the flow chamber 6.
  • the light source 1 is a laser, such as a helium-neon laser or a semiconductor laser.
  • the first forward scattered light signal collection component 20 and the second forward scattered light signal collection component 30 are respectively used to collect forward scattered light in the first angular range and forward scattered light in the second angular range, This will be explained in detail below.
  • the first forward scattered light signal collection component 20 is used to collect forward scattered light in a first angular range, and the forward scattered light is the reflected light generated by the light source 1 irradiating the cells passing through the flow cell 6.
  • the first forward scattered light signal collection component 20 includes a first angle limiting diaphragm 21, a stray light blocking diaphragm 22, and a photodetector 23 arranged in sequence; the first angle limiting diaphragm is used To limit the reflected forward scattered light in the first angular range and converge on the stray light stop 22, the stray light stop 22 is used to shield the stray light of the forward scattered light in the first angular range, and photodetection The device 23 is used to convert the collected forward scattered light in the first angular range into an electrical signal.
  • the first angle range is a low angle range, for example, the first angle range is 0 to 10 degrees or 1 degree to 10 degrees.
  • the second forward scattered light signal collection component 30 is used to collect forward scattered light in a second angular range, and the forward scattered light is light generated by the light source 1 irradiating cells passing through the flow cell 6.
  • the second forward scattered light signal collection component 30 includes a stray light blocking diaphragm 31 and a photodetector 32 which are sequentially arranged, and the stray light blocking diaphragm 31 is used to shield the forward scattered light in the second angular range. Stray light, the photodetector 32 is used to convert the collected forward scattered light in the second angular range into an electrical signal.
  • the second angle range is a middle angle range, for example, the second angle range is 10 degrees to 20 degrees.
  • the first angle range and the second angle range are continuous ranges.
  • the first forward scattered light signal collection component 20 collects the reflected forward scattered light in the first angular range, and the second forward scattered light signal collection component 30 directly collects or collects the refracted or transmitted forward scattered light,
  • the longitudinal dimension of the optical path (the Z axis direction in the figure) can be compressed, so that the designed sample optical detection device can be miniaturized.
  • a reflective light blocking component is used to spatially separate the forward scattered light in the first angular range and the second angular range, and one is reflected to the first forward scattered light signal collection component 20 , One is allowed to pass through to the second forward scattered light signal collection component 30, which will be described in detail below.
  • the sample optical detection device in an embodiment further includes a reflective light blocking component 10, which is disposed on the optical path of the forward scattered light generated by the light source 1 irradiating the cells passing through the flow chamber 6, and is used to adjust the first angle range
  • the forward scattered light is reflected to the first forward scattered light signal collection component 20, and the forward scattered light in the second angular range is allowed to directly enter the second forward scattered light signal collection component 30.
  • the reflective light blocking assembly 10 includes a mirror 11 for reflecting forward scattered light in a first angular range to the first A forward scattered light signal collection component 20.
  • the reflector 11 may be elliptical, that is, the reflector 11 is an elliptical aperture reflector.
  • the placement angle of the reflector 11 can be 45 degrees.
  • the mirror surface is parallel to the Y axis in the figure.
  • the placement angle of the reflector 11 can also be adjusted according to the optical path spatial layout, and is not limited to only 45 degrees. Please refer to FIG. 4( b ).
  • the reflective light blocking component 10 includes a light blocking strip 12 for shielding stray light except for the forward scattered light in the second angle range.
  • the reflector 11 is arranged on the light-shielding strip 12.
  • the reflector 11 when the reflector 11 is elliptical, the reflector 11 is arranged on the light-shielding strip 12 in such a way that its long axis coincides with the light-shielding strip 12, as shown in Figure 4 (c ) As shown.
  • the shading strip 12 here acts as a support for the reflector 11, and the other is to shield stray light except for the forward scattered light in the second angle range, which can effectively ensure the signal quality of the forward scattered light in the second angle range. .
  • the sample optical detection device may further include a lens assembly 13, which is arranged in the light source 1 to illuminate The forward scattered light generated by the cells in the flow chamber 6 is used to collect the forward scattered light in the first angular range and the second angular range on the optical path, and emit the forward scattered light to the reflective light blocking assembly 10.
  • the lens assembly 13 includes one aspheric lens and one spherical lens, or, multiple aspheric lenses, or multiple spherical lenses, or, one aspheric lens and multiple spherical lenses. Please refer to FIG.
  • the lens assembly 13 can be composed of two spherical lenses 14 and 15 in a specific embodiment.
  • the effective data aperture of the spherical lens closest to the flow chamber 6 is at least 0.34.
  • the introduction of the lens assembly 13 can ensure that the system aberrations are effectively corrected, and that the signal spot can be effectively collected by the two measurement channels of the first forward scattered light signal collection component 20 and the second forward scattered light signal collection component 30 .
  • the combination of the lens assembly 13 and the reflective light blocking assembly 10 can effectively ensure the signal quality of the forward scattered light in the first angular range.
  • the sample optical detection device may further include a straight stop 16 arranged at the lens assembly 13 and Reflective light blocking components 10 between.
  • the straight-blocking diaphragm 16 has many functions.
  • the straight-blocking diaphragm 16 can block the direct angle of 0 degree light in the forward scattered light emitted by the lens assembly 13-this can prevent signal saturation, and/or can prevent the lens
  • the forward scattered light emitted by the component 13 is limited to the first angle range and the second angle range.
  • the sample optical detection device can collect forward scattered light in the first angle range and the second angle range.
  • the sample optical detection device can also collect forward scattered light in the third angle range.
  • the third angle range is a high angle range, for example, the third angle range is 20 degrees to 70 degrees.
  • the first angle range, the second angle range, and the third angle range are continuous ranges. The following specifically describes how the sample optical detection device collects forward scattered light in the third angular range.
  • the optical path space behind the flow chamber 6 in the sample optical detection device is relatively tight-for example, it is provided with a first forward scattered light signal collection component 20 and a second forward scattered light signal collection component 30, and even a lens Components 13 and so on. Therefore, if a photodetector is used to directly collect forward scattered light, it cannot guarantee that the forward scattered light in the third angle range (for example, high angle range) can be completely collected. Therefore, the inventor considers the optical path structure for refraction To convert the forward scattered light in the third angle range (for example, the high angle range), so that the forward scattered light in the third angle range (for example, the high angle range) can be effectively collected.
  • the sample optical detection device further includes a third forward scattered light signal collection component 40 for collecting forward scattered light in a third angular range, and the forward scattered light is irradiated by the light source 1 The light generated by the cells of the flow chamber 6 and reflected at least once.
  • the third forward scattered light signal collection component 40 includes a mirror 41, a third angular range aperture stop 42 and a photodetector 43 arranged in sequence; the mirror 41 is used to connect the light source 1 The forward scattered light of the third angular range generated by the cells irradiated through the flow cell 6 is reflected to the third angular range aperture stop 42, and the third angular range aperture stop 42 is used to limit the forward scattered light to the third angular range The photodetector 43 is used to convert the collected forward scattered light in the third angular range into an electrical signal.
  • the sample optical detection device shown in FIG. 9 is only an example drawn to illustrate the third forward scattered light signal collection component 40, which does not represent the sample optical detection device including the third forward scattered light signal collection component 40 The detection device can only have the structure shown in FIG. 9.
  • the sample optical detection device of the present invention can also realize the detection of side light, such as side scattered light and/or side fluorescence. Collection is described in detail below with reference to Figure 10 and Figure 11.
  • the sample optical detection device of an embodiment further includes a side scattered light signal collection assembly 50 for collecting side scattered light generated by the light source 1 irradiating the cells passing through the flow chamber 6.
  • the side scattered light signal collection component 50 includes a side small aperture diaphragm 51 and a photodetector 52 arranged in sequence.
  • the side small aperture diaphragm 51 is used to process lateral scattered light, and the photodetector 52 receives The lateral dispersion light processed by the lateral aperture diaphragm 51 is converted into an electrical signal.
  • the sample optical 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 cells passing through the flow chamber 6.
  • the lateral fluorescence signal collection component 60 includes a fluorescent aperture diaphragm 61, a filter 62, and a photodetector 63 arranged in sequence.
  • the fluorescent aperture diaphragm 61 is used to process lateral fluorescence, and the processed lateral The fluorescence passes through the filter 62 and reaches the photodetector 63, which is used to convert the lateral fluorescence into an electrical signal.
  • a collecting lens 7 can be set on the path of the side light of the flow chamber 6 to collect the side light (including side scattered light and side fluorescence), and the dichroic mirror 8
  • the side scattered light is reflected to the side scattered light signal collection component 50—for example, the dichroic mirror 8 reflects and focuses the side scattered light on the side aperture diaphragm 51, and then enters the photodetector 52; the side fluorescence is transparent Pass the dichroic mirror 8 and enter the lateral fluorescence signal collection component 60-for example, the lateral fluorescence is focused on the fluorescence aperture diaphragm 61 through the dichroic mirror 8 and then reaches the photodetector 63 after passing through the filter 62 .
  • the side scattered light and the side fluorescence in an embodiment may be 70 degrees to 110 degrees of light.
  • FIGS. 10 and 11 are an example in which the sample optical detection device includes both a side scattered light signal collection component 50 and a side fluorescence signal collection component 60.
  • the sample optical detection device may only It includes one of a side scattered light signal collecting component 50 and a side fluorescent signal collecting component 60.
  • the sample optical detection device may also include a light source shaping component 9 for collimating the light beam emitted by the light source 1 and converging it to the flow Cells of chamber 6. Referring to FIG.
  • 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 arranged in sequence; the collimating lens 2 is used to align the light source 1 The emitted light beam is collimated, and the optical isolator 3 is used to suppress the feedback light and prevent the reflected light from the subsequent optical devices from entering the light source 1 and affecting the light source 1.
  • the first cylindrical mirror 4 is used to make the light beam pass through the cell Direction converges at the center of the flow chamber, for example, the first cylindrical mirror 4 makes the light beam converge at the center of the flow chamber in the Y-axis direction in the figure; the second cylindrical mirror 5 is used to direct the light beam perpendicular to The cells converge in the passing direction.
  • the second cylindrical mirror 5 is used to converge the light beam in the X-axis direction in the figure, so that all scattered light is irradiated into the straight stop 16.
  • the components in the light source shaping assembly 9 are not necessary in some cases, such as the optical isolator 3 and the second cylindrical mirror 5, etc.
  • Figure 13 above is a schematic diagram of a sample optical detection device with five measurement channels. Take Figure 13 as an example below to explain the working process and principles of these five measurement channels.
  • the flow direction of the sample cells (that is, the direction through the flow chamber 6) is the Y-axis direction, that is, the direction perpendicular to the paper.
  • the light emitted from the light source 1 passes through the collimator lens 2 to become an equal beam, passes through the optical isolator 3, and then passes through the first cylindrical mirror 4 to make the light beam converge in the direction in which the sample cells flow, that is, in the Y-axis direction.
  • the center of the flow chamber to illuminate the cells passing through the flow chamber 6.
  • the main function of the optical isolator 3 is to suppress the feedback light-the feedback light mainly comes from the reflection of the optical components behind the optical isolator 3-enters the light source 1, preventing the output power of the light source 1 from fluctuating, and ensuring the optical baseline stability.
