CN106198359A - Leukocyte differential counting instrument optical system using integrating sphere - Google Patents

Leukocyte differential counting instrument optical system using integrating sphere Download PDF

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CN106198359A
CN106198359A CN 201610521608 CN201610521608A CN106198359A CN 106198359 A CN106198359 A CN 106198359A CN 201610521608 CN201610521608 CN 201610521608 CN 201610521608 A CN201610521608 A CN 201610521608A CN 106198359 A CN106198359 A CN 106198359A
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integrating sphere
unit
light
optical system
aperture
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CN 201610521608
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Chinese (zh)
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邢博
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长春迪瑞医疗科技股份有限公司
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10Investigating individual particles
    • G01N15/14Electro-optical investigation, e.g. flow cytometers
    • G01N15/1434Electro-optical investigation, e.g. flow cytometers using an analyser being characterised by its optical arrangement
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10Investigating individual particles
    • G01N2015/1006Investigating individual particles for cytology

Abstract

The invention discloses a leukocyte differential counting instrument optical system using an integrating sphere. The optical system comprises an illumination unit, a sample unit, a signal collection unit and a signal processing unit and the above units are orderly arranged. The illumination unit is used for transversely gathering a light beam to the sample unit and longitudinally gathering the light beam to a bar-shaped diaphragm in the signal collection unit. The sample unit allows cells in the sample to pass through and produces diffusion lights at multiple angles under the light beam emitted by the illumination unit. The signal collection unit comprises an integrating sphere for acquiring diffusion lights at specified angles and carrying out beam splitting and a detector for acquiring the diffusion lights subjected to beam splitting. The signal processing unit is used for receiving diffusion light data transmitted by the detector and carrying out corresponding operation processing. The integrating sphere is used for beam splitting, effectively prevents a signal loss caused by crystal surface refraction-caused polarization effects and improves test accuracy. The detector is directly connected to the integrating sphere so that the whole optical system size is reduced.

Description

一种使用积分球的白细胞分类计数仪用光学系统 A method of using an integrating sphere optical leukocyte counter system

技术领域 FIELD

[0001]本发明涉及流式细胞仪技术领域,尤其涉及的是一种使用积分球的白细胞分类计数仪用光学系统。 [0001] The present invention relates to flow cytometry, and more particularly relates to a leukocyte classification using an integrating sphere optical system counter.

背景技术 Background technique

[0002]流式细胞技术是一种在液流系统中,快速测定单个细胞或细胞器的生物学性质,并把特定的细胞或细胞器从群体中加以分类收集的技术。 [0002] Flow cytometry is a system stream in rapid determination of the biological properties of single cells or organelles, and the particular cells or organelles technical classification to be collected from the population. 其特点是通过快速测定库尔特电阻、荧光、光散射和光吸收来定量测定细胞DNA含量、细胞体积、蛋白质含量、酶活性、细胞膜受体和表面抗原等许多重要参数。 Which is characterized by rapid determination Coulter resistance, fluorescence, light scattering and absorption of a number of important parameters to quantify DNA in cells, cell volume and protein content, enzyme activity, cell membrane receptors surface antigens, and the like. 根据这些参数将不同性质的细胞分开,以获得供生物学和医学研究用的纯细胞群体。 These parameters will be different according to the nature of the cells separated to obtain pure cell populations for biological and medical research purposes.

[0003]血细胞被相应试剂溶解或者染色后,采用鞘流技术,使血细胞从鞘流池中依次一个接一个的排队通过。 After [0003] Blood cells are lysed or staining agent appropriate, using the sheath flow technique, blood cells from the sheath flow cell one by one sequentially through the queue. 在细胞通过时,使用一个椭圆型的光斑照射细胞,光斑在细胞流向上的宽度应略大于细胞直径,以确保能够充分照明细胞,在垂直细胞流向上的长度应略大于鞘流宽度。 When the cells by using a cell elliptic spot is irradiated, the spot width in the flow direction should be slightly larger than the cell diameter of the cells, the cells to ensure that sufficient illumination, the vertical length of the cell in a flow of sheath flow should be slightly larger width. 它们会一个一个的通过激光照射区。 They will be a region irradiated by a laser. 当各种不同的细胞被激光照射时,其前向散射光,侧向散射光和侧向荧光的信号都是不同的。 When signals of different cells are irradiated with laser light, which is forward scattered light, side scattered light and side fluorescence are different. 利用这种不同,选择最能明显区分两种或者几种细胞的两路。 With this different, obvious that best distinguish between two or several two cells.

