JP2015190814A - particle measuring apparatus - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a particle measuring apparatus capable of increasing the amount of light to be detected while suppressing variations in light detection amount caused by mismatched optical parts.SOLUTION: A particle measuring apparatus 1 includes a flow cell 110, a photodetector 133, a first optical system 160 in between guiding lights emitted from particles toward the photodetector 133, and a second optical system 170 on the side opposite to the first optical system 160 with respect to the flow cell 110, reflecting lights emitted from the particles to the first optical system 160. The numerical aperture of the second optical system 170 for the flow cell 110 is set smaller than that of the first optical system 160 for the flow cell 110.

Description

  The present invention relates to a particle measuring apparatus for measuring particles by irradiating a flow containing particles such as blood cells with light.

  There is a particle measuring device that measures blood cells and particle components in a specimen using flow cytometry technology. In this type of particle measuring apparatus, it is preferable to detect as much light as possible from the particles. In the apparatus described in Patent Document 1, two pairs of optical filters and a photodetector are arranged with a flow channel interposed therebetween. Each optical filter is configured such that the wavelength of reflected light and the wavelength of transmitted light are opposite to each other. Thus, by reflecting a part of the light toward the photodetector on the opposite side by the optical filter, the light guided to the photodetector is enhanced.

US Patent No. 8031340

  In a configuration in which a part of the light generated from the flow cell is reflected and directed to the photodetector, the relative position between the reflected light and the optical system on the photodetector side when the optical component is displaced. Changes. Due to such a change in relative position, a part of the reflected light may be out of the light capturing range of the optical system on the photodetector side. For this reason, in the said structure, the light quantity which injects into a photodetector tends to fluctuate | variate greatly according to arrangement | positioning of an optical component. In order to eliminate fluctuations in the detected light amount, a complicated operation such as precise arrangement of optical components is required, which increases the burden on the operator.

  A particle measuring apparatus according to a main aspect of the present invention includes a flow cell for flowing particles, a light source for irradiating light to particles flowing in the flow cell, and particles flowing in the flow cell by being irradiated with light from the light source. A light detector for detecting light emitted from the light cell; a first optical system disposed between the flow cell and the light detector for guiding light emitted from the particles to the light detector; and A second optical system disposed on the opposite side of the first optical system and reflecting light emitted from the particles to the first optical system. Here, the numerical aperture of the second optical system for the flow cell is set smaller than the numerical aperture of the first optical system for the flow cell.

  According to the particle measuring apparatus according to this aspect, a part of the light emitted from the particles is guided to the photodetector by the first optical system. The other part of the light emitted from the particles is reflected by the second optical system and guided to the photodetector by the first optical system. Since the numerical aperture of the second optical system with respect to the flow cell is set to be smaller than the numerical aperture of the first optical system with respect to the flow cell, the spread of light reflected from the second optical system to the first optical system is increased. It becomes smaller than the range defined by the numerical aperture of the first optical system. For this reason, even if some variation occurs in the arrangement of the optical components, the light reflected from the second optical system to the first optical system does not deviate from the light capturing range of the first optical system. .

  ADVANTAGE OF THE INVENTION According to this invention, the particle | grain measuring apparatus which can increase a detected light quantity can be provided, suppressing the fluctuation | variation of the detected light quantity by the dispersion | variation in arrangement | positioning of an optical component.

  The effects and significance of the present invention will become more apparent from the following description of embodiments. However, the embodiment described below is merely an example when the present invention is implemented, and the present invention is not limited to the following embodiment.

It is a figure which shows typically the structure of the optical detection unit which concerns on Embodiment 1. FIG. FIG. 2A is a diagram schematically illustrating the configuration of the optical detection unit according to the first embodiment, and FIGS. 2B to 2E illustrate the flow cell, the beam stopper, the pinhole, and the second embodiment according to the first embodiment. It is a figure which shows the structure of a photodetector typically. It is a block diagram which shows the structure of the particle | grain measuring apparatus which concerns on Embodiment 1. FIG. FIGS. 4A and 4C are diagrams showing scattergrams according to the first embodiment, and FIG. 4B is a diagram showing absorption characteristics of hemoglobin contained in red blood cells. FIGS. 5A and 5B are schematic views showing the spread of reflected light according to the first embodiment. 6A to 6C are simulation results showing the spread of reflected light when the reflection-side member according to the comparative example is displaced in the X-axis direction. FIGS. 7A and 7B are simulation results showing the amount of light received by the photodetector when the reflection-side member according to the comparative example is displaced in the X-axis direction. 8A to 8C are simulation results showing the spread of reflected light when the reflection-side member according to the comparative example is displaced in the Y-axis direction. FIGS. 9A and 9B are simulation results showing the amount of light received by the photodetector when the reflection-side member according to the comparative example is displaced in the Y-axis direction. FIGS. 10A to 10C are simulation results showing the spread of reflected light when the reflection-side member according to the first embodiment is displaced in the X-axis direction. FIGS. 11A and 11B are simulation results showing the amount of light received by the photodetector when the reflection-side member according to the first embodiment is displaced in the X-axis direction. 12A to 12C are simulation results showing the spread of reflected light when the reflection-side member according to Embodiment 1 is displaced in the Y-axis direction. FIGS. 13A and 13B are simulation results showing the amount of light received by the photodetector when the reflection-side member according to the first embodiment is displaced in the Y-axis direction. FIG. 6 is a schematic diagram showing the spread of reflected light according to the second embodiment. FIGS. 15A to 15C are simulation results showing the spread of reflected light when the reflection-side member according to the second embodiment is displaced in the X-axis direction. FIGS. 16A and 16B are simulation results showing the amount of light received by the photodetector when the reflection-side member according to the second embodiment is displaced in the X-axis direction. FIGS. 17A to 17C are simulation results showing the spread of reflected light when the reflection-side member according to the second embodiment is displaced in the Y-axis direction. FIGS. 18A and 18B are simulation results showing the amount of light received by the photodetector when the reflection-side member according to the second embodiment is displaced in the Y-axis direction. FIG. 19A is a schematic diagram illustrating the spread of reflected light according to the first embodiment, and FIG. 19C is a schematic diagram illustrating the spread of reflected light according to the third embodiment. It is a figure which shows typically the structure of the optical detection unit which concerns on Embodiment 4. FIG. FIGS. 21A to 21C are schematic diagrams illustrating the spread of reflected light according to the modified example.

  In the following first to fourth embodiments, the present invention is applied to a device for detecting and analyzing blood related by detecting white blood cells, red blood cells, platelets, etc. contained in a blood sample and counting each blood cell. It is.

  <Embodiment 1>

  As shown in FIG. 1, the particle measuring apparatus 1 includes an optical detection unit 100. The optical detection unit 100 includes a flow cell 110, light sources 121 and 122, photodetectors 131 to 134, an optical system 140 that guides laser light emitted from the light sources 121 and 122 to the flow cell 110, and particles flowing through the flow cell 110. And an optical system 150 that guides the forward scattered light generated from the light to the corresponding photodetector. The optical detection unit 100 includes a first optical system 160 and a second optical system 170 that guide side scattered light and fluorescence generated from particles flowing through the flow cell 110 to the corresponding photodetectors, respectively. For convenience, FIG. 1 shows XYZ coordinate axes orthogonal to each other. FIG. 1 is a diagram of the optical detection unit 100 as viewed in the negative Y-axis direction.