  • the second cylindrical lens 5, spherical lenses 14 and 15 will make the beam converge in the X-axis direction.
  • the second cylindrical lens 5, spherical lenses 14 and 15 to make the size of the beam in the X-axis direction Compressed to be smaller than the horizontal size of the straight bar of the straight stop 16 to ensure that the direct light can be effectively blocked by the straight stop 16 to prevent signal saturation;
  • the straight bar of the straight stop 16 is A straight bar along the Y-axis direction, that is, the axial direction of the straight bar is the Y-axis direction, and the lateral direction is the X-axis direction.
  • the light beam from the light source 1 will finally irradiate the cells passing through the flow chamber 6, producing forward scattered light in the first angular range (the low-angle forward scattered light may be used as an example below. ), the forward scattered light in the second angle range (the forward scattered light at the middle angle may be taken as an example below), the forward scattered light in the third angle range (the forward scattered light at a high angle may be taken as an example below), Side scattered light and side fluorescence:
  • the low-angle forward scattered light and the medium-angle forward scattered light are collected by spherical lenses 14 and 15, and then the low-angle forward scattered light is reflected by the reflective light blocking component 10 (such as its elliptical aperture mirror) to the first
  • the angle limiting aperture 21 (for example, the low angle limiting aperture) converges at the stray light stop 22, and then enters the photodetector 23; while the forward scattered light of the medium angle is reflected by the light blocking component 10 (for example, it is blocked Strip 12, the shading strip 12 is arranged along the plane direction composed of the X axis and the Z axis, and the effective shielding direction is the X axis direction), which is effectively shielded from the stray light of the forward scattered light at the middle angle, and then converges on the shield
  • the stray light diaphragm 31 then enters the photodetector 32;
  • the high-angle forward scattered light is reflected by the reflector 41 to the third angular range aperture stop 42 (for example, the high-angle aperture stop), and then enters the photodetector 43;
  • the side scattered light and the side fluorescence are collected by the collecting lens 7 at the same time, and are spatially separated when passing through the dichroic mirror 8, wherein the side scattered light is reflected and focused on the side small hole through the front surface of the dichroic mirror 8.
  • the diaphragm 51 then enters the photodetector 52; the lateral fluorescence passes through the dichroic mirror 8 and then focuses on the fluorescent aperture diaphragm 61, and then passes through the filter 62 to reach the photodetector 63.
  • the present invention uses lens components to collect low- and mid-angle forward scattered light in forward scattered light, and uses reflective light blocking components to spatially separate low-angle forward scattered light and mid-angle forward scattered light, which not only ensures low-angle
  • the forward scattered light is collected, which can replace the traditional classification of eosinophils through the side scattered light signal; the present invention adds a medium-angle forward scattered light measurement channel and High-angle forward scattered light measurement channels, through these three measurement channels, some cells can be classified and counted.
  • platelets can be removed from red blood cells without using fluorescent or spheronizing reagents.
  • Classification the present invention uses a collection lens set on the side light path of the flow chamber to depolarize and collect the fluorescent signal of the side light, and separate the depolarized side scattered light and the fluorescent signal through the wavelength selection of the dichroic mirror .
  • the present invention realizes the optical path structure for receiving scattered light from 0 degree or 1 degree to 110 degrees, and there is no lost angle in the middle.
  • impedance method or fluorescence optical method is generally used in blood analyzers to measure RBC and PLT in blood samples.
  • the impedance method is based on the Coulter principle.
  • the diluted blood sample is passed through a small hole, and a constant current source is applied to both sides of the small hole.
  • Each cell passing through the small hole causes a change in the electrical impedance of the liquid in the small hole, thereby generating Electric pulse.
  • the histogram is one-dimensional information, that is, only the volume information of the cell, as shown in Figure 14.
  • the left side of the dashed line is the PLT histogram
  • the right side of the dashed line is the RBC histogram. Since the diameter of RBC particles is about 3 times that of PLT, and the number of RBC particles is about 30 times that of PLT, the PLT histogram is on the left side of the RBC histogram, and the area enclosed by the horizontal axis is much smaller than that of RBC.
  • the dotted line in FIG. 14 is the dividing line of the histogram of RBC and PLT.
  • FIG. 15A is the impedance method.
  • the fluorescence optical method can overcome this defect.
  • the fluorescence method is based on flow cytometry. The diluted and stained samples are squeezed by the sheath flow through the optical detection zone in sequence. Each cell is irradiated by the excitation light source, and the forward scatter signal (representing the cell volume) and the fluorescence signal (representing the nucleic acid content in the cell) are obtained in the optical system to generate a two-dimensional scatter plot to divide the RBC and PLT And calculation.
  • the two-dimensional scatter plot of the optical method has one more dimension of information, so that PLT and RBC can be accurately divided on the two-dimensional scatter plot.
  • fluorescent dyes are needed to stain the cells, and a special diluent is needed to spheronize the cells, so the cost is high, which is not conducive to the promotion of optical methods in clinical practice.
  • the present invention proposes a sample detection method and sample analyzer that only use scattered light information to detect red blood cells and platelets in blood samples.
  • the sample detection method and sample analyzer can be implemented in a common diluent environment
  • the accurate classification of RBC and PLT in particular, can realize the accurate classification of RBC and PLT of abnormal samples in the environment of ordinary diluent.
  • the sample analyzer 100 at least includes a sampling device (not shown), a sample liquid preparation device 110, an optical detection device 120, a conveying device 130 and a processor 140.
  • the sampling device has a pipette with a pipette nozzle and has a driving device for driving the pipette to quantitatively suck blood samples through the pipette nozzle. Further, the sampling device is driven by its driving device after the blood sample is drawn and moved to the reaction cell 111 of the sample liquid preparation device 110, and the drawn blood sample is injected into the reaction cell 111.
  • the sample liquid preparation device 110 has at least one reaction cell 111 and has a liquid supply part (not shown), wherein the reaction cell 111 is used to receive a blood sample drawn by the sampling device, and the liquid supply part dilutes the liquid It is provided to the reaction tank 111, so that the blood sample drawn by the sampling device and the diluent provided by the liquid supply part react in the reaction tank to prepare a blood sample to be tested.
  • the liquid supply part can be used to inject an appropriate diluent into the reaction tank to process particles in the blood sample to prepare a blood sample to be tested for subsequent testing.
  • the diluent is a common diluent necessary for a blood cell analyzer to maintain the original form of the red blood cells and the platelets in the blood sample, and there is no need to use special diluents that make RBC and PLT spheroidize.
  • the diluent may include ingredients such as sodium chloride, phosphate buffer, preservative, etc.
  • the diluent is not limited to one, and can be selected according to needs, and will not be repeated here.
  • the optical detection device 120 has a light source 121, a flow chamber 122 and light collectors 123 and 124.
  • the light source 121 can emit natural light or light of a specific wavelength band, and is not limited to one. Alternatively, the light source 121 can also be a polarized light source to emit polarized light in a specific polarization state.
  • the flow chamber 122 has an orifice 1221, and the particles of the sample liquid to be tested after being processed by the diluent in the sample liquid preparation device 110 can flow in the flow chamber 122 and pass through the orifice 1221 one by one. The light emitted by the light source 121 irradiates the particles in the flow chamber 122 to generate optical signal information.
  • the light collectors 123 and 124 are used to collect the optical signal information.
  • the optical signal information may include at least two of axial light loss, forward scattered light signal, mid-angle scattered light signal, high-angle scattered light signal, side scattered light signal, and back scattered light signal.
  • the optical signal information includes the axial light loss, the forward scattered light signal, the middle angle scattered light signal, the high angle scattered light signal, the side scattered light signal, and the At least two of the signals of the specific polarization state of the backscattered light signal.
  • the optical detection device 120 includes at least an axial light loss concentrator, a forward scattered light signal concentrator, a middle angle scattered light signal concentrator, a high angle scattered light signal concentrator, and a side scattered light signal concentrator. And at least two light collectors in the backscattered light signal light collector.
  • the light collector is configured as a photodetector, such as a photodiode or a photomultiplier tube.
  • a photodetector such as a photodiode or a photomultiplier tube.
  • the forward scattered light emitted by the blood cells flowing in the flow chamber 122 is received by the photodiode (forward scattered light collector) 123 through the condenser 126 and the pinhole 127, and the side scattered light It is received by the photomultiplier tube (side scattered light concentrator) 124 through the condenser lens 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 respectively sent to the processor 140 after being amplified by the amplifier 150 and analog signal processing such as waveform processing.
  • the axial light loss, the forward scattered light signal, the middle angle scattered light signal, the high angle scattered light signal, the side scattered light signal, and the back scattered light signal The scattering angles are 0° ⁇ 1°, 1° ⁇ 10°, 10° ⁇ 20°, 20° ⁇ 70°, 70° ⁇ 110° and 110° ⁇ 160°.
  • the axial light loss concentrator, the forward scattered light signal concentrator, the middle angle scattered light signal concentrator, the high angle scattered light signal concentrator, and the side scattered light signal concentrator are respectively configured to receive the aforementioned scattered light. Angle of scattered light signal.
  • the optical detection device 120 includes at least one, especially at least two, of a forward scattered light signal concentrator, a middle-angle scattered light signal concentrator, and a high-angle scattered light signal concentrator, so as to receive At least one of the forward scatter signal, the middle-angle scatter signal, and the high-angle scatter signal, especially at least two, or at least one of the forward scatter signal, the middle-angle scatter signal, and the specific polarization state signal of the high-angle scatter signal, especially It is at least two kinds of scattered light signals to improve the accuracy of the detection results of the red blood cells and platelets.
  • the optical detection device 120 includes a forward scattered light signal collector and a middle-angle scattered light signal collector or a forward scattered light signal collector and a high-angle scattered light signal collector.
  • the conveying device 130 is used to convey the blood sample to be tested that has been processed by the diluent in the reaction tank 111 to the optical detection device 120.
  • the processor 140 is connected to the sampling device, the sample preparation device 110, the optical detection device 120, and the transport device communication 130, and is configured to acquire the optical signal information from the optical detection device 120 and process the optical signal information , In order to obtain the results of particle classification and counting 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.
  • the processor 140 is configured to implement the sample detection method according to the present invention, which will be described in detail below.
  • the aforementioned processor 140 may be an application-specific integrated circuit (Application Specific Integrated Circuit, ASIC), Digital Signal Processor (DSP), Digital Signal Processing Device (Digital Signal Processing Device, DSPD), Programmable Logic Device (ProgRAMmable Logic Device, PLD), Field Programmable Gate Array (Field Programmable Gate Array) At least one of ProgRAMmable Gate Array (FPGA), Central Processing Unit (CPU), controller, microcontroller, and microprocessor. It can be understood that, for different devices, the electronic devices used to implement the above-mentioned processor functions may also be other, which is not specifically limited in the embodiment of the present invention.
  • ASIC Application Specific Integrated Circuit
  • DSP Digital Signal Processor
  • DSPD Digital Signal Processing Device
  • PLD Programmable Logic Device
  • Field Programmable Gate Array Field Programmable Gate Array
  • FPGA Field Programmable Gate Array
  • CPU Central Processing Unit
  • controller microcontroller, and microprocessor
  • Figure 18 is a sample detection method for detecting red blood cells and platelets in a blood sample provided by an embodiment.
