JP2019035733A - Analyzer and analytical method - Google Patents

Abstract

To provide an analyzer and an analytical method for detecting and analyzing a materiality component in a sample at lower cost.SOLUTION: An analyzer includes: a flow cell 13A having a flow channel of a sample; a branch unit 102 for branching light transmitted through the flow channel into at least a first and a second optical paths; a first and a second imaging units 100A and 100B for shooting an image of the sample in the flow channel using light of the first and second optical paths; and a controller 14 for processing the shot image. The first and second imaging units 100A and 100B have a same view angle but shoot an image having different characteristics.SELECTED DRAWING: Figure 1

Description

  The present invention relates to an analysis apparatus and an analysis method.

  In the examination of the collected urine, it is considered to image the urine sample flowing through the flow cell channel formed of a transparent member through the wall of the flow cell. For example, by imaging a urine sample flowing through a flow path provided in the flow cell and analyzing the captured image, urinary sediment components (blood cells, epithelial cells, cylinders, bacteria, crystals, tangible ( A method of analyzing a solid component) is known (for example, see Patent Document 1). In the apparatus described in Patent Document 1, a sediment component in a urine sample passing through a detection region is detected by a detector, and after a delay time has elapsed, a pulse light source is emitted in an imaging region downstream from the detection region and a sediment component Is imaged.

Japanese Patent Laid-Open No. 06-288895

  However, since the flow rate of sediment components differs between the central portion of the flow cell flow path and near the wall surface, the passage position of sediment components in the flow cell is detected, and the delay time is adjusted based on this passage position. . For this reason, control becomes complicated. Further, an optical system for detecting the sediment component in the detection area and an optical system for imaging the sediment component in the imaging area are required, so that the apparatus becomes complicated and the manufacturing cost increases.

  An object of the present invention is to realize detection and analysis of a formed component in a specimen at a lower cost.

  One of the aspects of the present invention includes a flow cell having a flow path of a specimen including a formed component, a branching section that branches light transmitted through the flow path into at least a first optical path and a second optical path, and the first A first imaging unit and a second imaging unit configured to capture an image of the specimen in the flow channel using light of the optical path and light of the second optical path; and a control unit configured to process the captured image. The first imaging unit and the second imaging unit are analyzers that capture images having the same viewing angle but different characteristics.

  In addition, an aspect of the present invention includes a method invention corresponding to the analysis apparatus described above.

  According to the present invention, it is possible to realize detection and analysis of formed components in a specimen at a lower cost.

It is the figure which showed schematic structure of the analyzer which concerns on 1st Embodiment. It is the figure which showed schematic structure of the flow cell. It is the figure which showed schematic structure of the confluence | merging part and taper part vicinity. It is the figure which showed distribution of the sheath liquid which distribute | circulates a 4th channel | path, and a test substance. It is the flowchart which showed the flow which classifies the formed part. It is the figure which showed schematic structure of the analyzer which concerns on 2nd Embodiment. It is the flowchart which showed the flow which classifies the formed part. It is the figure which showed schematic structure of the analyzer which concerns on 3rd Embodiment.

  Embodiments for carrying out the present invention will be described below with reference to the drawings. However, the dimensions, materials, shapes, relative arrangements, and the like of the component parts described in this embodiment are not intended to limit the scope of the present invention only to those unless otherwise specified.

(First embodiment)
FIG. 1 is a diagram illustrating a schematic configuration of an analysis apparatus 20 according to the first embodiment. The analysis device 20 includes the imaging device 1. The imaging apparatus 1 analyzes, for example, a formed component in urine by imaging, for example, urine as a specimen and analyzing the captured image. However, the imaging device 1 can also be applied to the analysis of a formed component in a liquid sample other than urine such as blood or body fluid.

  The imaging apparatus 1 includes an imaging unit 10 that images a specimen, an imaging light source 12, and a flow cell unit 13. The flow cell unit 13 includes a stage (not shown) on which a flow cell 13A through which a sample flows is fixedly arranged. The flow cell 13A may be detachable from the stage.

The imaging apparatus 1 includes an objective lens 101, a branching unit 102, a first lens group 103A, a second lens group 103B, a mask 104, a first camera 105A, and a second camera 105B. The first camera 105A and the second camera 105B are, for example, CCD (Charge Coupled Device).
Imaging is performed using an image sensor such as an image sensor or a complementary metal oxide semiconductor (CMOS) image sensor. Hereinafter, the objective lens 101, the branching unit 102, the first lens group 103A, the mask 104, and the first camera 105A are collectively referred to as a first imaging unit 100A, and the objective lens 101, the branching unit 102, the second lens group 103B, The two cameras 105B are collectively referred to as a second imaging unit 100B. Each of the first lens group 103A and the second lens group 103B includes an eyepiece lens, and may further include an imaging lens. The flow cell 13A is disposed between the light source 12 and the objective lens 101, and the light source 12 and the objective lens 101 are shared by the first imaging unit 100A and the second imaging unit 100B.