  A measurement sample is sent into the flow cell 110 in a state of being wrapped in a sheath liquid. As shown in FIG. 2B, the flow cell 110 includes a sample nozzle 111, a sheath liquid supply port 112, a pore portion 113, and a waste liquid port 114. The sample nozzle 111 ejects the measurement sample upward toward the pore 113. A channel 115 through which the measurement sample flows is formed in the pore 113. The measurement sample contains particles such as blood cells.

  Returning to FIG. 1, the light source 121 is arranged so that the stacking direction of the semiconductor layers of the light emitting unit (not shown) matches the Z-axis direction. Accordingly, the spread angle of the laser light emitted from the light source 121 is maximized in the Z-axis direction and minimized in the Y-axis direction. The light source 121 emits light having a first wavelength in the negative X-axis direction. The light having the first wavelength is laser light having a wavelength of about 405 nm (hereinafter referred to as “blue laser light L1”). The outgoing optical axis of the light source 121 intersects the optical axis O of the optical system 140. The optical axis O is parallel to the Z axis.

  The light source 122 is arranged so that the stacking direction of the semiconductor layers of the light emitting unit (not shown) coincides with the X-axis direction. Therefore, the spread angle of the laser light emitted from the light source 121 is maximized in the X-axis direction and minimized in the Y-axis direction. The light source 122 emits light having the second wavelength in the positive Z-axis direction. The light of the second wavelength is laser light having a wavelength of about 640 nm (hereinafter referred to as “red laser light L2”). The outgoing optical axis of the light source 122 coincides with the optical axis O of the optical system 140.

  The optical system 140 includes collimator lenses 141 and 142, a dichroic mirror 143, a cylindrical lens 144, and a condenser lens 145.

  The collimator lens 141 converts the blue laser light L1 emitted from the light source 121 into parallel light. The collimator lens 142 converts the red laser light L2 emitted from the light source 122 into parallel light. The dichroic mirror 143 reflects the blue laser light L1 transmitted through the collimator lens 141 and transmits the red laser light L2 transmitted through the collimator lens 142. The dichroic mirror 143 is arranged so that the traveling direction of the blue laser light L1 reflected by the dichroic mirror 143 is slightly inclined from the Z-axis direction to the Y-axis positive direction, as shown in FIG.

  The cylindrical lens 144 converges the blue laser light L1 and the red laser light L2 that have passed through the dichroic mirror 143 only in the X-axis direction. The condenser lens 145 converges the blue laser light L1 and the red laser light L2 in the Y-axis direction and focuses them on the position of the flow path 115 of the flow cell 110. The condenser lens 145 converges the blue laser light L1 and the red laser light L2 in the X-axis direction so as to focus on the Z-axis negative side position of the flow path 115. As a result, the flow path 115 is irradiated with the blue laser light L1 and the red laser light L2 in a beam shape elongated in the X-axis direction, as shown in FIG.

  As shown in FIG. 2A, the blue laser light L1 reflected by the dichroic mirror 143 travels in a direction slightly tilted from the Z-axis direction to the Y-axis direction, so the irradiation position of the blue laser light L1 in the flow path 115 P1 is shifted in the positive Y-axis direction from the irradiation position P2 of the red laser beam L2. The irradiation position P2 of the red laser light L2 is on the optical axis O. When the blue laser light L1 is irradiated to the particle at the irradiation position P1, forward scattered light, side scattered light, and fluorescence are generated from the particle irradiated with the blue laser light L1. When the red laser light L2 is irradiated to the particles at the irradiation position P2, forward scattered light, side scattered light, and fluorescence are generated from the particles irradiated with the red laser light L2. The forward scattered light mainly travels from the flow cell 110 in the positive Z-axis direction. Side scattered light mainly travels from the flow cell 110 in the X-axis positive direction and the X-axis negative direction. The fluorescence mainly spreads around the flow cell 110.

  Hereinafter, the forward scattered light, the side scattered light, and the fluorescence generated by the blue laser light L1 are referred to as “blue forward scattered light L11”, “blue side scattered light L12”, and “blue fluorescence L13”, respectively. Forward scattered light, side scattered light, and fluorescence generated by the red laser light L2 are referred to as “red forward scattered light L21”, “red side scattered light L22”, and “red fluorescence L23”, respectively. The wavelengths of the blue forward scattered light L11 and the blue side scattered light L12 are substantially the same as those of the blue laser light L1, and the wavelengths of the red forward scattered light L21 and the red side scattered light L22 are substantially the same as those of the red laser light L2. The same. The wavelengths of the blue fluorescence L13 and the red fluorescence L23 are different from the wavelengths of the blue laser light L1 and the red laser light L2, and are determined by the reagents used. In this embodiment, the wavelength of the blue fluorescence L13 is 430 to 520 nm, and the wavelength of the red fluorescence L23 is a wavelength band exceeding 660 nm.

  As shown in FIG. 2B, the particles in the flow path 115 flow from the irradiation position P2 to the irradiation position P1. For this reason, there is a predetermined time lag from when the red laser light L2 is irradiated to the particle at the irradiation position P2 to when the same laser beam is irradiated to the same particle at the irradiation position P1. Therefore, the particle measuring apparatus 1 acquires such a time lag in advance, and based on the acquired time lag, for each particle, the detection signal of the light generated by the blue laser light L1 and the light generated by the red laser light L2. The detection signals are acquired in association with each other.

  Returning to FIG. 1, the optical system 150 includes a condenser lens 151, a beam stopper 152, and a pinhole 153.

  The condenser lens 151 is made of an achromatic lens and has a function of correcting chromatic aberration with respect to the blue forward scattered light L11 and the red forward scattered light L21. The condenser lens 151 condenses the blue forward scattered light L11 and the red forward scattered light L21 at the position of the pinhole 153. The condensing lens 151 is a part of the blue laser light L 1 and the red laser light L 2, and the blue laser light L 1 and the red laser light L 2 transmitted through the flow cell 110 without being irradiated on the particles are Focus on the position.

  As shown in FIG. 2A, the condensing lens 151 is arranged such that its optical axis is parallel to the Z axis and is shifted from the optical axis O of the optical system 140 in the positive direction of the Y axis. . As a result, the light beam passing through the center of the blue forward scattered light L11 passes through the condenser lens 151 and then proceeds in a direction inclined slightly from the Z-axis positive direction to the Y-axis negative direction. The light beam passing through the center of the red forward scattered light L21 passes through the condenser lens 151 and then travels in a direction inclined slightly from the Z-axis positive direction to the Y-axis positive direction.