  • a sample detection method 200 for detecting red blood cells and platelets in a blood sample includes the following steps:
  • step S210 a sample solution to be tested containing a blood sample to be tested and a diluent is prepared.
  • the sample liquid to be tested is prepared in the sample preparation device 110 of the sample analyzer 100.
  • the sample liquid preparation device 110 has at least one reaction cell 111 and a liquid supply part (not shown).
  • the sampling device is driven by its driving device after the blood sample is drawn and moved to the reaction cell 111 of the sample liquid preparation device 110, and then the liquid supply part provides the diluent to the reaction cell 111, so that the sampling device The blood sample sucked by the device reacts with the diluent provided by the liquid supply part in the reaction tank to prepare a blood sample to be tested.
  • the diluent provides the appropriate pH, conductivity and osmotic pressure for the blood sample to be tested to ensure the integrity of the cells without hemolysis; in addition, the diluent is also used to clean the remaining substances in the last test sample , To ensure the cleanliness of the sampling needle, pipeline and flow chamber of the sample analyzer to prevent cross contamination; on the other hand, the diluent should be filled in the middle of the pipeline during the temporary stop of the sample analyzer to prevent foreign matter such as external dust from entering the instrument, causing malfunction.
  • the diluent is a common diluent necessary for a blood cell analyzer to maintain the original shape of the red blood cells and the platelets in the blood sample, rather than a special diluent that makes RBC and PLT spherical.
  • the diluent may include ingredients such as sodium chloride, phosphate buffer, and preservative.
  • the diluent is not limited to one, and can be selected according to needs. Repeat it again.
  • Step S220 irradiate the sample liquid to be tested with light in the optical detection area.
  • the transport device 130 of the sample analyzer 100 transports the blood sample to be tested after the diluent treatment in the reaction cell 111 to the flow cell 122 of the optical detection device 120,
  • the particles of the sample solution processed by the diluent can flow in the flow chamber 122 and pass through the aperture 1221 one by one.
  • the light emitted by the light source 121 irradiates the particles in the flow chamber 122 to generate optical signal information.
  • Step S230 Collect at least two kinds of scattered light signals generated by light irradiation of particles in the sample liquid to be tested, that is, scattered light signals with specific scattering angles.
  • the optical detection device 120 transmits the scattered light signal output by the optical detection device 120 to the processor 140, so that the processor 140 can process the scattered light signal.
  • the scattered light signal may include axial light loss, forward scattered light signal, mid-angle scattered light signal, high-angle scattered light signal, side scattered light signal, and backscattered light signal. At least two.
  • the scattered light signal includes at least one, especially at least two, of a forward scattering signal, a middle-angle scattering signal, and a high-angle scattering signal. More preferably, the at least two kinds of scattered light signals include a forward scattering signal and a mid-angle scattering signal or include a forward scattering signal and a high-angle scattering signal.
  • the scattering angle refers to the center of the overlapping area between the sample flow and the excitation beam in the flow chamber as the vertex of the angle, and the first side of the angle with the propagation direction of the excitation beam as The propagation direction of the scattered light emitted by the particles at the apex is the second side of the angle, the angle formed by the apex and the two sides. If there is no explanation to the contrary, the scattering angle shall refer to this explanation.
  • the present invention defines the scattering angle of the scattered light signal as follows, as shown in FIG. 19, wherein: the scattering angle of the axial light loss is: 0° ⁇ 1°; the forward scattered light signal The scattering angle of the medium-angle scattered light signal is: 1° ⁇ 10°; the scattering angle of the medium-angle scattered light signal is: 10°-20°; the scattering angle of the high-angle scattered light signal is: 20° ⁇ 70°; the lateral direction The scattering angle of the scattered light signal is 70° ⁇ 110°; and the scattering angle of the backscattered light signal is 110° ⁇ 160°.
  • any two or all of the forward scatter signal, the mid-angle scatter signal, and the high-angle scatter signal are collected for subsequent classification of red blood cells and platelets in the blood sample, and the above three are selected.
  • At least two of the scattered light signals can more effectively and accurately classify red blood cells and platelets.
  • using at least two of the forward scatter signal, the mid-angle scatter signal, and the high-angle scatter signal to obtain a scatter diagram of red blood cells and platelets there are clearer red blood cell and platelet boundaries, and the result of classification More precise.