  The analyzer 20 is provided with a controller 14 as a control unit. The controller 14 includes a CPU 14A, a ROM 14B, a RAM 14C, an EEPROM 14D, and an interface circuit 14E, and is connected to each other by a bus line 14F.

A central processing unit (CPU) 14A operates based on a program stored in a read only memory (ROM) 14B and read into a random access memory (RAM) 14C, and controls the entire analyzer 20. The ROM 14B stores programs and data for operating the CPU 14A. The RAM 14C provides a work area to the CPU 14A and temporarily stores various data and programs. An EEPROM (electrically erasable programmable read only memory) 14D stores various setting data. The interface circuit 14E controls communication between the CPU 14A and various circuits.

The interface circuit 14E is connected to control lines of the first imaging unit 100A, the second imaging unit 100B, the light source 12, the first pump 15A, and the second pump 15B, and these devices receive control signals from the controller 14. Controlled by. The first pump 15A is a pump that supplies the sheath liquid to the flow cell 13A via the first supply pipe 132A, and the second pump 15B is a pump that supplies the specimen to the flow cell 13A via the second supply pipe 133A. .
The sheath liquid is a liquid that controls the flow of the specimen in the flow cell 13A. For example, when the specimen is urine, physiological saline is applied. However, a solution other than physiological saline may be used as the sheath liquid.

  The branching unit 102 branches light from the flow cell 13A in two or more directions. The branching unit 102 is, for example, a splitter such as a half mirror, and splits light in two directions by transmitting part of the light that has passed through the objective lens 101 and reflecting the rest. Of the branched light, the light transmitted through the branching portion 102 is incident on the first lens group 103A and is incident on the imaging surface of the image sensor included in the first camera 105A. That is, it is used for imaging in the first imaging unit 100A. On the other hand, the light reflected by the branching unit 102 enters the second lens group 103B and enters the imaging surface of the imaging element of the second camera 105B. That is, the second imaging unit 100B is used for imaging. An optical path of light between the branching unit 102 and the first camera 105A is referred to as a first optical path, and an optical path of light between the branching unit 102 and the second camera 105B is referred to as a second optical path. Further, as shown in FIG. 1, the branching portion 102 is disposed on the optical axis 11 </ b> B of the objective lens 101. In FIG. 1, the optical axis of the first optical path is indicated by 111A, and the optical axis of the second optical path is indicated by 111B.

  The mask 104 is disposed between the branch portion 102 and the first lens group 103A (that is, inserted into the first optical path). The mask 104 is formed by making a circular hole in the plate, and the optical axis 111A of the first lens group 103A passes through the central axis of the hole of the mask 104 and is arranged at a position orthogonal to the optical axis 111A of the first optical path. Yes. The mask 104 functions as an aperture that reduces the amount of light traveling toward the first camera 105A by blocking part of the light in the first optical path. The mask 104 increases the depth of field of the first camera 105A.

  FIG. 2 is a diagram showing a schematic configuration of the flow cell 13A. The flow cell 13A is formed by joining (for example, thermocompression bonding) the first plate 130 and the second plate 131. FIG. 2 is a view of the flow cell 13A viewed from the first plate 130 side. In addition, let the width direction of the flow cell 13A shown in FIG. 2 be the X-axis direction in the orthogonal coordinate system, the length direction be the Y-axis direction, and the thickness direction be the Z-axis direction. The sample to be imaged flows in the Y-axis direction in the flow cell 13A. The optical axis 11B of the objective lens 101 is arranged in the Z-axis direction.

  For the material of the flow cell 13A, a material having visible light transmissivity of 90% or more such as PMMA (acrylic resin), COP (cycloolefin polymer), PDMS (polydimethylsiloxane), PP (polypropylene), and quartz glass is adopted. be able to.

  The first plate 130 is provided with a first supply port 132 for supplying a sheath liquid, a second supply port 133 for supplying a specimen, and a discharge port 134 for discharging the sheath liquid and the specimen. The first supply port 132, the second supply port 133, and the discharge port 134 pass through the first plate 130, respectively. The first supply port 132 is provided on one end side in the longitudinal direction of the first plate 130, the second supply port 133 is provided on the other end side in the longitudinal direction of the first plate 130, and the discharge port 134 is provided on the first plate 130. The plate 130 is provided between the first supply port 132 and the second supply port 133 in the longitudinal direction.

  The 1st supply port 132, the 2nd supply port 133, and the discharge port 134 are mutually connected by channel | path 135A, 135B, 136,138. These passages 135 </ b> A, 135 </ b> B, 136, and 138 are formed so as to be recessed from the surface on the joining surface side of the first plate 130 so that the cross section is rectangular. Further, the sections of the passages 135A, 135B, 136, and 138 are formed so that the width direction (X-axis direction in FIG. 2) is larger than the depth direction (Z-axis direction in FIG. 2). When the first plate 130 and the second plate 131 are joined, the second plate 131 becomes a wall material that forms the passages 135A, 135B, 136, and 138.