[0004]细胞经过光斑照射时会产生散射光。 [0004] The cells produce scattered light passes through the irradiation spot. 小角度散射光(2〜8度)的强弱主要表征了细胞的体积信息,大角度散射光(18〜24度)的强弱则主要表征了细胞内部颗粒复杂度的信息。 Small angle scattered light (2~8 degrees) the strength of the main volume of information characterizing the cells, large-angle light scattering (18~24 degrees) strength is mainly characterized by the complexity of the information cells within the particles. 由大小角度光强度构成二维坐标系,则每一个通过的细胞将在坐标系内表征为一个点。 Constituting dimensional coordinate system the size of the angle of the light intensity, each characterizing a cell through a point in the coordinate system. 多个细胞通过检测区域后,就形成了散点图。 After the cells through the plurality of detection areas, a scattergram is formed.

[0005]目前,请参考图1,细胞大小角度的光学信息在实际测量中一般被分成两路信号以单独测量,而这两路信号一般是应用分光棱镜100来分开的。 [0005] Currently, referring to FIG 1, the cell size of the angle of the optical information in the actual measurement is typically divided into two signals to measure separately and two signals are typically applied to separate the dichroic prism 100. 但是依据菲涅尔公式,光在电介质分界面上反射与折射会跟入射角度不同而不同,即材料对P光和S光的透射度不同,这就导致了光线通过分光棱镜I中的晶体时,所携带的光学信号不能完全的被传递,会对需要测量的信号造成损失。 However, based on Fresnel formula, electric light reflection and refraction with angle of incidence of the boundary surface of the medium varies, i.e., materials for light transmittance P S different light, which leads to the light passes through the dichroic prism crystals I , an optical signal can not be completely carried is transmitted, the signal will be measured loss.

[0006]因此,现有技术还有待于改进和发展。 [0006] Thus, the prior art has yet to be improved and developed.

发明内容 SUMMARY

[0007]鉴于上述现有技术的不足,本发明的目的在于提供一种使用积分球的白细胞分类计数仪用光学系统,旨在解决现有技术中使用分光棱镜对经细胞散射后的两路光线进行分光,所携带的光学信号不能完全的被传递,会对需要测量的信号造成损失,从而降低测试精度的缺陷。 [0007] In view of the deficiencies of the prior art, an object of the present invention is to provide a leukocyte classification using an integrating sphere optical system counter, designed to address the two light scattering cell through the use of the prior art dichroic prism disperses, the optical signal can not be completely carried is transmitted, the signal will be measured loss, thereby reducing the defect testing accuracy.

[0008]本发明的技术方案如下: [0008] The aspect of the present invention is as follows:

一种使用积分球的白细胞分类计数仪用光学系统,其中,包括依次设置的照明单元、样本单元、信号收集单元及信号处理单元;其中: A method of using an integrating sphere leukocyte counter optical system, comprising an illumination unit, a sample unit, a signal collecting unit and the signal processing unit are sequentially disposed; wherein:

所述照明单元,用于使光束横向汇聚至样本单元,并使光束纵向汇聚至信号收集单元中的条形光阑; The lighting unit is configured to converge the light beam transversely to the sample cell, and the longitudinal beam converged signal to stop strip collecting unit;

所述样本单元,用于使样本中的细胞依次通过,并在照明单元发出的光束下产生多种角度的散射光; The sample cell, a sample cell for sequentially through, and light scattering at multiple angles of light beams emitted from the lighting unit;

所述信号收集单元包括用于采集指定角度的散射光并进行分光的积分球,及用于采集分光后的散射光的探测器; Said signal collection unit comprises means for collecting prescribed angle scattered light and the spectral integrating sphere scattered light detector and means for collecting the beam splitter;

所述信号处理单元,用于接收探测器传输的散射光数据,并进行对应运算处理。 The signal processing unit, for receiving scattered light detector data transmission, and the corresponding arithmetic processing.