  • the light irradiation may be natural light or light of a specific wavelength band, and polarized light irradiation of a specific polarization state may also be selected.
  • the scattered light signal is the axial light loss, forward scattered light signal, and mid-angle scattered light signal generated by the particles in the sample liquid due to the polarized light irradiation. , High-angle scattered light signals, side scattered light signals, and backscattered light signals with specific polarization states.
  • any two or all of the specific polarization state signals of the forward scatter signal, the mid-angle scatter signal, and the high-angle scatter signal are collected for subsequent analysis of the red blood cells and the red blood cells in the blood sample. Platelets are classified to obtain a more effective and accurate classification of red blood cells and platelets.
  • step S240 the red blood cells and platelets in the blood sample to be tested are classified according to the at least two kinds of scattered light signals.
  • the processor 140 receives and processes the scattered light signal from the optical detection device 120 to obtain a classification result of red blood cells and platelets in the blood sample to be tested.
  • the step of classifying the red blood cells and the platelets may include: generating a two-dimensional or three-dimensional scatter plot of particles in the blood sample according to the at least two kinds of scattered light signals; Based on the two-dimensional or three-dimensional scatter plot, red blood cells and platelets in the blood sample are classified.
  • Fig. 20 is a scatter plot of forward scatter (FSC) light signal and mid-angle scatter (MAS) light signal.
  • Figure 21 is a scatter plot of forward scatter (FSC) light signal and high angle scatter (WAS) light signal.
  • the detection results of predetermined parameters of the red blood cells and the platelets can also be obtained based on the at least two kinds of scattered light signals.
  • the predetermined parameters may include at least one parameter among red blood cell count, platelet count, average red blood cell volume MCV, average platelet volume MPV, and red blood cell volume distribution width RDW, and other parameters obtained by combining the foregoing parameters.
  • the method 200 may further include outputting the classification and counting results of red blood cells and platelets of the blood sample to be tested and/or the detection results of the predetermined parameters.
  • 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 the accurate classification of RBC and PLT, especially in the environment of ordinary diluent.
  • the red blood cells and platelets in the blood sample maintain the original shape, for example, the red blood cells maintain the shape of a double-sided concave disc, with thick edges and thinner middles, so it is difficult to detect and distinguish.
  • this defect can be compensated.
  • only scattered light is used to distinguish red blood cells and platelets.

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Abstract

一种样本光学检测装置，包括：流动室，用于供待测样本中的细胞逐个通过；光源，用于照射通过所述流动室的细胞；第一前向散射光信号收集组件，用于收集第一角度范围的前向散射光，该前向散射光为所述光源照射通过流动室的细胞产生的且经反射后的光；第二前向散射光信号收集组件，用于直接收集第二角度范围的前向散射光，该前向散射光为所述光源照射通过流动室的细胞产生的光。本发明还涉及一种样本检测方法和一种样本分析仪。

Description

样本光学检测装置、样本检测方法及样本分析仪 技术领域
本发明涉及一种样本光学检测装置、一种样本检测方法及一种样本分析仪。
 背景技术
血细胞分析仪大多采用激光散射原理进行测量，原理为：将激光照射在细胞上，通过收集细胞被照射后产生的前向散射光、侧向散射光（90度散射光）和侧向荧光（90度荧光），来对细胞进行分类和计数等。
图1为一种血细胞分析仪的光学检测装置，细胞在鞘流的作用下逐个通过流动室，当激光光源发出的光被透镜准直后向通过流动室的细胞照射，照射到细胞上的光会向四周产生散射，通过一收集透镜来收集前向散射光后，再经过一个光阑来限定最终到达光电探测器的前向散射光的角度，例如将前向散射光限定为低角度（或者说小角度）的前向散射光——这种角度的前向散射光一般用于测量细胞体积；同时，在与照射到细胞的光线垂直的方向通过另一收集透镜来收集侧向光，收集的侧向光再通过二向色镜发生反射和折射，其中侧向光中的侧向散射光在经过二向色镜时发生反射，然后到达相应的光电探测器——侧向散射光一般用于测量细胞的表面复杂程度，侧向光中的侧身荧光则经过折射或者说透射后再经过一滤光片也到达相应的光电探测器——侧向荧光一般用于测量细胞内核酸含量。
 技术问题
图1中的光学检测装置仅有三路测量通道——即低角度前向散射光通道、侧向散射光通道和侧向荧光通道，因此只能基于这三路测量通道获取的信号来对细胞进行分类和计数，这在一定程度会限制对细胞的进一步分数和计数，即无法做到进行更多维度和更加细致的分类和计数，降低了异常细胞的分类能力；技术人员如果将图1中低角度前向散射光通道替换或增加高角度（或者说大角度）散射光通道，可以直接使用光电探测器靶面来接收大角度前向散射光，但这样接收得到的信号信噪比非常差，因此为了保证信号质量，技术人员通常会采用复杂的多个透镜组合来收集大角度前向散射光再出射给对应的光电探测器，这种做法则会极大增加装置的成本；另外，光学检测装置的尺寸一般偏大，这是由于其光路结构所选成的，例如前向散射光通道一般被设计为折射式的光路结构，因此这会造成光学检测装置的尺寸偏大，尤其是当前向散射光通道用于收集多个角度范围（例如低角度和高角度等）的散射光时。
此外，目前在血细胞分析中进行RBC和PLT的测量时，一般采用阻抗法或光学法。但是上述两种方法均存在一些弊端，例如在阻抗法中对于某些异常样本、如低值PLT样本，其PLT直方图与RBC直方图交界处往往没有明显界限，导致算法无法准确切割PLT与RBC直方图，进而不能获取准确的PLT测量结果。而低值PLT的准确性又是临床重点关注的血常规指标，这成为阻抗法测量的主要缺点。另外，在光学法中，需要荧光染料对细胞染色，而且还需要使用特殊的稀释液对细胞进行球形化处理，所以使用成本较高，不利于光学法在临床的推广。
 技术解决方案
本发明主要提供一种样本光学检测装置，下面说明。
一实施例的样本光学检测装置，包括：
流动室，用于供待测样本中的细胞逐个通过；
光源，用于照射通过所述流动室的细胞；
第一前向散射光信号收集组件，用于收集第一角度范围的前向散射光，该前向散射光为所述光源照射通过流动室的细胞产生的且经反射后的光；
第二前向散射光信号收集组件，用于直接收集第二角度范围的前向散射光，该前向散射光为所述光源照射通过流动室的细胞产生的光。
一实施例中，样本光学检测装置还包括反射挡光组件，设置于所述光源照射通过流动室的细胞产生的前向散射光的光路上，用于将第一角度范围的前向散射光反射到所述第一前向散射光信号收集组件，以及允许第二角度范围的前向散射光直接入射到所述第二前向散射光信号收集组件。
一实施例中，所述反射挡光组件包括反射镜，用于将第一角度范围的前向散射光反射到所述第一前向散射光信号收集组件。
一实施例中，所述反射镜为椭圆形。
一实施例中，所述反射挡光组件还包括遮光条，用于屏蔽除第二角度范围的前向散射光外的杂光；其中所述反射镜设置于该遮光条上。
一实施例中，所述反射镜以长轴与所述遮光条重合的方式设置于所述遮光条上。
一实施例中，所述样本光学检测装置还包括透镜组件，设置于所述光源照射通过流动室的细胞产生的前向散射光的光路上，用于收集第一角度范围和第二角度范围的前向散射光，并出射给所述反射挡光组件。
一实施例中，所述透镜组件包括一个非球面透镜和一个球面透镜，或者，多个非球面透镜，或者，多个球面透镜，或者，一个非球面透镜和多个球面透镜。
一实施例中，当所述透组件包括球面透镜时，最靠近流动室的球面透镜的有效数据孔径至少为0.34。
一实施例中，所述样本光学检测装置还包括挡直光阑，设置于所述透镜组件和反射挡光组件之间，用于遮挡透镜组件出射的前向散射光中的直射角度的光，和/或，将透镜组件出射的前向散射光限定在所述第一角度范围和第二角度范围。
一实施例中，所述第一前向散射光信号收集组件包括依次设置的第一角度限定光阑、挡杂光光阑和光电探测器；所述第一角度限定光阑用于将经反射的前向散射光限定在第一角度范围并会聚于所述挡杂光光阑，所述挡杂光光阑用于屏蔽第一角度范围的前向散射光的杂光，所述光电探测器用于将收集的第一角度范围的前向散射光转换成电信号。