  The beam stopper 152 passes most of the blue forward scattered light L11 and the red forward scattered light L21, and shields the blue laser light L1 and the red laser light L2 transmitted through the flow cell 110. The beam stopper 152 is configured by a thin plate member that does not transmit light. As shown in FIG. 2C, the beam stopper 152 includes semicircular openings 152a and 152b, and a light shielding portion 152c formed between the openings 152a and 152b. The beam stopper 152 is disposed so as to be positioned at the focal position in the X-axis direction of the blue laser light L1 and the red laser light L2. As a result, the blue laser light L1 and the red laser light L2 have a long beam shape in the Y-axis direction on the light shielding portion 152c, and are shielded by the light shielding portion 152c. Most of the blue forward scattered light L11 and the red forward scattered light L21 pass through the beam stopper 152 through the openings 152a and 152b.

  As shown in FIG. 2D, the pinhole 153 includes two holes 153a and 153b arranged in the Y-axis direction. The blue forward scattered light L11 is collected at the position of the hole 153a, and the red forward scattered light L21 is collected at the position of the hole 153b. The blue forward scattered light L11 and the red forward scattered light L21 pass through the holes 153a and 153b, respectively, and are received by the photodetector 131.

  As shown in FIG. 2E, the photodetector 131 is a photodiode, and includes two light receiving surfaces 131a and 131b arranged in the Y-axis direction. The light receiving surfaces 131a and 131b are arranged on the same plane. The blue forward scattered light L11 and the red forward scattered light L21 are applied to the light receiving surfaces 131a and 131b, respectively. The photodetector 131 outputs a signal based on the blue forward scattered light L11 (hereinafter referred to as “signal BFSC”) and a signal based on the red forward scattered light L21 (hereinafter referred to as “signal RFSC”).

  Returning to FIG. 1, the first optical system 160 includes a first lens 161, a first diaphragm member 162, a dichroic mirror 163, a condenser lens 164, a spectral filter 165, and a condenser lens 166. . The first lens 161 is arranged so that its optical axis coincides with a straight line parallel to the X axis passing through the flow path 115 of the flow cell 110.

  The first lens 161 is a collimator lens. Of the blue side scattered light L12, the red side scattered light L22, the blue fluorescence L13, and the red fluorescence L23, the blue fluorescence L13 and the red fluorescence L23 are substantially parallel light. Convert to The first diaphragm member 162 includes a circular opening 162a. The center of the opening 162a is on a straight line passing through the optical axis of the first lens 161 and parallel to the X axis. The first aperture member 162 defines the diameter of the light beam that passes through the first lens 161 and is guided to the photodetectors 132 and 133. The numerical aperture of the first optical system 160 with respect to the flow cell 110 is set by the first lens 161 and the first diaphragm member 162. In general, the numerical aperture is expressed by NA = n · sin θ.

  The dichroic mirror 163 reflects the red side scattered light L22 in the positive Z-axis direction and transmits the blue side scattered light L12, the blue fluorescence L13, and the red fluorescence L23. The transmission wavelength band of the dichroic mirror 163 is set so that the blue side scattered light L12, the blue fluorescence L13, and the red fluorescence L23 can be transmitted.

  The condensing lens 164 condenses the red side scattered light L22 reflected by the dichroic mirror 163. The photodetector 132 is a photodiode and includes a light receiving surface 132a. The red side scattered light L22 is collected on the light receiving surface 132a. The photodetector 132 outputs a signal (hereinafter referred to as “signal RSSC”) based on the red side scattered light L22.

  The spectral filter 165 absorbs the blue side scattered light L12 transmitted through the dichroic mirror 163, and transmits the blue fluorescence L13 and the red fluorescence L23 transmitted through the dichroic mirror 163. The absorption wavelength band of the spectral filter 165 is set so as to be able to absorb the blue side scattered light L12 and to transmit the blue fluorescence L13 and the red fluorescence L23.

  The condenser lens 166 collects the blue fluorescence L13 and the red fluorescence L23 that have passed through the spectral filter 165. The photodetector 133 is an avalanche photodiode and receives blue fluorescence L13 and red fluorescence L23. The photodetector 133 has a single light-receiving surface that covers both the condensing region of the blue fluorescence L13 and the condensing region of the red fluorescence L23. Instead, the photodetector 133 may include a light receiving surface that receives only the blue fluorescence L13 and a light receiving surface that receives only the red fluorescence L23. The photodetector 133 outputs a signal based on the blue fluorescence L13 (hereinafter referred to as “signal BFL”) and a signal based on the red fluorescence L23 (hereinafter referred to as “signal RFL”). In addition, the particle | grain measuring apparatus 1 acquires the signal based on the blue fluorescence L13 and the signal based on the red fluorescence L23 by driving any one of the light sources 121 and 122. FIG.

  The second optical system 170 includes a second lens 171, a second diaphragm member 172, and a reflection plate 173. The second lens 171 is arranged so that its optical axis coincides with a straight line parallel to the X axis passing through the flow path 115 of the flow cell 110.

  The second lens 171 is a collimator lens, and converts the blue fluorescence L13 out of the blue side scattered light L12, the red side scattered light L22, the blue fluorescence L13, and the red fluorescence L23 into parallel light. The second diaphragm member 172 includes a circular opening 172a. The center of the opening 172a is on a straight line passing through the optical axis of the second lens 171 and parallel to the X axis. The second diaphragm member 172 defines the diameter of the light beam that passes through the second lens 171. The numerical aperture of the second optical system 170 relative to the flow cell 110 is set by the second lens 171 and the second diaphragm member 172.

  Here, the first optical system 160 and the second optical system 170 are set so that the numerical aperture of the second optical system 170 with respect to the flow cell 110 is smaller than the numerical aperture of the first optical system 160 with respect to the flow cell 110. ing. Specifically, the first lens 161, the first diaphragm member 162, the second lens 171, and the second diaphragm member 172 are set so that the configuration and arrangement are in a desired state. Such setting will be described later with reference to FIG.

  A film structure that absorbs the red side scattered light L22 and the red fluorescent light L23, transmits the blue side scattered light L12, and reflects the blue fluorescent light L13 is formed on the incident surface of the reflecting plate 173. The photodetector 134 is a photodiode and includes a light receiving surface 134a. The blue side scattered light L12 is applied to the light receiving surface 134a. The photodetector 134 outputs a signal (hereinafter referred to as “signal BSSC”) based on the blue side scattered light L12.

  The blue fluorescence L13 reflected by the reflecting plate 173 travels in the positive direction of the X axis, passes through the second diaphragm member 172 and the second lens 171 again, and enters the flow cell 110. Then, the blue fluorescent light L13 incident on the flow cell 110 from the second lens 171 side passes through the flow cell 110, and similarly to the blue fluorescent light L13 generated in the X-axis positive direction from the particles in the flow path 115, the first optical system 160. Is guided to the photodetector 133.

  With reference to FIG. 3, the particle measuring apparatus 1 includes a measuring unit 10 and an information processing unit 20.