[0009]所述使用积分球的白细胞分类计数仪用光学系统,其中,所述照明单元包括: [0009] The use of integrating sphere leukocyte counter optical system, wherein the illumination unit comprises:

激光管; Laser tube;

光阑,用于将激光管发出的光束限定至指定光束孔径; Diaphragm for defining a light beam emitted from the laser tube to specify the beam aperture;

整形透镜,用于将限定光束孔径后的光束进行整形; Shaping lens for the light beam defining a beam shaping aperture;

第一柱面镜,用于使光束横向汇聚至样本单元; A first cylindrical lens for converging the light beam transversely to the sample cell;

第二柱面镜,用于使光束纵向汇聚至信号收集单元中的条形光阑。 A second cylindrical lens for converging the light beam to the signal longitudinal bar stop collecting unit.

[0010]所述使用积分球的白细胞分类计数仪用光学系统,其中,所述整形透镜的面型曲线为球面或非球面,或者所述整形透镜的面型横截面曲线为球面或非球面。 Shaped cross-sectional surface of [0010] using an integrating sphere the leukocyte counter optical system, wherein the plastic lens surface is spherical or aspherical curve, or a curve of the plastic lens is spherical or aspherical.

[0011 ]所述使用积分球的白细胞分类计数仪用光学系统,其中,所述激光管发出的光束经光阑限定光束孔径后,在样本单元中的细胞上形成椭圆形光斑,所述椭圆形光斑的长轴为80〜280um,短轴为9〜16um。 [0011] The use of the integrating sphere leukocyte counter optical system, wherein the light beam emitted by the laser tube defining a beam aperture diaphragm is formed in an elliptical spot on the sample unit cell, the elliptical the major axis of the spot 80~280um, a minor axis of 9~16um.

[0012]所述使用积分球的白细胞分类计数仪用光学系统,其中,所述样本单元包括鞘流池,所述鞘流池内部液体通路横截面为圆形或正方形。 [0012] The use of integrating sphere leukocyte counter optical system, wherein the sample cell comprises a sheath flow cell, the fluid sheath flow cell inside passage cross section is circular or square.

[0013]所述使用积分球的白细胞分类计数仪用光学系统,其中,所述积分球上设置有正对样本单元以接收散射光入射的入射孔,用于使入射光中角度为2-8°的散射光射出的小角度通光孔,及用于使入射光中角度为18-22°的散射光经过漫反射后射出的大角度通光孔。 [0013] The use of integrating sphere leukocyte counter optical system, wherein said integrating sphere is provided on a positive sample unit to receive the scattered light is incident into the aperture, the angle of incident light for 2-8 small ° scattering angle of light emitted through the light hole, and the angle of incident light for 18-22 ° light scattering after diffusely reflected light emitted from the through-hole large angle.

[0014]所述使用积分球的白细胞分类计数仪用光学系统,其中,所述小角度通光孔的中心轴与所述入射孔的中心轴在同一直线上,所述大角度通光孔的中心轴与所述小角度通光孔的中心轴垂直。 [0014] The integrating sphere leukocyte counter of the optical system, wherein the central axis of the small angle through the aperture center axis of the perforations along the same line, the large-angle light through hole the central axis through a small angle with the perpendicular central axis of the aperture.

[0015]所述使用积分球的白细胞分类计数仪用光学系统,其中,所述小角度通光孔的孔径2d*tan8° ;其中,d等于样本单元与入射孔之间的直线距离。 [0015] The integrating sphere leukocyte counter of the optical system, wherein the aperture angle of the light through the small hole 2d * tan8 °; wherein, d is equal to the sample cell and the linear distance between the perforations.

[0016]所述使用积分球的白细胞分类计数仪用光学系统,其中,所述探测器包括小角度探测器及大角度探测器,其中所述小角度通光孔的出口处设置有所述小角度探测器,所述大角度通光孔的出口处设置有所述大角度探测器。 [0016] The use of integrating sphere leukocyte counter optical system, wherein said detector comprises a small angle and large angle detector probe, wherein the small angle at the exit of light through the small hole provided an angle detector, a wide angle through the outlet aperture is provided with the large angle detector.