一实施例中，所述第二前向散射光信号收集组件包括依次设置的挡杂光光阑和光电探测器，所述挡杂光光阑用于屏蔽第二角度范围的前向散射光的杂光，所述光电探测器用于将收集的第二角度范围的前向散射光转换成电信号。
一实施例中，所述样本光学检测装置还包括第三前向散射光信号收集组件，用于收集第三角度范围的前向散射光，该前向散射光为所述光源照射通过流动室的细胞产生的且经至少一次反射后的光。
一实施例中，所述第三前向散射光信号收集组件包括依次设置的反射镜、第三角度范围孔径光阑和光电探测器；所述反射镜用于将所述光源照射通过流动室的细胞经产生的第三角度范围的前向散射光反射至所述第三角度范围孔径光阑，所述第三角度范围孔径光阑用于前向散射光限定在第三角度范围，所述光电探测器用于将收集的第三角度范围的前向散射光转换成电信号。
一实施例中，所述第一角度范围为低角度范围，和/或，所述第二角度范围为中角度范围，和/或，所述第三角度范围为高角度范围。
一实施例中，所述第一角度范围和第二角度范围为连续范围。
一实施例中，所述第一角度范围、第二角度范围和第三角度范围为连续范围。
一实施例中，所述第一角度范围为0到10度或1度到10度；和/或，所述第二角度范围为10度到20度；和/或，所述第三角度范围为20度到70度。
一实施例中，所述样本光学检测装置还包括光源整形组件，用于对所述光源发出的光束进行准直并使之会聚于通过所述流动室的细胞。
一实施例中，所述光源整形组件包括依次设置的准直透镜和第一柱面镜，所述准直透镜用于对所述光源发出的光束进行准直，所述第一柱面镜用于使光束在细胞通过的方向上会聚于所述流动室的中心。
一实施例中，所述样本光学检测装置还包括设置于所述第一柱面镜的出射光路上的第二柱面镜，用于将光束在垂直于细胞通过的方向上会聚，使得散射光都被照射到所述挡直光阑内。
一实施例中，所述样本光学检测装置还包括设置于所述准直透镜和第一柱面镜之间的光隔离器，用于抑制反馈光。
一实施例中，所述样本光学检测装置还包括：
侧向散射光信号收集组件，用于收集所述光源照射通过流动室的细胞产生的侧向散射光；和/或,
侧向荧光信号收集组件，用于收集所述光源照射通过流动室的细胞产生的侧向荧光。
依据上述实施例的样本光学检测装置，第一前向散射光信号收集组件收集经反射后的第一角度范围的前向散射光，第二前向散射光信号收集组件则直接收集或收集经折射或透射的前向散射光，通过这种光学结构的设计，可以压缩光路的纵向尺寸，使得设计出来的样本光学检测装置小型化。
此外，本发明实施例还提供了一种检测血液样本中的红细胞和血小板的样本检测方法，所述方法包括：
制备含待测血液样本和稀释液的待测样本液；
在光学检测区对所述待测样本液进行光照射；
收集所述待测样本液中的粒子因光照射所产生的至少两种散射光信号；
根据所述至少两种散射光信号，对所述待测血液样本中的红细胞和血小板进行分类。
一实施例中，所述稀释液能保持所述待测血液样本中的红细胞和血小板的原始形态。
一实施例中，所述至少两种散射光信号可以包括轴向光损失、前向散射光信号、中角散射光信号、高角散射光信号、侧向散射光信号及后向散射光信号中的至少两种。
一实施例中，所述轴向光损失、所述前向散射光信号、所述中角散射光信号、所述高角散射光信号、所述侧向散射光信号以及所述后向散射光信号的散射角度分别为0°~1°、1°~10°、10°~20°、20°~70°、70°~110°以及110°~160°。
一实施例中，所述至少两种散射光信号可以包括前向散射信号、中角散射信号、高角散射信号中的至少一种、尤其是至少两种。优选地，所述至少两种散射光信号可以包括前向散射信号和中角散射信号或者可以包括前向散射信号和高角散射信号。
一实施例中，所述光照射可以为偏振光照射，所述至少两种散射光信号包括所述样本液中的粒子因偏振光照射所产生的轴向光损失、前向散射光信号、中角散射光信号、高角散射光信号、侧向散射光信号以及后向散射光信号的特定偏振态信号中的至少两种。示例性地，所述至少两种散射光信号可以包括前向散射信号、中角散射信号、高角散射信号的特定偏振态信号中的至少一种、尤其是至少两种。优选地，所述至少两种散射光信号可以包括前向散射信号和中角散射信号的特定偏振态信号或者可以包括前向散射信号和高角散射信号的特定偏振态信号。
一实施例中，所述根据所述至少两种散射光信号，对所述待测样本中的红细胞和血小板进行分类的步骤可以包括：
根据所述至少两种散射光信号生成所述待测血液样本中的粒子的二维或三维散点图；
基于所述二维或三维散点图，对所述待测血液样本中的红细胞和血小板进行分类。
一实施例中，所述方法可以包括：根据所述至少两种散射光信号，求取所述待测血液样本中的红细胞和血小板的预定参数的检测结果。示例性地，所述预定参数可以包括红细胞计数、血小板计数、红细胞平均体积、血小板平均体积和红细胞体积分布宽度中的至少一个参数或其组合计算得到的参数。
一实施例中，所述方法还可以包括：输出所述待测血液样本的红细胞和血小板的分类结果和/或预定参数的检测结果。
另外，本发明实施例还提供了一种样本分析仪，包括：
采样装置，具有带吸移管嘴的吸移管并且具有驱动装置，该驱动装置用于驱动所述吸移管通过所述吸移管嘴定量吸取血液样本；
样本液制备装置，具有反应池和供液部，其中，所述反应池用于接收采样装置所吸取的血液样本，所述供液部将稀释液提供给反应池，从而由所述采样装置所吸取的血液样本与由所述供液部提供的稀释液在所述反应池中混合，以制备成待测样本液；
光学检测装置，具有光源、流动室以及集光器，其中，经稀释液处理后的待测样本液中的粒子可在所述流动室内流动，所述光源所发出的光照射所述流动室中的粒子以产生至少两种散射光信号，所述集光器用于收集所述至少两种散射光信号；
输送装置，用于将所述反应池中经稀释液处理后的待测样本液输送到所述光学检测装置中；
处理器，与所述采样装置、所述样本制备装置、所述光学检测装置以及所述输送装置通信连接并且用于：
指示所述样本制备装置制备含待测血液样本和稀释液的待测样本液；
指示所述输送装置将制备的测样本液输送至所述流动室；
从所述光学检测装置获取所述待测样本液中的粒子在所述流动室中因光照射所产生的至少两种散射光信号；
根据所述至少两种散射光信号对所述待测血液样本中的红细胞和血小板进行分类。
一实施例中，所述稀释液能保持所述血液样本中所述红细胞和所述血小板的原始形态。
示例性地，所述至少两种散射光信号可以包括轴向光损失、前向散射光信号、中角散射光信号、高角散射光信号、侧向散射光信号及后向散射光信号中的至少两种。
一实施例中，所述轴向光损失、所述前向散射光信号、所述中角散射光信号、所述高角散射光信号、所述侧向散射光信号以及所述后向散射光信号的散射角度分别为0°~1°、1°~10°、10°~20°、20°~70°、70°~110°以及110°~160°。
一实施例中，所述至少两种散射光信号可以包括前向散射信号、中角散射信号、高角散射信号中的至少一种、尤其是至少两种。优选地，所述至少两种散射光信号可以包括前向散射信号和中角散射信号或者可以包括前向散射信号和高角散射信号。
一实施例中，所述光源可以构成为发出偏振光的光源，所述至少两种散射光信号包括所述样本液中的粒子因偏振光照射所产生的轴向光损失、前向散射光信号、中角散射光信号、高角散射光信号、侧向散射光信号以及后向散射光信号的特定偏振态信号中的至少两种。
一实施例中，所述至少两种散射光信号包括前向散射信号、中角散射信号、高角散射信号的特定偏振态信号中的至少一种、尤其是至少两种。
一实施例中，所述处理器可以用于：
根据所述至少两种散射光信号生成所述待测血液样本中的粒子的二维或三维散点图；
基于所述二维或三维散点图，对所述待测血液样本中的红细胞和血小板进行分类。
一实施例中，所述处理器可以用于：根据所述至少两种散射光信号，求取所述待测血液样本中的红细胞和血小板的预定参数的检测结果。示例性地，所述预定参数可以包括红细胞计数、血小板计数、红细胞平均体积、血小板平均体积和红细胞体积分布宽度中的至少一个参数或其组合计算得到的参数。
一实施例中，所述样本分析仪还可以包括与所述处理器通信连接的输出装置，用于输出所述待测血液样本的红细胞和血小板的分类结果和/或预定参数的的检测结果。
 有益效果
根据本发明实施例的检测血液样本中的红细胞和血小板的方法及样本分析仪能够实现RBC和PLT的准确分类，尤其是在普通稀释液环境下实现RBC和PLT的准确分类。
 附图说明
图1为一种血细胞分析仪的光学检测装置的结构示意图；
图2为一实施例中的样本光学检测装置的结构示意图；
图3为一种用于说明第一前向散射光信号收集组件和第二前向散射光信号收集组件的结构的示意图；
图4为一种用于说明反射挡光组件的结构的示意图；
图5为一实施例中包括透镜组件的样本光学检测装置的结构示意图；
图6为一种用于透镜组件的结构的示意图；
图7为一实施例中包括挡直光阑的样本光学检测装置的结构示意图；
图8为一实施例中包括第三前向散射光信号收集组件的样本光学检测装置的结构示意图；
图9为一种用于说明第三前向散射光信号收集组件的结构的示意图；
图10为一实施例中包括侧向散射光信号收集组件和侧向荧光信号收集组件的样本光学检测装置的结构示意图；
图11为一种用于说明侧向散射光信号收集组件和侧向荧光信号收集组件的结构的示意图；
图12为一实施例中包括光源整形组件的样本光学检测装置的结构示意图；
图13为另一种实施例的样本光学检测装置的结构示意图；
图14为采用阻抗法检测RBC和PLT得到的直方图；
图15A为采用阻抗法检测正常样本的RBC和PLT得到的RBC直方图；
图15B为采用阻抗法检测正常样本的RBC和PLT得到的PLT直方图；
图16为采用阻抗法检测异常样本的RBC和PLT得到的PLT直方图；
图17为一实施例提供的样本分析仪的结构示意图；
图18为一实施例提供的检测血液样本中的红细胞和血小板的样本检测方法的示意性流程图；
图19为一实施例中各种散射光信号的散射角的示意图；
图20为一实施例提供的样本检测方法得到的前向散射光信号(FSC)与中角散射光信号(MAS)散点图；
图21为一实施例提供的样本检测方法得到的前向散射光信号(FSC)与高角散射光信号(WAS)散点图。
 本发明的实施方式
下面通过具体实施方式结合附图对本发明作进一步详细说明。其中不同实施方式中类似元件采用了相关联的类似的元件标号。在以下的实施方式中，很多细节描述是为了使得本申请能被更好的理解。然而，本领域技术人员可以毫不费力的认识到，其中部分特征在不同情况下是可以省略的，或者可以由其他元件、材料、方法所替代。在某些情况下，本申请相关的一些操作并没有在说明书中显示或者描述，这是为了避免本申请的核心部分被过多的描述所淹没，而对于本领域技术人员而言，详细描述这些相关操作并不是必要的，他们根据说明书中的描述以及本领域的一般技术知识即可完整了解相关操作。
另外，说明书中所描述的特点、操作或者特征可以以任意适当的方式结合形成各种实施方式。同时，方法描述中的各步骤或者动作也可以按照本领域技术人员所能显而易见的方式进行顺序调换或调整。因此，说明书和附图中的各种顺序只是为了清楚描述某一个实施例，并不意味着是必须的顺序，除非另有说明其中某个顺序是必须遵循的。
本文中为部件所编序号本身，例如“第一”、“第二”等，仅用于区分所描述的对象，不具有任何顺序或技术含义。而本申请所说“连接”、“联接”，如无特别说明，均包括直接和间接连接（联接）。
请参照图2，本发明一实施例中提供一种样本光学检测装置，该样本光学检测装置包括光源1、流动室6、第一前向散射光信号收集组件20和第二前向散射光信号收集组件30，下面具体说明。
流动室6用于供待测样本中的细胞逐个通过。例如将待测样本中的血细胞通过一些试剂溶解或染色后，采用鞘流技术，使得血细胞从流动室6中依次一个接一个地排队通过。图中Y轴方向为样本中血细胞运动的方向，需要说明的是，图中Y轴方向为垂直于纸面的方向。
光源1用于照射通过流动室6的细胞。一实施例中光源1为激光器，例如氦氖激光器或半导体激光器等。当光源1发出的光照射到流动室6中的细胞时会向周围产生散射。本发明一实施例中分别使用第一前向散射光信号收集组件20和第二前向散射光信号收集组件30收集第一角度范围的前向散射光和第二角度范围的前向散射光，下面具体说明。
第一前向散射光信号收集组件20用于收集第一角度范围的前向散射光，该前向散射光为光源1照射通过流动室6的细胞产生的且经反射后的光。请参照图3，一实施例中第一前向散射光信号收集组件20包括依次设置的第一角度限定光阑21、挡杂光光阑22和光电探测器23；第一角度限定光阑用于将经反射的前向散射光限定在第一角度范围并会聚于挡杂光光阑22，挡杂光光阑22则用于屏蔽第一角度范围的前向散射光的杂光，光电探测器23用于将收集的第一角度范围的前向散射光转换成电信号。在一实施例中，第一角度范围为低角度范围，例如第一角度范围为0到10度或1度到10度。