  The measurement unit 10 includes a measurement control unit 11, a sample suction unit 12, a sample preparation unit 13, and a detection unit 14. The measurement control unit 11 includes a memory 11a, and the detection unit 14 includes the optical detection unit 100 illustrated in FIG. The measurement control unit 11 drives each unit of the measurement unit 10 and receives a signal output from each unit. The measurement control unit 11 transmits and receives signals to and from the information processing unit 20. The information processing unit 20 includes a control unit 21, an output unit 22, an input unit 23, and a hard disk 24. The control unit 21 includes a memory 21a. The control unit 21 drives each unit of the information processing unit 20 and receives a signal output from each unit. In addition, the control unit 21 transmits and receives signals to and from the measurement unit 10. The output unit 22 is a display. The input unit 23 is a mouse or a keyboard.

  The operation of the particle measuring apparatus 1 is as follows.

  The sample suction unit 12 sucks a blood sample, which is peripheral blood collected from a patient, from the sample container, and sends the sucked blood sample to the sample preparation unit 13. The sample preparation unit 13 prepares a measurement sample used for measurement by mixing a reagent or the like with the specimen. The detection unit 14 outputs the signals BFSC, RFSC, BSSC, RSSC, BFL, and RFL output from the photodetectors 131 to 134 (see FIG. 1) to the measurement control unit 11. The measurement control unit 11 calculates a plurality of characteristic parameters such as a peak value, a width, and an area from the waveforms of the signals BFSC, RFSC, BSSC, RSSC, BFL, and RFL, respectively. The memory 11a stores a plurality of feature parameters in association with each particle. When the measurement is completed, the measurement control unit 11 transmits the characteristic parameter for each particle stored in the memory 11 a to the information processing unit 20.

  When the control unit 21 receives the characteristic parameter for each particle from the measurement unit 10, the control unit 21 performs analysis processing based on the computer program stored in the hard disk 24, and the hard disk 24 stores the analysis result. Upon receiving an analysis result display instruction via the input unit 23, the control unit 21 outputs a video signal including the analysis result to the output unit 22. The output unit 22 displays an image based on the video signal.

  Next, the particle classification performed in the analysis process will be described. In the analysis processing, the control unit 21 performs classification for each particle by combining a plurality of feature parameters obtained from the signals BFSC, RFSC, BSSC, RSSC, BFL, and RFL.

  Referring to FIG. 4A, when the control unit 21 classifies particles contained in the measurement sample into red blood cells, white blood cells, and platelets, the scattergram based on the blue forward scattered light L11 and the red forward scattered light L21. SG1 is used. The vertical axis and horizontal axis of the scattergram SG1 are the peak value of the signal RFSC and the peak value of the signal BFSC, respectively. The control unit 21 plots each particle in the measurement sample on the scattergram SG1, and sets regions A1 to A3 in the scattergram SG1. Thus, the control unit 21 classifies the particles included in the regions A1 to A3 into red blood cells, platelets, and white blood cells, respectively.

  As shown in FIG. 4A, in the scattergram SG1, a region A1 where red blood cells are distributed and a region A2 where white blood cells are distributed do not overlap each other. This is because red blood cells and white blood cells have different absorption levels of the blue laser light L1.

FIG. 4B shows the absorption coefficients of oxygenated hemoglobin (HbO ₂ ) and deoxygenated hemoglobin (Hb), respectively. In general, oxygenated hemoglobin and deoxygenated hemoglobin are present at a ratio of 3 to 1 in erythrocytes of venous blood. Therefore, the properties of oxygenated hemoglobin are dominant in the erythrocytes contained in the measurement sample. . As shown in FIG. 4B, in the wavelength range of 400 to 435 nm, the absorption coefficient of oxygenated hemoglobin is several steps larger than other wavelength bands. On the other hand, in the wavelength range of 610 to 750 nm, the absorption coefficient of oxygenated hemoglobin is several steps smaller than other wavelength bands. Therefore, red blood cells in the blood sample absorb blue laser light L1 more than red laser light L2. On the other hand, since other blood cells such as platelets and white blood cells do not contain hemoglobin, the other blood cells absorb blue laser light L1 and red laser light L2 to the same extent.

  Therefore, in red blood cells, the peak value of the blue forward scattered light L11 tends to be smaller than the peak value of the red forward scattered light L21, and in other blood cells, the peak value of the blue forward scattered light L11 and the peak of the red forward scattered light L21. The value tends to be comparable. Therefore, particles indicating red blood cells are distributed in the vicinity of the region A1, and particles indicating white blood cells are distributed in the vicinity of the region A3. Red blood cells are distributed along the distribution curve C1, and platelets and white blood cells are located on the distribution curve C2. From the above, according to the scattergram SG1 shown in FIG. 4A, red blood cells, white blood cells, and platelets can be classified well.

  With reference to FIG.4 (c), the control part 21 uses the scattergram SG2 based on the blue front scattered light L11 and the red front scattered light L21, when classifying the particle | grains contained in a measurement sample into three white blood cells. . The two axes of the scattergram SG2 are the same as the scattergram SG1 shown in FIG. The control unit 21 plots each particle in the measurement sample on the scattergram SG2, and sets regions A10, A3, A31 to A33 in the scattergram SG2. Particles included in region A10 are excluded. Thus, the control unit 21 classifies the particles included in the regions A31 to A33 into lymphocytes, monocytes, and granulocytes, respectively.

  Referring to FIG. 4D, when the control unit 21 classifies particles contained in the measurement sample into lymphocytes having surface antigen CD4 and lymphocytes not having surface antigen CD4, FIG. In addition to the scattergram SG2 shown in FIG. 5), a scattergram SG3 based on the red forward scattered light L21 and the red fluorescence L23 is used. The vertical axis and horizontal axis of the scattergram SG3 are the peak value of the signal RFSC and the peak value of the signal RFL, respectively. The control unit 21 plots each particle included in the region A33 of the scattergram SG2 on the scattergram SG3, and sets regions A41 and A42 in the scattergram SG3. Thus, the control unit 21 classifies the particles included in the regions A41 and A42 into lymphocytes each having the surface antigen CD4 and lymphocytes not having the surface antigen CD4.

  Referring to FIGS. 4E and 4F, when classifying the white blood cells contained in the measurement sample into three, the control unit 21 has a scattergram SG4 based on the blue forward scattered light L11 and the red forward scattered light L21. A scattergram SG5 based on the red forward scattered light L21 and the red side scattered light L22 can also be used. The two axes of the scattergram SG4 are the same as the scattergram SG2 shown in FIG. The vertical axis and the horizontal axis of the scattergram SG5 are values obtained by log display of the peak value of the signal RFSC and the peak value of the signal RSSC, respectively. The control unit 21 plots each particle in the measurement sample on the scattergram SG4, and sets regions A10 and A5 in the scattergram SG4. After excluding the particles included in the region A10, the control unit 21 plots each particle included in the region A5 on the scattergram SG5, and sets the regions A51 to A53 in the scattergram SG5. Thus, the control unit 21 classifies the particles included in the regions A51 to A53 into lymphocytes, monocytes, and granulocytes, respectively.

  In addition, the control part 21 can also use the scattergram based on the blue front scattered light L11 and the blue fluorescence L13, when classifying the particle contained in a measurement sample into malaria infection erythrocytes and other particles. When a predetermined fluorescent dye is used in the preparation of the measurement sample, red blood cells without nuclei are not stained, but DNA of malaria parasite is stained. Thereby, the control part 21 can classify malaria infection erythrocytes and other blood cells based on the scattergram which makes the peak value of the signal BFSC and the peak value of the signal BFL two axes.