[0017]有益效果:本发明采用积分球代替分光棱镜进行分光,可以有效避免晶体表面折射而造成的偏振效应使信号损失,从而提高测试的准确度。 [0017] Advantageous Effects: The present invention uses an integrating sphere in place of the dichroic prism disperses, can effectively avoid the surface of the crystal and the polarization effect caused by refraction of the signal loss, thereby improving the accuracy of the test. 而且探测器与积分球直接相连,可以大大缩小整个光学系统的尺寸,减少收集镜的使用,更加节约成本。 And the detector was directly connected with the integrating sphere, greatly reduce the size of the entire optical system, reduce the use of the collection mirror, more cost.

附图说明 BRIEF DESCRIPTION

[0018]图1为现有技术中使用分光棱镜的白细胞分类计数仪用光学系统的示意图。 A schematic view of an optical system [0018] FIG. 1 is a prior art dichroic prism is used leukocyte counter used.

[0019]图2a为本发明所述使用积分球的白细胞分类计数仪用光学系统较佳实施例的结构框图。 [0019] FIG. 2a a block diagram of the present invention using an integrating sphere leukocyte counter of the preferred embodiment of the optical system.

[0020]图2b为本发明所述使用积分球的白细胞分类计数仪用光学系统较佳实施例的示意图。 [0020] FIG 2b of the present invention using an integrating sphere optical leukocyte counter system diagram of a preferred embodiment.

[0021]图3为本发明所述激光管发出的光束在样本单元中的细胞上形成椭圆形光斑的示意图。 [0021] FIG. 3 is a schematic elliptic beam spot is formed in the sample unit cell on the light beam emitted from the laser tube invention.

[0022]图4为本发明所述激光管发出的光束在第二柱面镜上形成光斑的示意图。 [0022] FIG. 4 is a schematic of a beam spot emitted from the laser tube is formed on the second cylindrical lens of the present invention.

具体实施方式 detailed description

[0023]本发明提供一种使用积分球的白细胞分类计数仪用光学系统,为使本发明的目的、技术方案及效果更加清楚、明确,以下对本发明进一步详细说明。 [0023] The present invention provides a method using an integrating sphere leukocyte counter optical system, for the purposes of the present invention, technical solutions and advantages clearer, explicit, the following detailed description of the present invention further. 应当理解,此处所描述的具体实施例仅仅用以解释本发明,并不用于限定本发明。 It should be understood that the specific embodiments described herein are only intended to illustrate the present invention and are not intended to limit the present invention.

[0024]请同时参见图2a和图2b,其中图2a为本发明所述使用积分球的白细胞分类计数仪用光学系统较佳实施例的结构框图,图2b为本发明所述使用积分球的白细胞分类计数仪用光学系统较佳实施例的示意图。 A block diagram [0024] Please also see Figures 2a and 2b, the present invention in which FIG. 2a using an integrating sphere leukocyte counter of the preferred embodiment of the optical system, the present invention FIG. 2b using integrating sphere leukocyte counter optical system diagram of a preferred embodiment. 所述使用积分球的白细胞分类计数仪用光学系统,包括依次设置的照明单元200、样本单元300、信号收集单元400及信号处理单元500;其中: The leukocyte classification using an integrating sphere optical system counter, includes an illumination unit 200 are sequentially disposed, a sample unit 300, a signal collecting unit 400 and the signal processing unit 500; wherein:

所述照明单元200,用于使光束横向汇聚至样本单元,并使光束纵向汇聚至信号收集单元中的条形光阑; The lighting unit 200 is configured to converge the light beam transversely to the sample cell, and the longitudinal beam converged signal to stop strip collecting unit;

所述样本单元300,用于使样本中的细胞依次通过,并在照明单元200发出的光束下产生多种角度的散射光; The sample cell 300 for the sample sequentially through the cells, and light scattering at multiple angles of light beams emitted from the lighting unit 200;

所述信号收集单元400包括用于采集指定角度的散射光并进行分光的积分球8,及用于米集分光后的散射光的探测器(图中未标出); The collection unit 400 includes a signal for acquiring the specified angle scattered light and the integrating sphere spectral 8, and a detector for scattered light collection spectral meter (not shown in the figure);

所述信号处理单元500,用于接收探测器传输的散射光数据,并进行对应运算处理。 The signal processing unit 500, for arithmetic processing corresponding to the received scattered light data transmission detector, and.