第二前向散射光信号收集组件30用于收集第二角度范围的前向散射光，该前向散射光为光源1照射通过流动室6的细胞产生的光。在一实施例中第二前向散射光信号收集组件30包括依次设置的挡杂光光阑31和光电探测器32，挡杂光光阑31用于屏蔽第二角度范围的前向散射光的杂光，光电探测器32用于将收集的第二角度范围的前向散射光转换成电信号。在一实施例中，第二角度范围为中角度范围，例如第二角度范围为10度到20度。在一实施例中，所述第一角度范围和第二角度范围为连续范围。
第一前向散射光信号收集组件20收集经反射后的第一角度范围的前向散射光，第二前向散射光信号收集组件30则直接收集或收集经折射或透射的前向散射光，通过这种光学结构的设计，可以压缩光路的纵向尺寸（图中Z轴方向），使得设计出来的样本光学检测装置小型化。
本发明一实施例中通过一反射挡光组件来实现对将第一角度范围和第二角度范围的前向散射光在空间上进行分离，一个被反射至第一前向散射光信号收集组件20，一个被允许通过到达第二前向散射光信号收集组件30，下面具体说明。
一实施例中的样本光学检测装置还包括反射挡光组件10，反射挡光组件10设置于光源1照射通过流动室6的细胞产生的前向散射光的光路上，用于将第一角度范围的前向散射光反射到第一前向散射光信号收集组件20，以及允许第二角度范围的前向散射光直接入射到所述第二前向散射光信号收集组件30。请参照图4（a）反射挡光组件10有许多种实现方式，例如一实施例中反射挡光组件10包括反射镜11，用于将第一角度范围的前向散射光反射到所述第一前向散射光信号收集组件20。一实施例中反射镜11可以为椭圆形，即反射镜11为椭圆孔径反射镜。反射镜11摆放的角度可以是45度，此时其镜面与图中Y轴平行，当然反射镜11摆放的角度也可以根据光路空间布局来调整，并不局限于只能45度。请参照图4（b），一实施例中反射挡光组件10括遮光条12，用于屏蔽除第二角度范围的前向散射光外的杂光。一实施例中反射镜11设置于该遮光条12上，例如当反射镜11为椭圆形时，反射镜11以长轴与遮光条12重合的方式设置于遮光条12上，如图4（c）所示。遮光条12在这里一是起到了反射镜11的支架作用，二是屏蔽除了第二角度范围的前向散射光外的杂光，这能够有效保证第二角度范围的前向散射光的信号质量。
为了进一步提升第一角度范围和第二角度范围的前向散射光的信号质量，请参照图5，在一实施例中样本光学检测装置还可以包括透镜组件13，透镜组件13设置于光源1照射通过流动室6的细胞产生的前向散射光的光路上，用于收集第一角度范围和第二角度范围的前向散射光，并出射给反射挡光组件10。例如一实施例中透镜组件13包括一个非球面透镜和一个球面透镜，或者，多个非球面透镜，或者，多个球面透镜，或者，一个非球面透镜和多个球面透镜。请参照图6，一具体实施例中透镜组件13可以由两个球面透镜14和15来构成的例子。一实施例中当透镜组件13包括球面透镜时，最靠近流动室6的球面透镜的有效数据孔径至少为0.34。透镜组件13的引入，可以保证系统像差被有效地校正，保证信号光斑能够被第一前向散射光信号收集组件20和第二前向散射光信号收集组件30这两个测量通道有效地收集。并且通过透镜组件13和反射挡光组件10的组合，可以有效保证第一角度范围的前向散射光的信号质量。
请参照图7，为了进一步提升第一角度范围和第二角度范围的前向散射光的信号质量，在一实施例中样本光学检测装置还可以包括挡直光阑16，设置于透镜组件13和反射挡光组件10之间。挡直光阑16有许多作用，例如挡直光阑16能够遮挡透镜组件13出射的前向散射光中的直射角度即0度的光——这可以防止信号饱和，和/或，能够将透镜组件13出射的前向散射光限定在第一角度范围和第二角度范围。
以上就是样本光学检测装置实现收集第一角度范围和第二角度范围的前向散射光的一些说明，在一些实施例中样本光学检测装置还能够实现收集第三角度范围的前向散射光。例如一实施例中第三角度范围为高角度范围，比如第三角度范围为20度到70度。一些实施例中第一角度范围、第二角度范围和第三角度范围为连续范围。下面具体说明样本光学检测装置如何实现收集第三角度范围的前向散射光。
考虑到样本光学检测装置中流动室6后面的光路空间比较紧张——例如其被设置有第一前向散射光信号收集组件20和第二前向散射光信号收集组件30，甚至被设置有透镜组件13等，因此如果利用光电探测器直接对前向散射光进行收集无法保证第三角度范围（例如高角度范围）的前向散射光可以被完全收集到，因此发明人考虑进行折射的光路结构来转折第三角度范围（例如高角度范围）的前向散射光，使得第三角度范围（例如高角度范围）的前向散射光都能够被有效收集。
因此请参照图8，一实施例中样本光学检测装置还包括第三前向散射光信号收集组件40，用于收集第三角度范围的前向散射光，该前向散射光为光源1照射通过流动室6的细胞产生的且经至少一次反射后的光。请参照图9，在一实施例中第三前向散射光信号收集组件40包括依次设置的反射镜41、第三角度范围孔径光阑42和光电探测器43；反射镜41用于将光源1照射通过流动室6的细胞经产生的第三角度范围的前向散射光反射至第三角度范围孔径光阑42，第三角度范围孔径光阑42用于前向散射光限定在第三角度范围，光电探测器43用于将收集的第三角度范围的前向散射光转换成电信号。需要说明的是，图9所示的样本光学检测装置只是为了示意第三前向散射光信号收集组件40所画的一个例子，这不代表包括第三前向散射光信号收集组件40的样本光学检测装置只能是图9所示的结构。
上述是样本光学检测装置对前向散射光的收集的一些说明，在一些实施例中本发明的样本光学检测装置还能够实现对侧向光——例如侧向散射光和/或侧向荧光的收集，下面结合图10和图11具体说明。
一实施例的样本光学检测装置还包括侧向散射光信号收集组件50，用于收集光源1照射通过流动室6的细胞产生的侧向散射光。一实施例中侧向散射光信号收集组件50包括依次设置的侧向小孔光阑51和光电探测器52，侧向小孔光阑51用于处理侧向散色光，光电探测器52则接收经侧向小孔光阑51处理后的侧向散色光并转换成电信号。
一实施例的样本光学检测装置还包括侧向荧光信号收集组件60，用于收集光源1照射通过流动室6的细胞产生的侧向荧光。一实施例中侧向荧光信号收集组件60包括依次设置的荧光小孔光阑61、滤光片62和光电探测器63，荧光小孔光阑61用于处理侧向荧光，处理后的侧向荧光经过滤光片62后到达光电探测器63，光电探测器63用于将侧向荧光转换成电信号。
在一实施例中可以先在流动室6的侧向光的路径上设置一收集透镜7来收集侧向光（包含有侧向散射光和侧向荧光），以及通过一二向色镜8将侧向散射光反射至侧向散射光信号收集组件50——例如二向色镜8将侧向散射光反射聚焦于侧向小孔光阑51，然后进入光电探测器52；侧向荧光则透过二向色镜8进入侧向荧光信号收集组件60——例如侧向荧光则透过二向色镜8聚焦于荧光小孔光阑61处，然后经过滤光片62后到达光电探测器63。
以上就样本光学检测装置对侧向光的收集的一些说明，一实施例中侧向散射光和侧向荧光可以为70度到110度的光。
需要说明的是，图10和图11是样本光学检测装置同时包括侧向散射光信号收集组件50和侧向荧光信号收集组件60的一个例子，在一些实施例中，样本光学检测装置也可以只包括侧向散射光信号收集组件50和侧向荧光信号收集组件60中的一者。
以上是光源1照射通过流动室6的细胞产生的前向散射光、侧向散射光和侧向荧光被相应的测量通道收集的一些说明，在一些实施例中光源1到流动室6之间光路也可以设置一些改善光信号的器件，例如请参照图12，一实施例中样本光学检测装置还可以包括光源整形组件9，用于对光源1发出的光束进行准直并使之会聚于通过流动室6的细胞。请参照图13，一实施例中光源整形组件9包括依次设置的准直透镜2、光隔离器3、第一柱面镜4和第二柱面镜5；准直透镜2用于对光源1发出的光束进行准直，光隔离器3用于抑制反馈光，防止后面的光学器件的反射的光进入光源1，对光源1产生影响；第一柱面镜4用于使光束在细胞通过的方向上会聚于所述流动室的中心，例如第一柱面镜4使光束在图中的Y轴方向上会聚于所述流动室的中心；第二柱面镜5用于将光束在垂直于细胞通过的方向上会聚，例如第二柱面镜5用于将光束在图中X轴方向上会聚，使得散射光都被照射到挡直光阑16内。本领域技术人员可以理解地，在一些情况下光源整形组件9中的器件不是必需的，例如光隔离器3和第二柱面镜5等。
综上所述，对样本光学检测装置中的五个测量通道都分别进行了说明，在具体实施时，可以根据需求引入这五个测量通道中的若干个。例如上面的图13就是一个五测量通道的样本光学检测装置的示意图。下面以图13为例子，再分别说明一下这五个测量通道的工作过程和原理。
图中样本细胞的流动方向（即通过流动室6的方向）为Y轴方向，即垂直于纸面的方向。从光源1发出的光线经过准直透镜2变为平等光束，经过光隔离器3，然后再经过第一柱面镜4的处理使得光束在样本细胞流动的方向即Y轴方向上会聚于所述流动室的中心，以照射通过流动室6的细胞。光隔离器3的主要作用是抑制反馈光——反馈光主要来自于光隔离器3后面的各光学元器件的反射——进入光源1，防止光源1输出功率产生波动，保证光学基线稳定。
第二柱面镜5、球面透镜14和15则会使得光束在X轴方向进行会聚，例如通过对第二柱面镜5、球面透镜14和15的设计，使得光束在X轴方向上的尺寸压缩到小于挡直光阑16的挡直条的横向尺寸，以保证直射光均能被挡直光阑16有效遮挡，防止信号饱和；需要说明的是，挡直光阑16的挡直条是沿Y轴方向的一直条，即挡直条的轴向是Y轴方向，横向是X轴方向。
当经过处理的血细胞逐个通过流动室6时，光源1的光束会最终照射到通过流动室6的细胞，产生第一角度范围的前向散射光（下面不妨以低角度的前向散射光为例）、第二角度范围的前向散射光（下面不妨以中角度的前向散射光为例）、第三角度范围的前向散射光（下面不妨以高角度的前向散射光为例）、侧向散射光和侧向荧光：
低角度的前向散射光和中角度的前向散射光由球面透镜14和15进行收集，之后低角度的前向散射光经反射挡光组件10（例如其椭圆孔径反射镜）反射至第一角度限定光阑21（例如低角度限定光阑）会聚于挡杂光光阑22处，然后进入到光电探测器23；而中角度的前向散射光则经反射挡光组件10（例如其遮光条12，遮光条12沿是X轴和Z轴组成的平面方向设置有，有效的遮挡方向为X轴方向），被有效地屏蔽了中角度的前向散射光的杂光，然后会聚于挡杂光光阑31，然后进入到光电探测器32；
高角度的前向散射光则经过反射镜41反射至第三角度范围孔径光阑42（例如高角度孔径光阑），然后进入光电探测器43；
侧向散射光和侧向荧光同时被收集透镜7收集，并在经过二向色镜8时进行空间分离，其中侧向散射光经过二向色镜8的前表面被反射聚焦于侧向小孔光阑51，然后进入光电探测器52；侧向荧光经透过二向色镜8后聚焦于荧光小孔光阑61，然后经过滤光片62后到达光电探测器63。
以上就是本发明的样本光学检测装置的一些说明。本发明利用透镜组件收集前向散射光中的低、中角前向散射光，并利用反射挡光组件对低角前向散射光和中角前向散射光进行空间分离，不仅保证了低角前向散射光和中角前向散射光的信号质量，同时减小了光学结构的纵向尺寸，十分有利于小型化；在紧凑的前向散射光的光路结构中，本发明利用反射镜对高角前向散射光进行收集，这可以替代传统通过侧向散射光信号对嗜酸粒细胞的分类；本发明在低角度前向散射光测量通道上，还增加了中角度前向散射光测量通道和高角度前向散射光测量通道，通过这三个测量通道可以在对一些细胞进行分类和计数，例如通过这三个测量通道可以在不使用荧光试剂或球形化试剂的条件下将血小板从红细胞中分类；本发明利用设置于流动室侧向光路上的收集透镜对侧向光进行消偏振和荧光信号收集，并通过二向色镜的波长选择对消偏振的侧向散射光和荧光信号进行分离。本发明实现了从0度或1度到110度的散射光的接收的光路结构，并且中间并没有出现被损失的角度。
接着参考图14至21详细描述本发明的样本检测方法和样本分析仪。
目前在血液分析仪中一般采用阻抗法或荧光光学法对血液样本中的RBC和PLT进行测量。
其中，阻抗法基于库尔特原理，使经过稀释的血样通过小孔，在小孔的两侧施加恒流源，每一个通过小孔的细胞都引起小孔内液体电阻抗的变化，从而生成电脉冲。获取对应的电脉冲，其幅度代表细胞的体积，从而生成RBC与PLT直方图，该直方图是一维信息，即只有细胞的体积信息，如图14所示。
在图14中，虚线左边为PLT直方图，虚线右边为RBC直方图。由于RBC粒子直径约为PLT的3倍，RBC粒子数量约为PLT的30倍，因此PLT直方图在RBC直方图的左边，且与横轴所围成的面积比RBC小得多。 也就是说，图14中虚线为RBC、PLT直方图的分界线。将RBC和PLT直方图分别做归一化处理，并单独画出来，就形成血细胞分析仪上常见的RBC直方图与PLT直方图，如图15A和15B所示，其中，图15A为采用阻抗法检测正常样本的RBC和PLT得到的RBC直方图；图15B为采用阻抗法检测正常样本的RBC和PLT得到的PLT直方图。