  The measurement control unit 11 can also use a signal based on the blue side scattered light L12 when identifying particles included in the measurement particles. The measurement control unit 11 uses the signal based on the blue laser light L1 to identify finer particles than when using the signal based on the red laser light L2 having a longer wavelength than the blue laser light L1. Can do. Further, the blue laser light L1 transmitted through the flow cell 110 is likely to leak into the blue forward scattered light L11. For this reason, the measurement control part 11 can raise S / N ratio by using the signal based on the blue side scattered light L12 compared with the case where the signal based on the blue front scattered light L11 is used. Therefore, the measurement control unit 11 can identify particles with high accuracy by using a signal based on the blue side scattered light L12.

  Next, the numerical aperture of the first optical system 160 and the numerical aperture of the second optical system 170 will be described.

  As shown in FIG. 5A, the first lens 161 and the second lens 171 are configured so that the focal length is f1. The opening 162a of the first diaphragm member 162 is set to have a diameter of φ1, and the opening 172a of the second diaphragm member 172 is set to have a diameter of φ2 smaller than φ1. That is, the aperture diameter of the first aperture member 162 is φ1, and the aperture diameter of the second aperture member 172 is φ2. At this time, on the positive X-axis side of the flow path 115, the light from the flow path 115 passes through the first diaphragm member 162 and is taken into the first optical system 160 in the range of the angle θ1. In addition, on the X axis negative side of the flow path 115, the light from the flow path 115 passes through the second diaphragm member 172 and is taken into the second optical system 170 in a range of an angle θ2 smaller than the angle θ1. As described above, when the first lens 161, the first diaphragm member 162, the second lens 171, and the second diaphragm member 172 are set, the numerical aperture of the second optical system 170 with respect to the flow cell 110 is set to the flow cell 110. Is smaller than the numerical aperture of the first optical system 160.

  Note that the numerical aperture of the first optical system 160 with respect to the flow cell 110 is defined by light taken from the flow cell 110 and taken into the first optical system 160 and guided to the corresponding photodetector. . That is, the numerical aperture of the first optical system 160 with respect to the flow cell 110 is defined by the divergence angle θ1 when this light travels from the flow cell 110 in the positive X-axis direction. Further, the numerical aperture of the second optical system 170 with respect to the flow cell 110 is light that is taken in and reflected by the second optical system 170 and guided to the first optical system 160 among the light generated from the flow cell 110. It is prescribed by. That is, the numerical aperture of the second optical system 170 relative to the flow cell 110 is defined by the divergence angle θ2 when this light travels from the flow cell 110 in the negative X-axis direction.

  For example, as shown in FIG. 5B, when the second diaphragm member 172 is installed away from the second lens 171 in the negative direction of the X axis, the first optical system is reflected by the second optical system 170. The range of light guided to 160 is the range of angle θ2. In this way, when the transmitted light is parallel light, the numerical aperture of the second optical system 170 relative to the flow cell 110 is the same as that of FIG. 5A even if the position of the second diaphragm member 172 in the X-axis direction changes. As in the case of, it is defined by an angle θ2 corresponding to the range of light guided to the first optical system 160.

  In FIG. 5A, a light flux of light reflected by the reflecting plate 173 (hereinafter referred to as “reflected light”) is indicated by a broken line, and the first lens 161 does not pass through the second optical system 170. The luminous flux of light incident on is indicated by a solid line. The diameter of the light flux of the reflected light is restricted to φ2 by the second diaphragm member 172. Therefore, the reflected light enters the first diaphragm member 162 from the flow path 115 at an angle θ2 smaller than the angle θ1, and the reflected light passes through the inside of the opening 162a when passing through the first diaphragm member 162. become. As a result, even if there is some variation in the arrangement of the optical components included in the first optical system 160 and the second optical system 170, the reflected light is unlikely to come off from the opening 162a of the first diaphragm member 162. . Therefore, the amount of reflected light incident on the photodetector 133 does not change greatly due to variations in the arrangement of optical components.

  Next, simulation results performed by the present inventor will be described. The present inventor assumes a case where the second lens 171 and the second diaphragm member 172 (hereinafter referred to as “reflection-side member”) are misaligned as an example of the case where the arrangement of the optical components varies. did. In this case, the present inventor examined how the amount of received light detected by the photodetector 133 changes.

  First, the simulation results of the comparative example will be described with reference to FIGS. 6 (a) to 13 (b). In the comparative example, the diameter φ2 of the opening 172a of the second diaphragm member 172 is the same as the diameter φ1 of the opening 162a of the first diaphragm member 162. That is, in the comparative example, the numerical aperture of the second optical system 170 for the flow cell 110 is the same as the numerical aperture of the first optical system 160 for the flow cell 110.

  As shown in FIG. 6B, when the reflection side member is not displaced, the reflected light traveling in the positive direction of the X axis is not shielded by the second aperture member 172 and the first aperture member 162. On the other hand, as shown in FIG. 6A, when the reflection side member is shifted by −0.3 mm in the X-axis direction, the peripheral portion of the reflected light is shielded by the second diaphragm member 172. Further, as shown in FIG. 6C, when the reflection side member is shifted by +0.3 mm in the X-axis direction, the peripheral portion of the reflected light is shielded by the first diaphragm member 162.

  As shown in FIG. 7A, when the reflection side member is displaced in the X-axis direction, the amount of received light detected by the photodetector 133 decreases according to the amount of deviation. FIG. 7B is a graph obtained by normalizing the graph of FIG. 7A with the maximum amount of received light. As shown in FIG. 7B, the amount of received light when the amount of deviation in the X-axis direction is +0.3 mm is reduced by about 43% compared to the maximum amount of received light.

  As shown in FIG. 8B, when the reflection side member is not displaced, the reflected light traveling in the positive X-axis direction is not shielded by the second diaphragm member 172 and the first diaphragm member 162. On the other hand, as shown in FIGS. 8A and 8C, when the reflection side member is shifted by 0.3 mm in the Y-axis direction, the peripheral portions of the reflected light are the second diaphragm member 172 and the first diaphragm member 162. Is shielded from light.

  As shown in FIG. 9A, when the reflection-side member is displaced in the Y-axis direction, the amount of received light detected by the photodetector 133 decreases according to the amount of deviation. FIG. 9B is a graph obtained by normalizing the graph of FIG. 9A with the maximum amount of received light. As shown in FIG. 9B, the amount of received light when the amount of deviation in the Y-axis direction is 0.15 mm and 0.3 mm is about 30% and about 56%, respectively, compared to the maximum amount of received light. It will decrease.

  The behavior of the reflected light when the reflection side member is displaced in the Y-axis direction and the behavior of the reflected light when the reflection side member is displaced in the Z-axis direction are the same in principle. Therefore, the simulation results when the reflection-side member is displaced in the Z-axis direction are the same as those in FIGS. 8A to 9B.