[0025]积分球8又称为光通球,是一个中空的完整球壳。 [0025] sphere 8, also known as luminous balls, is a hollow spherical shell intact. 内壁涂白色漫反射层,且球内壁各点漫射均匀。 Wall is coated with a white diffuse reflection layer, and an inner wall of ball points uniformly diffused. 光源在球壁上任意一点上产生的光照度是由多次反射光产生的光照度叠加而成的。 The light source of any point on the ball wall generated by the illumination light intensity is repeatedly reflected light of superposition. 其基本的工作原理为:光线由输入孔入射后,光线在球内部被均匀的反射及漫射,在球面上形成均匀的光强分布,因此输出孔所得到的光线为非常均匀的漫射光束。 The basic working principle is: the input aperture of light incident, reflected light is uniform and diffusion, a uniform light intensity distribution on the sphere inside the sphere, so the light output aperture was obtained as very uniform diffusion light beam . 而且入射光的入射角度、空间分布、以及极性都不会对输出的光强和均匀度造成影响。 And the incident angle of incident light, spatial distribution, and impact intensity and uniformity of the output will not polar. 同时因为光线经过积分球内部的均勾分布后才射出,其输出强度与输入强度比大约为:光输出孔面积/积分球内部的表面积。 At the same time because the light emitted through each hook after distribution inside the integrating sphere, the output intensity of the input intensity ratio of approximately: area of ​​the light output aperture area / point inside the sphere. 使用积分球可有效降低测量时因探测器上的入射光源不均匀分布或光束偏移所造成的微小误差。 Using an integrating sphere can reduce the measurement error due to minor source incident on the detector or uneven distribution of the beam displacement caused.

[0026]本发明的实施例中,采用积分球8代替分光棱镜进行分光,可以有效避免晶体表面折射而造成的偏振效应使信号损失,从而提高测试的准确度。 Example [0026] In the present invention, instead of using an integrating sphere 8 dichroic prism disperses, can effectively avoid the surface of the crystal and the polarization effect caused by refraction of the signal loss, thereby improving the accuracy of the test.

[0027] 优选的,如图2b所示,所述照明单元200包括: [0027] Preferably, as shown in FIG. 2b 200 of the lighting unit comprising:

激光管I; Laser tube I;

光阑2,用于将激光管I发出的光束限定至指定光束孔径; Diaphragm 2, the light beam for the laser tube to define a designated I emitted the beam aperture;

整形透镜3,用于将限定光束孔径后的光束进行整形; Shaping lens 3, the light beam for defining a beam shaping aperture;

第一柱面镜4,用于使光束横向汇聚至样本单元; First cylindrical lens 4 for converging the light beam transversely to the sample cell;

第二柱面镜5,用于使光束纵向汇聚至信号收集单元中的条形光阑。 5 a second cylindrical lens for converging the light beam to the signal longitudinal bar stop collecting unit.

[0028]在具体实施时,所述激光管I的光源为10w,波长635nm,其波长一定,发散角可以相同或者不同。 [0028] In a specific embodiment, the light source is a laser tube I is 10w, wavelength of 635nm, a wavelength constant, the divergence angles may be the same or different. 所述整形透镜3的面型曲线为球面或非球面,或者所述整形透镜3的面型横截面曲线为球面或非球面,用于对激光器I发出的光束进行准直、均匀、汇聚整形处理。 The shaping lens surface curve 3 is spherical or aspherical, plane or the curve-shaped cross-sectional shaping lens 3 is spherical or aspherical, a light beam emitted by the laser collimating I, uniform, converging shaping process . 所述激光管I发出的光束经光阑2限定光束孔径并通过第一柱面镜4后,在样本单元300中的细胞上形成椭圆形光斑,请参考图3,所述椭圆形光斑的长轴为80〜280um,短轴为9〜16um;最佳的,所述椭圆形光斑的长轴为110um,短轴为7um。 The light I emitted laser tube 2 via the aperture and the beam aperture defined by the first cylindrical lens 4, is formed in an elliptical spot on the sample unit cells 300, refer to FIG. 3, the long elliptical spot axis 80~280um, a minor axis 9~16um; optimal major axis of the elliptical spot is 110um, a minor axis of 7um. 所述激光管I在样本单元中的细胞上形成椭圆形光斑中,激光能量分布均匀,椭圆形光斑中心处与距椭圆形光斑中心5um处光能量密度差异不超过20%。 The laser tube is formed in the sample I on a cell unit in an elliptical spot, the laser energy distribution, the center of the elliptical spot of the light energy from the density difference of the elliptic beam spot center 5um not exceed 20%.