在正常样本的RBC和PLT直方图中，RBC峰与PLT峰之间存在明显的界限，通常将PLT直方图中PLT粒子主峰右侧的凹谷作为PLT与RBC的分界线，再分别对这两个直方图进行分析，得到PLT与RBC相关测量参数。但是对于某些异常样本，如低值PLT样本，其PLT直方图与RBC直方图交界处往往没有明显界限，如图16所示，低值PLT直方图呈现锯齿状，导致算法无法准确切割PLT与RBC直方图，进而不能获取准确的PLT测量结果。低值PLT的准确性是临床重点关注的血常规指标，这是阻抗法测量的主要缺点。
而荧光光学法可以克服此缺陷。荧光法基于流式细胞术。经过稀释和染色的样本在鞘流的挤压下，依次通过光学检测区。每个细胞经过激发光源的照射，在光学系统中获得前向散射信号(代表细胞体积)与荧光信号(代表细胞内核酸含量)，以此生成二维散点图，从而对RBC和PLT进行划分与计算。相对于阻抗法获得的一维直方图，光学法的二维散点图多了一个维度的信息，从而可以准确在二维散点图上对PLT和RBC进行划分。然而在荧光法中，需要荧光染料对细胞染色，而且需要使用特殊的稀释液对细胞进行球形化处理，所以使用成本较高，不利于光学法在临床的推广。
为了解决该问题，本发明提出一种仅利用散射光信息来检测血液血液样本中的红细胞和血小板的样本检测方法和样本分析仪，该样本检测方法和样本分析仪能够在普通稀释液环境下实现RBC和PLT的准确分类，尤其是能够实现在普通稀释液环境下对异常样本的RBC和PLT进行准确的分类。
首先参考图17详细说明本发明提供的样本分析仪。
如图17所示，该样本分析仪100至少包括采样装置（未示出）、样本液制备装置110、光学检测装置120、输送装置130和处理器140。
所述采样装置具有带吸移管嘴的吸移管并且具有驱动装置，该驱动装置用于驱动所述吸移管通过所述吸移管嘴定量吸取血液样本。进一步地，采样装置在吸取血液样本之后由其驱动装置驱动并移动至样本液制备装置110的反应池111，将所吸取的血液样本注入到该反应池111中。
所述样本液制备装置110具有至少一个反应池111并且具有供液部（未示出），其中，所述反应池111用于接收采样装置所吸取的血液样本，所述供液部将稀释液提供给反应池111，从而由所述采样装置所吸取的血液样本与由所述供液部提供的稀释液在所述反应池中反应，以制备成待检测血液样本。例如，供液部可以用于将适当的稀释液注入到反应池中，以对血液样本中的粒子进行处理，从而制备成待检测血液样本，用于后续检测。其中，所述稀释液选用血细胞分析仪必备的普通稀释液，以保持所述血液样本中所述红细胞和所述血小板的原始形态，无需使用使RBC、PLT球形化的特殊稀释液。例如，所述稀释液可以包括氯化钠，磷酸盐缓冲液以及防腐剂等成分，稀释液并不局限于某一种，可以根据需要进行选择，在此不再赘述。
所述光学检测装置120具有光源121、流动室122以及集光器123、124。光源121可以发射自然光或特定波段的光，并不局限于某一种。或者所述光源121还可以选用偏振光源，以发射特定偏振状态的偏振光。流动室122具有孔口1221，在样本液制备装置110中经稀释液处理后的待测样本液的粒子可在所述流动室122内流动，并逐个经过孔口1221。光源121所发出的光照射所述流动室122中的粒子以产生光学信号信息。集光器123、124用于收集所述光学信号信息。所述光学信号信息可以包括轴向光损失、前向散射光信号、中角散射光信号、高角散射光信号、侧向散射光信号及后向散射光信号中的至少两种，当选用偏振光进行照射时，所述光学信号信息包括所述轴向光损失、所述前向散射光信号、所述中角散射光信号、所述高角散射光信号、所述侧向散射光信号以及所述后向散射光信号的特定偏振态信号中的至少两种。即，光学检测装置120至少包括轴向光损失集光器、前向散射光信号集光器、中角散射光信号集光器、高角散射光信号集光器、侧向散射光信号集光器以及后向散射光信号集光器中的至少两种集光器。
在一种实施例中，所述集光器构成为光电探测器，如光电二极管或光电倍增管等。具体地，如图17所示，在流动室122中流动的血细胞发出的前向散射光通过聚光镜126和针孔127被光电二极管（前向散射光集光器）123接受，而侧向散射光通过聚光镜126、分色镜128、光学膜129和针孔127被光电倍增管（侧向散射光集光器）124接受。从各集光器123、124输出的光信号分别通过由放大器150进行放大和波形处理等模拟信号处理后被输送到处理器140中。
在本发明中，所述轴向光损失、所述前向散射光信号、所述中角散射光信号、所述高角散射光信号、所述侧向散射光信号以及所述后向散射光信号的散射角度分别为0°~1°、1°~10°、10°~20°、20°~70°、70°~110°以及110°~160°。其中，轴向光损失集光器、前向散射光信号集光器、中角散射光信号集光器、高角散射光信号集光器、侧向散射光信号集光器分别配置为接收上述散射角度的散射光信号。
优选地，所述光学检测装置120至少包括前向散射光信号集光器、中角散射光信号集光器和高角散射光信号集光器中的至少一种、尤其是至少两种，以接收前向散射信号、中角散射信号、高角散射信号中的至少一种、尤其是至少两种或前向散射信号、中角散射信号、高角散射信号的特定偏振态信号中的至少一种、尤其是至少两种散射光信号，以提高所述红细胞和血小板的检测结果的准确性。尤其是，所述光学检测装置120包括前向散射光信号集光器和中角散射光信号集光器或者包括前向散射光信号集光器和高角散射光信号集光器。
所述输送装置130用于将在反应池111中经稀释液处理后的待测血液样本输送到光学检测装置120中。
所述处理器140与采样装置、样本制备装置110、光学检测装置120以及输送装置通信130连接并且设置用于从所述光学检测装置120获取所述光学信号信息并且对所述光学信号信息进行处理，以求取待测血液样本中的粒子分类计数结果。处理器140可以具有未示出的A/D转换器，用于将由光学检测装置120提供的模拟信号转换为数字信号。具体地，处理器140用于实施以下还要详细说明的按照本发明的样本检测方法。
在本发明实施例中，上述处理器140可以为特定用途集成电路（Application Specific Integrated Circuit，ASIC）、数字信号处理器（Digital Signal Processor，DSP）、数字信号处理装置（Digital Signal Processing Device，DSPD）、可编程逻辑装置（ProgRAMmable Logic Device，PLD）、现场可编程门阵列（Field ProgRAMmable Gate Array，FPGA）、中央处理器（Central Processing Unit，CPU）、控制器、微控制器、微处理器中的至少一种。可以理解地，对于不同的设备，用于实现上述处理器功能的电子器件还可以为其它，本发明实施例不作具体限定。
接着参考图18至21对上述血液样本分析仪100检测血液样本中的红细胞和血小板的具体方法和原理进行详细的说明。
图18为一实施例提供的检测血液样本中的红细胞和血小板的样本检测方法。如图18所示，一种检测血液样本中的红细胞和血小板的样本检测方法200包括以下步骤：
步骤S210，制备含待测血液样本和稀释液的待测样本液。
例如，在该步骤中，例如在样本分析仪100的样本制备装置110中制备待测样本液，所述样本液制备装置110具有至少一个反应池111并且具有供液部（未示出），在处理器140的控制下，采样装置在吸取血液样本之后由其驱动装置驱动并移动至样本液制备装置110的反应池111，然后供液部将稀释液提供给反应池111，从而由所述采样装置所吸取的血液样本与由所述供液部提供的稀释液在所述反应池中反应，以制备成待检测血液样本。一方面，稀释液为待测血液样本提供了适宜的pH、电导率和渗透压，保证细胞的形态完整，不发生溶血作用；此外，稀释液还用于对上次检测样本残留的物质进行清洗，保证样本分析仪的采样针、管路和流动室的清洁，防止交叉污染；再一方面，样本分析仪暂时停止运行期间稀释剂要充盈在管路中间，防止外界灰尘等异物进入仪器，造成故障。其中，所述稀释液选用血细胞分析仪必备的普通稀释液，以保持所述血液样本中所述红细胞和所述血小板的原始形态，而不是使RBC、PLT球形化的特殊稀释液。
其中，在本发明的一实施例中，所述稀释液可以包括氯化钠，磷酸盐缓冲液以及防腐剂等成分，稀释液并不局限于某一种，可以根据需要进行选择，在此不再赘述。
步骤S220，在光学检测区对所述待测样本液进行光照射。
例如在该步骤中，在处理器140的控制下，样本分析仪100的输送装置130将在反应池111中经稀释液处理后的待测血液样本输送到光学检测装置120的流动池122中，经稀释液处理后的待测样本液的粒子可在所述流动室122内流动，并逐个经过孔口1221，光源121所发出的光照射所述流动室122中的粒子以产生光学信号信息。
步骤S230，收集所述待测样本液中的粒子因光照射所产生的至少两种散射光信号，即特定散射角度的散射光信号。
例如，在该步骤中，在处理器140的控制下，光学检测装置120将其输出的散射光信号输送到处理器140中，以便处理器140对所述散射光信号进行处理。在本发明的实施例中，所述散射光信号可以包括轴向光损失、前向散射光信号、中角散射光信号、高角散射光信号、侧向散射光信号及后向散射光信号中的至少两种。优选地，所述散射光信号包括前向散射信号、中角散射信号、高角散射信号中的至少一种、尤其是至少两种。更优选地，所述至少两种散射光信号包括前向散射信号和中角散射信号或者包括前向散射信号和高角散射信号。
其中，在本发明的实施例中所述散射角度指的是，以流动室中样本流与激发光束重合区域的中心为角度的顶点，以激发光束的传播方向为角度的第一条边，以在上述顶点的粒子发出的散射光的传播方向为角度的第二条边，由上述顶点和两条边所形成的角度。在没有相反说明的情况下，散射角度均参照该解释。
在此，本发明对所述散射光信号的散射角度进行如下定义，如图19所示，其中：所述轴向光损失的散射角度为：0°~1°；所述前向散射光信号的散射角度为：1°~10°；所述中角散射光信号的散射角度为：10°~20°；所述高角散射光信号的散射角度为：20°~70°；所述侧向散射光信号的散射角度为：70°~110°；并且所述后向散射光信号的散射角度为： 110°~160°。
在本发明的一实施例中，收集前向散射信号、中角散射信号、高角散射信号中的任意两种或全部用于后续对所述血液样本中的红细胞和血小板进行分类，选用上述三种散射光信号中的至少两种可以更有效和更加准确的对红细胞和血小板进行分类。具体地，例如，选用前向散射信号、中角散射信号、高角散射信号中至少两种获得到的红细胞和血小板的散点图中有更加清晰的红细胞和血小板分界线，由此进行分类的结果更加精准。
当然，需要说明的是，散射光信号的选择并不局限于上述示例，可以根据实际需要进行选择。
其中，所述光照射为可以为自然光或特定波段的光，还可以选用特定偏振状态的偏振光照射。当选用偏振光对所述待测样本液进行照射时，所述散射光信号为所述样本液中的粒子因偏振光照射而产生轴向光损失、前向散射光信号、中角散射光信号、高角散射光信号、侧向散射光信号以及后向散射光信号的特定偏振态信号。
类似地，在本发明的一实施例中，收集前向散射信号、中角散射信号、高角散射信号的特定偏振态信号中的任意两种或全部用于后续对所述血液样本中的红细胞和血小板进行分类，以获得更有效和更加准确的对红细胞和血小板进行分类。
步骤S240，根据所述至少两种散射光信号，对所述待测血液样本中的红细胞和血小板进行分类。
例如，处理器140从光学检测装置120接收所述散射光信号并进行处理，以获得所述待测血液样本中的红细胞和血小板的分类结果。
优选地，在该步骤S240中，对所述红细胞和所述血小板进行分类的步骤可以包括：根据所述至少两种散射光信号生成所述血液样本中的粒子的二维或三维散点图；基于所述二维或三维散点图，对所述血液样本中的红细胞和血小板进行分类。
在本发明的一实施例中，例如当待测样本液的PLT值＜30时，PLT直方图在右边界区呈现出锯齿状，如图16所示，难以划出与RBC区域相分离的分界线。但是采用本发明所述检测方法，可以生成如图20和图21所示的散点图。其中，图20为前向散射(FSC)光信号与中角散射(MAS) 光信号的散点图。图21为前向散射(FSC) 光信号与高角散射(WAS) 光信号的散点图。从图20和图21中可以看出PLT粒子团与RBC粒子团之间均存在明显界限，如图中虚线所示方向。因此，在阻抗法中无法区分的PLT与RBC粒子团能够在光散射散点图中得以区分，从而得到准确的RBC和PLT测量结果。