  Next, the simulation result of Embodiment 1 is demonstrated.

  As shown in FIG. 10B, when the reflection side member is not displaced, the reflected light traveling in the positive X-axis direction is not shielded by the second diaphragm member 172 and the first diaphragm member 162. On the other hand, as shown in FIGS. 10A and 10B, when the reflection-side member is shifted by 0.3 mm in the X-axis direction, the peripheral portion of the reflected light is slightly shielded by the second diaphragm member 172. . However, the reflected light is not substantially shielded by the first diaphragm member 162. The reason for this is that, as described with reference to FIG. 5A, the range of reflected light when passing through the first diaphragm member 162 is narrower than the opening 162a, and there is a gap between the reflected light and the opening 162a. Because there is.

  As shown in FIG. 11A, when the reflection-side member is displaced in the X-axis direction, the amount of received light detected by the photodetector 133 slightly decreases according to the amount of deviation. FIG. 11B is a graph obtained by normalizing the graph of FIG. 11A with the maximum amount of received light. As shown in FIG. 11 (b), the amount of received light when the amount of deviation in the X-axis direction is +0.3 mm is suppressed to a decrease of about 20% compared to the maximum amount of received light. On the other hand, in the comparative example, as described above, the amount of received light when the amount of deviation in the X-axis direction is +0.3 mm is reduced by about 43% compared to the maximum amount of received light. Therefore, in Embodiment 1, it can be seen that even when the reflection-side member is displaced in the X-axis direction, the rate of decrease in the amount of received light can be suppressed as compared with the comparative example.

  As shown in FIG. 12B, when the reflection side member is not displaced, the reflected light traveling in the positive direction of the X axis is not shielded by the second aperture member 172 and the first aperture member 162. On the other hand, as shown in FIGS. 12A and 12C, when the reflection side member is shifted by 0.3 mm in the Y-axis direction, the peripheral portions of the reflected light are the second diaphragm member 172 and the first diaphragm member 162. Is shielded from light.

  As shown in FIG. 13A, when the reflection side member is displaced in the Y-axis direction, the amount of received light detected by the photodetector 133 decreases according to the amount of deviation. FIG. 13B is a graph obtained by normalizing the graph of FIG. 13A with the maximum amount of received light. As shown in FIG. 13B, the amount of received light when the amount of deviation in the Y-axis direction is 0.15 mm and 0.3 mm is about 15% and about 46%, respectively, compared to the maximum amount of received light. Reduced. On the other hand, in the comparative example, as described above, the received light amount when the amount of deviation in the Y-axis direction is 0.15 mm and 0.3 mm is about 30% and about 30%, respectively, compared to the maximum received light amount. Reduced by 56%. Therefore, in Embodiment 1, it can be seen that even when the reflection-side member is displaced in the Y-axis direction, the reduction rate of the received light amount can be suppressed lower than that in the comparative example. In the first embodiment, when the reflection-side member is displaced in the Y-axis direction, the amount of received light is gradually reduced as compared with the comparative example. In particular, it can be seen that in the range where the amount of deviation in the Y-axis direction is small, the reduction rate of the amount of received light is kept low.

  Note that the simulation results when the reflection-side member is displaced in the Z-axis direction are also the same as in FIGS. 12 (a) to 13 (b). Therefore, it can be seen that even when the reflection-side member is displaced in the Z-axis direction, the decrease in the amount of received light can be suppressed lower than in the comparative example.

  As described above, according to the first embodiment, the numerical aperture of the second optical system 170 with respect to the flow cell 110 is set smaller than the numerical aperture of the first optical system 160 with respect to the flow cell 110. For this reason, the spread angle of the reflected light is smaller than the angle range defined by the numerical aperture of the first optical system 160. That is, the reflected light is incident on the first diaphragm member 162 at an angle θ2 smaller than the angle θ1, and a gap is generated between the reflected light and the opening 162a. As a result, even if there is some variation in the arrangement of the optical components such as the first lens 161, the first diaphragm member 162, the second lens 171, and the second diaphragm member 172, the reflected light is not reflected in the first optical element. It becomes difficult to deviate from the light capturing range of the system 160. Therefore, even if the optical components vary, the light amount fluctuation of the reflected light detected by the photodetector 133 can be suppressed. Specifically, the light amount fluctuation of the blue fluorescence L13 detected by the photodetector 133 is suppressed.

  According to the first embodiment, the reflecting plate 173 reflects the blue fluorescence L13 generated in the X-axis negative direction from the particles in the flow path 115. Thereby, the photodetector 133 can receive blue fluorescence L13 generated in the X-axis positive direction from the particles in the flow path 115 and blue fluorescence L13 generated in the X-axis negative direction from the particles in the flow path 115. . Therefore, it is possible to increase the amount of light detected by the photodetector 133 with respect to the blue fluorescence L13.

  According to the first embodiment, the numerical aperture of the first optical system 160 with respect to the flow cell 110 is set as the numerical aperture of the first lens 161 by combining the first lens 161 with the first diaphragm member 162. The numerical aperture of the second optical system 170 is set as the numerical aperture of the second lens 171 by combining the second lens 171 with the second diaphragm member 172. Therefore, by adjusting the aperture diameter φ1 of the first aperture member 162, the numerical aperture of the first optical system 160 can be easily set, and the aperture diameter φ2 of the second aperture member 172 is adjusted. Thus, the numerical aperture of the second optical system 170 can be easily set. Furthermore, since the numerical apertures of the first optical system 160 and the second optical system 170 can be set by the aperture diameters of the first diaphragm member 162 and the second diaphragm member 172, the first lens 161 and the second lens 171 are: Lenses having the same focal length f1 can be used. Thereby, the cost concerning the 1st lens 161 and the 2nd lens 171 can be held down.

  According to the first embodiment, the reflecting plate 173 transmits the blue side scattered light L12 and guides it to the photodetector 134. This eliminates the need for the first optical system 160 to receive the blue side scattered light L12, and facilitates optical design of the first optical system 160. Further, since the blue side scattered light L12 can be received only by disposing the photodetector 134 on the negative X-axis side of the reflecting plate 173, the configuration of the entire optical system can be simplified.

  <Embodiment 2>

  As shown in FIG. 14, the second embodiment is different from the first embodiment in that the focal length of the second lens 171 is changed to f2, which is larger than f1. The diameter of the opening 172a of the second diaphragm member 172 is φ1 which is the same as the diameter of the opening 162a of the first diaphragm member 162. Other configurations of the second embodiment are the same as those of the first embodiment.

  As shown in FIG. 14, in the second embodiment, the focal length f2 is set to satisfy f2 / φ1 = f1 / φ2. Thereby, the numerical aperture of the second optical system 170 with respect to the flow cell 110 is the same as that of the first embodiment. Therefore, also in the second embodiment, the numerical aperture of the second optical system 170 with respect to the flow cell 110 is smaller than the numerical aperture of the first optical system 160 with respect to the flow cell 110. As a result, even if there is some variation in the arrangement of the optical components, the reflected light is unlikely to deviate from the light capturing range of the first optical system 160, so the same effect as in the first embodiment can be achieved. Note that the numerical aperture of the second optical system 170 for the flow cell 110 is not necessarily the same as that of the first embodiment, and the numerical aperture of the second optical system 170 for the flow cell 110 is the same as that of the first optical system 160 for the flow cell 110. It is sufficient if it is smaller than the numerical aperture.