[0029]优选的,如图2b所示,所述样本单元300包括鞘流池6,所述鞘流池6内部液体通路横截面为圆形或正方形,当鞘流池6内部液体通路横截面为圆形时其直径小于25um,当鞘流池6内部液体通路横截面为正方形时其边长小于25um。 [0029] Preferably, shown in Figure 2b, the sample cell 300 includes a sheath flow cell 6, the sheath flow cell 6 inside the liquid passageway or square cross-section is circular, the sheath flow cell 6 when the inside of the liquid passage cross-section It is circular diameter less than 25um, the sheath flow cell 6 when the inside of the liquid passage cross section of a square side length of less than 25um. 所述样本单元300除了包括鞘流池6以外,还包括注样针以及其他管路。 The sample cell includes a sheath flow cell 300 except than 6, further comprising sample injection needle and another line. 其中只有鞘流池6在光路中,样本中的细胞会一个个的通过鞘流池6,照明单元200发出的光束会照射在鞘流池6的中央,并且在其中央聚焦。 6 wherein only the sheath flow cell in the light path, the sample cell will be one by one through the sheath flow cell 6, the center of the sheath flow cell 6 will illumination beam 200 emitted from the lighting unit, and is focused at the center. 在鞘流池6中,鞘液携裹着样本中的每一细胞依次流过。 In the sheath flow cell 6, each cell carrying wrapped sheath fluid sample flows through.

[0030]进一步的,所述积分球8与所述鞘流池6之间还设置有条形光阑7,所述激光管I发出的光束经光阑2并通过第二柱面镜5后,在条形光阑7上也形成光斑,该光斑也为椭圆形,这也是照明单元200在光路中的第二个焦点,使光束在横轴方向进行聚焦。 [0030] Further, the integrating sphere further disposed between the sheath flow cell 8 and the strip 6 with a diaphragm 7, the laser tube light I emitted by the cylindrical lens 5 after the second diaphragm 2 and , a spot is also formed on the stop strip 7, which is also an elliptical spot, the lighting unit 200 which is a second focal point in the optical path of the light beam focused on the horizontal axis.

[0031]优选的,所述积分球8上设置有正对样本单元300以接收散射光入射的入射孔,用于使入射光中角度为2-8°的散射光射出的小角度通光孔,及用于使入射光中角度为18-22°的散射光经过漫反射后射出的大角度通光孔。 [0031] Preferably, the unit is provided with a positive sample 300 for receiving the scattered light is incident into the aperture, the angle of incident light for scattering light emitted from the 2-8 ° angle through the small aperture 8 of the integrating sphere and for the angle of incident light to 18-22 ° scattered light emitted after diffuse reflection through a large angle of aperture.

[0032]其中,所述小角度通光孔的中心轴与所述入射孔的中心轴在同一直线上,所述大角度通光孔的中心轴与所述小角度通光孔的中心轴垂直。 [0032] wherein the central axis of light through a small angle with the entrance aperture hole central axis collinear with the central axis through a large angle perpendicular to the central axis of the aperture through the small angle of aperture . 所述小角度通光孔的孔径2d*tan8° ;其中,d等于样本单元300与入射孔之间的直线距离。 The small angle light through the aperture hole 2d * tan8 °; wherein, d is equal to the sample cell 300 and the linear distance between the perforations.

[0033]所述探测器包括小角度探测器10及大角度探测器9,其中所述小角度通光孔的出口处设置有所述小角度探测器10,所述大角度通光孔的出口处设置有所述大角度探测器9。 [0033] The detector 10 includes a small angle detector and the large angle detector 9, wherein the small angle through the outlet aperture is provided with a small angle to the detector 10, the large-angle light through the exit aperture there is provided at the large angle detector 9. 本发明中2-8°的散射光射出的小角度通光孔后,被小角度探测器10接收;18-22°的散射光由积分球8内壁进行反射,被大角度探测器9接收。 The present invention, after a small 2-8 ° scattering angle of light emitted from the light passing hole is smaller angle sensor 10 received; 18-22 ° scattered light is reflected by the integrating sphere inner wall 8, is a large angle detector 9 receives. 由于探测器与积分球8直接相连,可以大大缩小整个光学系统的尺寸,减少收集镜的使用,更加节约成本。 Since the detector 8 is directly connected to the integrating sphere, greatly reduce the size of the entire optical system, reduce the use of the collection mirror, more cost.