除了可以对红细胞和所述血小板进行分类以外，还可以根据所述至少两种散射光信号，求取所述红细胞和所述血小板的预定参数的检测结果。其中，所述预定参数可以包括红细胞计数、血小板计数、红细胞平均体积MCV、血小板平均体积MPV和红细胞体积分布宽度RDW中的至少一个参数以及由上述参数经过组合计算后得到的其他参数。
进一步地，所述方法200还可以包括输出所述待测血液样本的红细胞和血小板的分类计数结果和/或所述预定参数的检测结果。
根据本发明的检测血液样本中的红细胞和血小板的方法200及相应的样本分析仪能够实现RBC和PLT的准确分类，尤其是在普通稀释液环境下实现RBC和PLT的准确分类。在采用普通稀释液的情况下，血液样本中红细胞和血小板保持原始形态，例如红细胞保持双面凹的圆饼状，边缘较厚，而中间较薄，因此难以检测和区分。通过采用至少两种散射光以采集更多的细胞特征，能够弥补该缺陷，在使用普通稀释液的情况下仅利用散射光实现红细胞和血小板的区分。
以上应用了具体个例对本发明进行阐述，只是用于帮助理解本发明，并不用以限制本发明。对于本领域的一般技术人员，依据本发明的思想，可以对上述具体实施方式进行变化。 

Claims (43)

  1. 一种样本光学检测装置,其特征在于，包括：
    流动室，用于供待测样本中的细胞逐个通过；
    光源，用于照射通过所述流动室的细胞；
    第一前向散射光信号收集组件，用于收集第一角度范围的前向散射光，该前向散射光为所述光源照射通过流动室的细胞产生的且经反射后的光；
    第二前向散射光信号收集组件，用于直接收集第二角度范围的前向散射光，该前向散射光为所述光源照射通过流动室的细胞产生的光。
  2. 如权利要求1所述的样本光学检测装置，其特征在于，还包括反射挡光组件，设置于所述光源照射通过流动室的细胞产生的前向散射光的光路上，用于将第一角度范围的前向散射光反射到所述第一前向散射光信号收集组件，以及允许第二角度范围的前向散射光直接入射到所述第二前向散射光信号收集组件。
  3. 如权利要求2所述的样本光学检测装置，其特征在于，所述反射挡光组件包括反射镜，用于将第一角度范围的前向散射光反射到所述第一前向散射光信号收集组件。
  4. 如权利要求3所述的样本光学检测装置，其特征在于，所述反射镜为椭圆形。
  5. 如权利要求3或4所述的样本光学检测装置，其特征在于，所述反射挡光组件还包括遮光条，用于屏蔽除第二角度范围的前向散射光外的杂光；其中所述反射镜设置于该遮光条上。
  6. 如权利要求5所述的样本光学检测装置，其特征在于，所述反射镜以长轴与所述遮光条重合的方式设置于所述遮光条上。
  7. 如权利要求2至6中任一项所述的样本光学检测装置，其特征在于，还包括透镜组件，设置于所述光源照射通过流动室的细胞产生的前向散射光的光路上，用于收集第一角度范围和第二角度范围的前向散射光，并出射给所述反射挡光组件。
  8. 如权利要求7所述的样本光学检测装置，其特征在于，所述透镜组件包括一个非球面透镜和一个球面透镜，或者，多个非球面透镜，或者，多个球面透镜，或者，一个非球面透镜和多个球面透镜。
  9. 如权利要求8所述的样本光学检测装置，其特征在于，当所述透组件包括球面透镜时，最靠近流动室的球面透镜的有效数据孔径至少为0.34。
  10. 如权利要求7所述的样本光学检测装置，其特征在于，还包括挡直光阑，设置于所述透镜组件和反射挡光组件之间，用于遮挡透镜组件出射的前向散射光中的直射角度的光，和/或，将透镜组件出射的前向散射光限定在所述第一角度范围和第二角度范围。
  11. 如权利要求1所述的样本光学检测装置，其特征在于，所述第一前向散射光信号收集组件包括依次设置的第一角度限定光阑、挡杂光光阑和光电探测器；所述第一角度限定光阑用于将经反射的前向散射光限定在第一角度范围并会聚于所述挡杂光光阑，所述挡杂光光阑用于屏蔽第一角度范围的前向散射光的杂光，所述光电探测器用于将收集的第一角度范围的前向散射光转换成电信号。
  12. 如权利要求1所述的样本光学检测装置，其特征在于，所述第二前向散射光信号收集组件包括依次设置的挡杂光光阑和光电探测器，所述挡杂光光阑用于屏蔽第二角度范围的前向散射光的杂光，所述光电探测器用于将收集的第二角度范围的前向散射光转换成电信号。
  13. 如权利要求1所述的样本光学检测装置，其特征在于，还包括第三前向散射光信号收集组件，用于收集第三角度范围的前向散射光，该前向散射光为所述光源照射通过流动室的细胞产生的且经至少一次反射后的光。
  14. 如权利要求13所述的样本光学检测装置，其特征在于，所述第三前向散射光信号收集组件包括依次设置的反射镜、第三角度范围孔径光阑和光电探测器；所述反射镜用于将所述光源照射通过流动室的细胞经产生的第三角度范围的前向散射光反射至所述第三角度范围孔径光阑，所述第三角度范围孔径光阑用于前向散射光限定在第三角度范围，所述光电探测器用于将收集的第三角度范围的前向散射光转换成电信号。
  15. 如权利要求1至14中任一项所述的样本光学检测装置，其特征在于，所述第一角度范围为低角度范围，和/或，所述第二角度范围为中角度范围，和/或，所述第三角度范围为高角度范围。
  16. 如权利要求15所述的样本光学检测装置，其特征在于，所述第一角度范围和第二角度范围为连续范围。
  17. 如权利要求15所述的样本光学检测装置，其特征在于，所述第一角度范围、第二角度范围和第三角度范围为连续范围。
  18. 如权利要求15所述的样本光学检测装置，其特征在于，所述第一角度范围为0到10度或1度到10度；和/或，所述第二角度范围为10度到20度；和/或，所述第三角度范围为20度到70度。
  19. 如权利要求1所述的样本光学检测装置，其特征在于，还包括光源整形组件，用于对所述光源发出的光束进行准直并使之会聚于通过所述流动室的细胞。
  20. 如权利要求1至19中任一项所述的样本光学检测装置，其特征在于，所述光源整形组件包括依次设置的准直透镜和第一柱面镜，所述准直透镜用于对所述光源发出的光束进行准直，所述第一柱面镜用于使光束在细胞通过的方向上会聚于所述流动室的中心。
  21. 如权利要求20所述的样本光学检测装置，其特征在于，还包括设置于所述第一柱面镜的出射光路上的第二柱面镜，用于将光束在垂直于细胞通过的方向上会聚，使得散射光都被照射到所述挡直光阑内。
  22. 如权利要求20所述的样本光学检测装置，其特征在于，还包括设置于所述准直透镜和第一柱面镜之间的光隔离器，用于抑制反馈光。
  23. 如权利要求1所述的样本光学检测装置，其特征在于，还包括：
    侧向散射光信号收集组件，用于收集所述光源照射通过流动室的细胞产生的侧向散射光；和/或,
    侧向荧光信号收集组件，用于收集所述光源照射通过流动室的细胞产生的侧向荧光。
  24. 一种检测血液样本中的红细胞和血小板的样本检测方法，其特征在于，所述方法包括：
    制备含待测血液样本和稀释液的待测样本液；
    在光学检测区对所述待测样本液进行光照射；
    收集所述待测样本液中的粒子因光照射所产生的至少两种散射光信号，所述至少两种散射光信号中的至少一种选自前向散射信号、中角散射信号或高角散射信号；
    根据所述至少两种散射光信号，对所述待测血液样本中的红细胞和血小板进行分类。
  25. 根据权利要求24所述的方法，其特征在于，所述稀释液能保持所述待测血液样本中的红细胞和血小板的原始形态。
  26. 根据权利要求24或25所述的方法，其特征在于，所述至少两种散射光信号包括前向散射信号、中角散射信号和高角散射信号中的至少两种。
  27. 根据权利要求24至26中任一项所述的方法，其特征在于，所述前向散射光信号、所述中角散射光信号和所述高角散射光信号的散射角度分别为1°~10°、10°~20°以及20°~70°。
  28. 根据权利要求24至27中任一项所述的方法，其特征在于，所述至少两种散射光信号包括前向散射信号和中角散射信号或者包括前向散射信号和高角散射信号。
  29. 根据权利要求24或25所述的方法，其特征在于，所述光照射为偏振光照射，所述至少两种散射光信号包括所述样本液中的粒子因偏振光照射所产生的前向散射光信号、中角散射光信号和高角散射光信号的特定偏振态信号中的至少两种。
  30. 根据权利要求29所述的方法，其特征在于，所述至少两种散射光信号包括前向散射信号和中角散射信号的特定偏振态信号或者包括前向散射信号和高角散射信号的特定偏振态信号。
  31. 根据权利要求24至30中任一项所述的方法，其特征在于，所述根据所述至少两种散射光信号，对所述待测样本中的红细胞和血小板进行分类的步骤包括：
    根据所述至少两种散射光信号生成所述待测血液样本中的粒子的二维或三维散点图；
    基于所述二维或三维散点图，对所述待测血液样本中的红细胞和血小板进行分类。
  32. 根据权利要求24至31中任一项所述的方法，其特征在于，所述方法包括：根据所述至少两种散射光信号，求取所述待测血液样本中的红细胞和血小板的预定参数。
  33. 根据权利要求32所述的方法，其特征在于，所述预定参数包括红细胞计数、血小板计数、红细胞平均体积、血小板平均体积和红细胞体积分布宽度中的至少一个参数或其组合计算得到的参数。
  34. 根据权利要求33所述的方法，其特征在于，所述方法还包括：
    输出所述待测血液样本的红细胞和血小板的分类结果和/或预定参数。
  35. 一种样本分析仪，其特征在于，所述样本分析仪包括：
    采样装置，具有带吸移管嘴的吸移管并且具有驱动装置，该驱动装置用于驱动所述吸移管通过所述吸移管嘴定量吸取血液样本；
    样本液制备装置，具有反应池和供液部，其中，所述反应池用于接收采样装置所吸取的血液样本，所述供液部将稀释液提供给反应池，从而由所述采样装置所吸取的血液样本与由所述供液部提供的稀释液在所述反应池中混合，以制备成待测样本液；
    光学检测装置，具有光源、流动室以及集光器，其中，经稀释液处理后的待测样本液中的粒子可在所述流动室内流动，所述光源所发出的光照射所述流动室中的粒子以产生至少两种散射光信号，所述集光器用于收集所述至少两种散射光信号；
    输送装置，用于将所述反应池中经稀释液处理后的待测样本液输送到所述光学检测装置中；
    处理器，与所述采样装置、所述样本制备装置、所述光学检测装置以及所述输送装置通信连接并且用于：
    指示所述样本制备装置制备含待测血液样本和稀释液的待测样本液；
    指示所述输送装置将制备的测样本液输送至所述流动室；
    从所述光学检测装置获取所述待测样本液中的粒子在所述流动室中因光照射所产生的至少两种散射光信号，所述至少两种散射光信号中的至少一种选自前向散射信号、中角散射信号或高角散射信号；
    根据所述至少两种散射光信号对所述待测血液样本中的红细胞和血小板进行分类。
  36. 根据权利要求35所述的样本分析仪，其特征在于，所述稀释液能保持所述待测血液样本中的红细胞和血小板的原始形态。
  37. 根据权利要求35或36所述的样本分析仪，其特征在于，所述至少两种散射光信号包括前向散射光信号、中角散射光信号和高角散射光信号中的至少两种。
  38. 根据权利要求35至37中任一项所述的样本分析仪，其特征在于，所述前向散射光信号、所述中角散射光信号和所述高角散射光信号的散射角度分别为1°~10°、10°~20°和20°~70°。
  39. 根据权利要求35至38中任一项所述的样本分析仪，其特征在于，所述至少两种散射光信号包括前向散射信号和中角散射信号或者包括前向散射信号和高角散射信号。
  40. 根据权利要求35至39中任一项所述的样本分析仪，其特征在于，所述处理器用于：
    根据所述至少两种散射光信号生成所述待测血液样本中的粒子的二维或三维散点图；
    基于所述二维或三维散点图，对所述待测血液样本中的红细胞和血小板进行分类。
  41. 根据权利要求35至40中任一项所述的样本分析仪，其特征在于，所述处理器用于：根据所述至少两种散射光信号，求取所述待测血液样本中的红细胞和血小板的预定参数。
  42. 根据权利要求41所述的样本分析仪，其特征在于，所述预定参数包括红细胞计数、血小板计数、红细胞平均体积、血小板平均体积和红细胞体积分布宽度中的至少一个参数或其组合计算得到的参数。
  43. 根据权利要求42所述的样本分析仪，其特征在于，所述样本分析仪还包括与所述处理器通信连接的输出装置，用于输出所述待测血液样本的红细胞和血小板的分类结果和/或预定参数。
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