  In the second embodiment, different lenses need to be used as the first lens 161 and the second lens 171, but the same member can be used as the first diaphragm member 162 and the second diaphragm member 172.

  Next, with reference to FIG. 15A to FIG. 18B, a simulation result of the second embodiment performed by the present inventor will be described. The simulation results shown below were performed under the same conditions as the simulation results shown in FIGS. 10 (a) to 13 (b). In addition, about a comparative example, since it is as showing to Fig.6 (a)-FIG.9 (b), description is abbreviate | omitted here.

  As shown in FIG. 15B, when the reflection-side member is not displaced, the reflected light traveling in the positive direction of the X axis is not shielded by the second diaphragm member 172 and the first diaphragm member 162. On the other hand, as shown in FIGS. 15A and 15B, when the reflection side member is shifted by 0.3 mm in the X-axis direction, the peripheral portion of the reflected light is the second diaphragm member 172 as in the first embodiment. However, the reflected light is not substantially shielded by the first diaphragm member 162.

  As shown in FIG. 16A, when the reflection-side member is displaced in the X-axis direction, the amount of received light detected by the photodetector 133 slightly decreases according to the amount of deviation. FIG. 16B is a graph obtained by normalizing the graph of FIG. 16A with the maximum amount of received light. As shown in FIG. 16B, the amount of received light when the amount of deviation in the X-axis direction is +0.3 mm is suppressed to a decrease of about 12% compared to the maximum amount of received light. Therefore, in the second embodiment, it can be seen that even when the reflection-side member is displaced in the X-axis direction, the rate of decrease in the amount of received light can be suppressed as compared with the comparative example.

  As shown in FIG. 17B, when the reflection side member is not displaced, the reflected light traveling in the positive X-axis direction is not shielded by the second diaphragm member 172 and the first diaphragm member 162. On the other hand, as shown in FIGS. 17A and 17C, when the reflection side member is shifted by 0.3 mm in the Y-axis direction, the peripheral portion of the reflected light is the second diaphragm member 172 as in the first embodiment. And is shielded from light by the first diaphragm member 162.

  As shown in FIG. 18A, when the reflection-side member is displaced in the Y-axis direction, the amount of received light detected by the photodetector 133 decreases according to the amount of deviation. FIG. 18B is a graph obtained by normalizing the graph of FIG. 18A with the maximum amount of received light. As shown in FIG. 18B, the amount of received light when the amount of deviation in the Y-axis direction is 0.3 mm is suppressed to a decrease of about 23% compared to the maximum amount of received light. Therefore, in Embodiment 2, it can be seen that even when the reflection-side member is displaced in the Y-axis direction, the rate of decrease in the amount of received light can be suppressed as compared with the comparative example.

  Note that the simulation results when the reflection-side member is displaced in the Z-axis direction are also the same as those shown in FIGS. 17 (a) to 18 (b). Therefore, it can be seen that even when the reflection-side member is displaced in the Z-axis direction, the decrease in the amount of received light can be suppressed lower than in the comparative example.

  <Embodiment 3>

  As shown in FIG. 19B, in the third embodiment, the focal length of the second lens 171 is changed to f3 smaller than f1, compared with the first embodiment (reposted in FIG. 19A). The diameter of the opening 172a of the aperture member 172 is changed to φ3 smaller than φ2. Other configurations of the third embodiment are the same as those of the first embodiment.

  As shown in FIG. 19B, the focal length f3 and the diameter φ3 are set to satisfy f3 / φ3 = f1 / φ2. Thereby, the numerical aperture of the second optical system 170 with respect to the flow cell 110 is the same as in the first and third embodiments. Therefore, also in the third embodiment, the numerical aperture of the second optical system 170 with respect to the flow cell 110 is smaller than the numerical aperture of the first optical system 160 with respect to the flow cell 110. As a result, even if there is some variation in the arrangement of the optical components, the reflected light is unlikely to deviate from the light capturing range of the first optical system 160, so the same effect as in the first embodiment can be achieved. Note that the numerical aperture of the second optical system 170 for the flow cell 110 is not necessarily the same as in the first and third embodiments, and the numerical aperture of the second optical system 170 for the flow cell 110 is the same as that of the first optical system for the flow cell 110. What is necessary is just to be smaller than the numerical aperture of the system 160.

  In the third embodiment, as shown in FIG. 19B, the diameter of the blue side scattered light L12 irradiated on the reflecting plate 173 is φ3, which is smaller than φ2 in the first embodiment. Thus, in the third embodiment, even if the light receiving surface 134a (see FIG. 1) of the photodetector 134 that receives the blue side scattered light L12 is reduced, the blue side scattered light L12 having the same light amount as that in the first embodiment is used. Can be received. For this reason, size reduction of the photodetector 134 can be achieved.

  Since the focal length f3 is smaller than the focal length f1, the flow diameter of the second diaphragm member 172 of the third embodiment is equal to φ2 as in the second diaphragm member 172 of the first embodiment. The numerical aperture of the second optical system 170 may be larger than the numerical aperture of the first optical system 160 for the flow cell 110 in some cases. However, in the third embodiment, since the aperture diameter φ3 of the second diaphragm member 172 is set smaller than the aperture diameter φ2 of the first embodiment, the numerical aperture of the second optical system 170 relative to the flow cell 110 is set to the flow cell 110. Is smaller than the numerical aperture of the first optical system 160.

  <Embodiment 4>

  As shown in FIG. 20, in the fourth embodiment, the configuration for irradiating and receiving the blue laser light L1 is omitted as compared with the first embodiment.

  Specifically, the light source 121, the light receiving surface 131 a of the photodetector 131, the photodetector 134, the collimator lens 141, the dichroic mirror 143, and the hole 153 a of the pinhole 153 are omitted, and the reflection plate 173. Instead, a mirror 174 that totally reflects light is added. The first lens 161 and the second lens 171 convert the red fluorescence L23 into parallel light. The dichroic mirror 163 reflects the red side scattered light L22 and transmits the red fluorescence L23. The spectral filter 165 transmits only the red fluorescence L23. Other configurations of the fourth embodiment are the same as those of the first embodiment.

  As shown in FIG. 20, the red side scattered light L22 and the red fluorescence L23 incident on the second optical system 170 are reflected by the mirror 174 and guided to the first optical system 160. The red side scattered light L22 incident on the first optical system 160 is received by the light receiving surface 132a of the photodetector 132, as in the first embodiment. The red fluorescence L23 incident on the first optical system 160 is received by the photodetector 133 as in the first embodiment. Also in this case, the reflected light reflected by the mirror 174 enters the first aperture member 162 at an angle θ2 smaller than the angle θ1, and a gap is generated between the reflected light and the opening 162a. Therefore, in the fourth embodiment, the same effect as in the first embodiment can be obtained.