[0034]综上所述,本发明所述使用积分球的白细胞分类计数仪用光学系统,包括依次设置的照明单元、样本单元、信号收集单元及信号处理单元;照明单元,用于使光束横向汇聚至样本单元,并使光束纵向汇聚至信号收集单元中的条形光阑;样本单元,用于使样本中的细胞依次通过,并在照明单元发出的光束下产生多种角度的散射光;信号收集单元包括用于采集指定角度的散射光并进行分光的积分球,及用于采集分光后的散射光的探测器;信号处理单元,用于接收探测器传输的散射光数据,并进行对应运算处理。 [0034] In summary, the present invention is a lighting unit using an integrating sphere optical leukocyte counter system, comprising sequentially disposed, the sample unit, a signal collecting unit and the signal processing unit; lighting unit for transverse beam converge to the sample unit, and the longitudinal beam converged signal to stop strip collecting unit; sample cell, a sample cell for sequentially through, and light scattering at multiple angles of light beams emitted from the lighting unit; comprises a signal collecting unit for collecting the specified angle scattered light and the integrating sphere spectral, and a detector of scattered light collected after spectroscopic; a signal processing unit for receiving scattered light detector data transmission, and the corresponding the arithmetic processing. 本发明采用积分球进行分光,可有效避免晶体表面折射而造成的偏振效应使信号损失,从而提高测试准确度。 The present invention employs an integrating sphere spectral, can effectively prevent the surface of the crystal and the polarization effect caused by refraction of the signal loss, thereby improving the accuracy of the test. 而且探测器与积分球直接相连,可缩小整个光学系统的尺寸。 Further integrating sphere detector is directly connected to reduce the size of the entire optical system.

[0035]应当理解的是,本发明的应用不限于上述的举例,对本领域普通技术人员来说,可以根据上述说明加以改进或变换,所有这些改进和变换都应属于本发明所附权利要求的保护范围。 [0035] It should be appreciated that the present invention is applied is not limited to the above-described example, those of ordinary skill in the art, can be modified or converted according to the above description, all such modifications and variations shall fall within the appended claims of the invention protected range.

Claims (9)

  1. 1.一种使用积分球的白细胞分类计数仪用光学系统,其特征在于,包括依次设置的照明单元、样本单元、信号收集单元及信号处理单元;其中: 所述照明单元,用于使光束横向汇聚至样本单元,并使光束纵向汇聚至信号收集单元中的条形光阑; 所述样本单元,用于使样本中的细胞依次通过,并在照明单元发出的光束下产生多种角度的散射光; 所述信号收集单元包括用于采集指定角度的散射光并进行分光的积分球,及用于采集分光后的散射光的探测器; 所述信号处理单元,用于接收探测器传输的散射光数据,并进行对应运算处理。 1. A method of using an integrating sphere leukocyte counter, characterized in that the optical system comprising illumination means successively disposed, the sample unit, a signal collecting unit and the signal processing unit; wherein: said illumination means for transverse beam converge to the sample unit, and the longitudinal beam converged signal to stop strip collecting unit; the sample means for sequentially by cells in a sample, and produces a variety of angles in the beam emitted from the lighting unit scatter optical; comprises a signal collecting unit for collecting the specified angle scattered light and the integrating sphere spectral and scattered light detector for collecting the spectroscopic; the signal processing unit, for receiving a scatter transmission sensor light data, and the corresponding arithmetic processing.
  2. 2.根据权利要求1所述使用积分球的白细胞分类计数仪用光学系统,其特征在于,所述照明单元包括: 激光管; 光阑,用于将激光管发出的光束限定至指定光束孔径; 整形透镜,用于将限定光束孔径后的光束进行整形; 第一柱面镜,用于使光束横向汇聚至样本单元; 第二柱面镜,用于使光束纵向汇聚至信号收集单元中的条形光阑。 Using an integrating sphere according to claim 1, wherein said leukocyte classification counter optical system, wherein the illumination unit comprises: a laser tube; diaphragm for defining a light beam emitted from the laser tube to specify the beam aperture; shaping lens for the light beam defining a beam shaping aperture; a first cylindrical lens for converging the light beam transversely to the sample unit; and a second cylindrical lens for converging the light beam to the longitudinal direction of signal collecting unit shaped diaphragm.
  3. 3.根据权利要求2所述使用积分球的白细胞分类计数仪用光学系统,其特征在于,所述整形透镜的面型曲线为球面或非球面,或者所述整形透镜的面型横截面曲线为球面或非球面。 The use of the integrating sphere 2 leukocyte counter optical system as claimed in claim, wherein said shaping lens surface is spherical or aspherical curve, or the curved face-shaped cross-sectional shaping lens is spherical or aspherical.
  4. 4.根据权利要求2所述使用积分球的白细胞分类计数仪用光学系统,其特征在于,所述激光管发出的光束经光阑限定光束孔径后,在样本单元中的细胞上形成椭圆形光斑,所述椭圆形光斑的长轴为80〜280um,短轴为9〜16um。 The use of the integrating sphere 2 leukocyte counter optical system as claimed in claim, characterized in that, after the light beam emitted by the laser tube defining a beam aperture diaphragm is formed in an elliptical spot on the sample unit cell the major axis of the elliptical spot 80~280um, a minor axis of 9~16um.
  5. 5.根据权利要求1所述使用积分球的白细胞分类计数仪用光学系统,其特征在于,所述样本单元包括鞘流池,所述鞘流池内部液体通路横截面为圆形或正方形。 The use of an integrating sphere to the leukocyte counter optical system as claimed in claim, wherein said sample cell comprises a sheath flow cell, the fluid sheath flow cell inside passage cross section is circular or square.
  6. 6.根据权利要求1所述使用积分球的白细胞分类计数仪用光学系统,其特征在于,所述积分球上设置有正对样本单元以接收散射光入射的入射孔,用于使入射光中角度为2-8°的散射光射出的小角度通光孔,及用于使入射光中角度为18-22°的散射光经过漫反射后射出的大角度通光孔。 6. The use of an integrating sphere to the leukocyte counter optical system as claimed in claim, wherein said integrating sphere is provided with a positive sample unit to receive the scattered light is incident into the aperture, for causing the incident light small angle of 2-8 ° angle scattered light emitted through the light hole, and angle of incident light for the light scattering 18-22 ° after diffuse reflection through a large angle emitted from the aperture.
  7. 7.根据权利要求6所述使用积分球的白细胞分类计数仪用光学系统,其特征在于,所述小角度通光孔的中心轴与所述入射孔的中心轴在同一直线上,所述大角度通光孔的中心轴与所述小角度通光孔的中心轴垂直。 The integrating sphere leukocyte classification of the counter 6 with the optical system, characterized in that the central axis of the aperture through a small angle with the center axis of the entrance hole in the same straight line as claimed in claim, the large small angle with the central axis of the through aperture angle perpendicular to the axis through the center of the pupil.
  8. 8.根据权利要求6所述使用积分球的白细胞分类计数仪用光学系统,其特征在于,所述小角度通光孔的孔径2d*tan8° ;其中,d等于样本单元与入射孔之间的直线距离。 The use of the integrating sphere 6 leukocyte counter optical system as claimed in claim, wherein said small angle of aperture of the through aperture 2d * tan8 °; wherein, d is equal to the sample cell and between the entrance aperture Straight line distance.
  9. 9.根据权利要求6所述使用积分球的白细胞分类计数仪用光学系统,其特征在于,所述探测器包括小角度探测器及大角度探测器,其中所述小角度通光孔的出口处设置有所述小角度探测器,所述大角度通光孔的出口处设置有所述大角度探测器。 Leukocyte counter according to claim 6 using an integrating sphere claims, characterized in that the optical system, the detector comprises a small angle and large angle detectors detector, wherein the outlet through a small angle of aperture there is provided a small angle detector, a wide angle through the outlet aperture is provided with the large angle detector.
CN 201610521608 2016-07-05 2016-07-05 Leukocyte differential counting instrument optical system using integrating sphere CN106198359A (en)

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