  According to the fourth embodiment, the photodetector 133 receives red fluorescence L23 generated on the X-axis positive side and the X-axis negative side of the flow cell 110. Thereby, the detection light quantity of the red fluorescence L23 by the photodetector 133 can be increased. The photodetector 132 receives red side scattered light L22 generated on the X-axis positive side and the X-axis negative side of the flow cell 110. Thereby, the detection light quantity of the red side scattered light L22 by the photodetector 132 can be increased. Furthermore, the feature amount on the X axis positive side and the X axis negative side of the particle can be reflected in the red side scattered light L22 received by the photodetector 132.

  <Example of change>

  In the modification shown in FIG. 21A, the second lens 171 is omitted from the second optical system 170. In this case, the second optical system 170 includes a second diaphragm member 201 and a reflection plate 202. The second aperture member 201 includes a circular opening 201a having a diameter of φ2. The reflection plate 202 is a concave mirror, and includes a reflection surface 202 a that reflects the blue fluorescent light L <b> 13 that has passed through the second diaphragm member 201 and converges it on the flow path 115. The reflective surface 202a is disposed so that its optical axis is parallel to the X axis and penetrates the center of the flow path 115. The reflection surface 202a has, for example, a film structure that reflects only the blue fluorescence L13 and transmits other light. In this case, a configuration for guiding the blue side scattered light L12 is provided in the first optical system 160, and a photodetector for receiving the blue side scattered light L12 is provided in the optical detection unit 100.

  Also in the modification shown in FIG. 21A, the light included in the angle θ <b> 2 out of the light generated from the flow path 115 is returned to the first optical system 160 by the reflecting plate 202. Therefore, as in the first embodiment, the numerical aperture of the second optical system 170 with respect to the flow cell 110 is smaller than the numerical aperture of the first optical system 160 with respect to the flow cell 110.

  Further, instead of the first diaphragm member 162 and the second diaphragm member 172, other light limiting means may be provided.

  For example, as shown in FIG. 21B, a diffraction groove 211 may be provided on the incident surface of the second lens 171. In this case, the light incident on the diffraction groove 211 is diffracted outward, and only the light in the range of the angle θ2 is guided to the reflecting plate 173. Further, as illustrated in FIG. 21C, a light shielding film 221 may be provided on the incident surface of the second lens 171. In this case, the light incident on the light shielding film 221 is shielded, and only the light in the range of the angle θ2 is guided to the reflection plate 173. 21B and 21C, similarly to the first embodiment, the numerical aperture of the second optical system 170 with respect to the flow cell 110 is smaller than the numerical aperture of the first optical system 160 with respect to the flow cell 110. . The configuration of the first optical system 160 can also be changed in the same manner as in FIGS.

  Further, the light capturing range setting means is not limited to the diaphragm member. For example, in the first lens 161, the effective diameter that gives the lens action may be φ1. In this case, the first aperture member 162 is omitted. Further, in the second lens 171, the effective diameter for imparting the lens action may be φ2. In this case, the second aperture member 172 is omitted.

  In Embodiments 1 to 4, blood is the measurement target, but urine may be the measurement target. That is, the present invention is applicable to an apparatus for measuring particles in a biological sample such as blood or urine.

DESCRIPTION OF SYMBOLS 1 ... Particle measuring device 110 ... Flow cell 121 ... Light source 122 ... Light source 133 ... Photo detector 134 ... Photo detector 140 ... Optical system 160 ... First optical system 161 ... First lens 162 ... First aperture member 162a ... Aperture 170 ... Second optical system 171 ... Second lens 172 ... Second diaphragm member 172a ... Aperture 173 ... Reflector

Claims (11)

A flow cell for flowing particles;
A light source for irradiating the particles flowing in the flow cell with light;
A light detector for detecting light emitted from the particles flowing in the flow cell by being irradiated with light from the light source;
A first optical system disposed between the flow cell and the photodetector and guiding the light emitted from the particles to the photodetector;
A second optical system disposed on the side opposite to the first optical system with respect to the flow cell and reflecting light emitted from the particles to the first optical system,
The numerical aperture of the second optical system for the flow cell is smaller than the numerical aperture of the first optical system for the flow cell;
Particle measuring device. The particle measuring apparatus according to claim 1,
The first optical system includes a first lens on which light emitted from the particles flowing in the flow cell is incident upon irradiation with light from the light source,
The second optical system includes a second lens on which light emitted from the particles flowing in the flow cell is incident upon irradiation with light from the light source, and light transmitted through the second lens. A reflecting plate that reflects to the optical system 1,
The numerical aperture of the second lens is smaller than the numerical aperture of the first lens;
Particle measuring device. The particle measuring apparatus according to claim 2,
Each of the first lens and the second lens is a collimator lens.
Particle measuring device. In the particle measuring device according to claim 3,
The first optical system includes a first diaphragm member having an aperture diameter that defines a numerical aperture of the first lens,
The second optical system includes a second diaphragm member having an aperture diameter that defines the numerical aperture of the second lens,
The opening diameter of the second diaphragm member is smaller than the opening diameter of the first diaphragm member;
Particle measuring device. In the particle measuring device according to claim 4,
The focal length of the second lens is shorter than the focal length of the first lens;
Particle measuring device. In the particle measuring device according to claim 3,
The focal length of the second lens is longer than the focal length of the first lens;
Particle measuring device. In the particle measuring device according to any one of claims 2 to 6,
The reflecting plate transmits scattered light emitted from the particles when irradiated with light from the light source, and reflects fluorescence emitted from the particles when irradiated with light from the light source.
Particle measuring device. In the particle measuring device according to claim 7,
A second photodetector for receiving scattered light transmitted through the reflector;
Particle measuring device. In the particle measuring device according to any one of claims 2 to 8,
A first light source for irradiating the particles with light of a first wavelength;
A second light source for irradiating the particles with light having a second wavelength different from the first wavelength, as the light source,
Fluorescence generated from the particles when irradiated with light of the first wavelength, and scattered light and fluorescence generated from the particles when irradiated with light of the second wavelength are converted into the first optical system. A first light detection unit that receives light via
A second light detection unit that receives scattered light generated from the particles when irradiated with light of the first wavelength,
The reflecting plate emits only scattered light based on light of the first wavelength out of scattered light and fluorescence generated from the particles when irradiated with the light of the first wavelength and the light of the second wavelength. Transmits and guides to the second light detector, reflects at least fluorescence based on light of the first wavelength among the remaining light, and passes to the first light detector through the first optical system. Lead,
Particle measuring device. The particle measuring apparatus according to claim 9, wherein
From the first light source and the second light source, the light having the first wavelength and the light having the second wavelength are irradiated to the flow cell at positions shifted from each other in the flow direction of the particles. An optical system that guides the emitted light to the flow cell;
The reflecting plate absorbs fluorescence generated from the particles when irradiated with light of the second wavelength.
Particle measuring device. In the particle measuring device according to claim 9 or 10,
The reflector absorbs scattered light generated from the particles when irradiated with light of the second wavelength.
Particle measuring device.

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