  A first passage 135 </ b> A and a second passage 135 </ b> B are connected to the first supply port 132. The first passage 135 </ b> A and the second passage 135 </ b> B are reversely rotated in the reverse direction along the outer edge of the first plate 130 toward the second supply port 133, and merge at the junction 137. A third passage 136 is connected to the second supply port 133, and the third passage 136 joins the first passage 135A and the second passage 135B at the joining portion 137. The junction 137 is connected to the outlet 134 via the fourth passage 138. The fourth passage 138 has a tapered shape in which the depth of the fourth passage 138 (the length in the thickness direction (Z-axis direction) of the first plate 130) gradually decreases from the junction 137 toward the discharge port 134. The formed taper portion 138A is formed. The taper portion 138A is provided with an inclination of 2 ° to 8 °, for example.

  1 is connected to the first supply port 132, the second supply tube 133 </ b> A shown in FIG. 1 is connected to the second supply port 133, and the discharge port 134 is connected to the first supply port 132. A discharge pipe (not shown) is connected. The sheath liquid supplied from the first supply pipe 132A to the first supply port 132 flows through the first passage 135A and the second passage 135B. The specimen supplied from the second supply pipe 133A to the second supply port 133 flows through the third passage 136. Then, the sheath liquid and the sample are joined at the joining portion 137, flow through the fourth passage 138, and are discharged from the discharge port 134 to the discharge pipe.

  FIG. 3 is a diagram showing a schematic configuration in the vicinity of the merging portion 137 and the tapered portion 138A. In the merging portion 137, the third passage 136 is arranged to be biased toward the second plate 131, and the specimen flows along the second plate 131 in the merging portion 137.

FIG. 4 is a view showing the distribution of the sheath liquid and the sample flowing through the fourth passage 138. After the sheath liquid and the sample are separately supplied from the upper side in FIG. Immediately after the sheath liquid and the specimen merge at the junction 137, the specimen in the sheath liquid is concentrated in a relatively narrow range on the wall surface side of the second plate 131 (see the section AA in FIG. 4). Thereafter, when the specimen flows through the tapered portion 138A, the specimen is pushed by the sheath liquid and spreads in a flat shape along the wall surface near the wall surface of the second plate 131 (see the section BB in FIG. 4). When the sample further flows, the sample is separated from the wall surface of the second plate 131 by the Tubular-pinch effect, and is lifted toward the center of the fourth passage 138 (refer to the CC cross section in FIG. 4).

  The distribution of the formed component is affected by the distribution of the specimen fluid in the sheath liquid. By performing imaging at a position where a larger amount of the formed portion can be imaged, the analysis accuracy of the formed portion can be increased. In the flow cell 13A, as shown in the sectional view of FIG. 4, the flow of the specimen changes depending on the position in the Y-axis direction. In the position of the CC cross section in FIG. 4, the width of the specimen in the Z-axis direction is larger than the position of the BB cross section. At the position of the CC cross section in FIG. 4, the formed component in the specimen spreads in the Z-axis direction and is not suitable for imaging the formed component.

On the other hand, at the position of the BB cross section of FIG. 4, the sheath liquid flows from above to press the specimen against the second plate 131, and the specimen is crushed by the sheath liquid and spreads thinly. Therefore, at the position of the BB cross section in FIG. 4, the formed portion in the specimen exists without spreading in the Z-axis direction, and is easily focused. In addition, the sheath fluid and the specimen fluid form a laminar flow and hardly mix. Since such a position of the BB cross section is a position in the Y-axis direction suitable for imaging the formed portion, the specimen is imaged at the position in the Y-axis direction. This position is called an imaging position, and the optical axis 11B of the objective lens 101 is aligned with this imaging position.

In the first imaging unit 100A, the mask 104 is inserted into the first optical path to reduce the amount of light toward the first camera 105A (the numerical aperture is reduced). On the other hand, the aperture corresponding to the mask 104 is not provided in the second optical path. For this reason, the depth of field of the image captured by the first imaging unit 100A is deeper than the depth of field of the image captured by the second imaging unit 100B. On the other hand, the resolution of the second imaging unit 100B is higher than the resolution of the first imaging unit 100A. Note that the diameter of the hole formed in the mask 104 is set to have a desired numerical aperture in the first imaging unit 100A. The desired numerical aperture can be set according to the depth of field required for the first imaging unit 100A.

  Further, the depth of field of the second imaging unit 100B is included so that the depth of field of the image captured by the second imaging unit 100B is included in the depth of field of the image captured by the first imaging unit 100A. Is set.

  The first camera 105 </ b> A and the second camera 105 </ b> B simultaneously capture the formed still image in the sample flowing through the flow cell 13 </ b> A. The imaging is enlarged imaging, and the lighting time of the light source 12 and the imaging time (exposure time) of the first camera 105 </ b> A and the second camera 105 </ b> B are synchronized by the controller 14. Parallel light enters the flow cell 13A from the light source 12. During imaging, the light source 12 is turned on one or more times. The lighting time of the light source 12 depends on the flow rate of the specimen, and is set to 0.1 to 10 μsec, for example, so that the subject shake is within the allowable range. The number of formed portions included in one image may be increased by causing the light source 12 to emit light a plurality of times for one exposure. By imaging more of the formed component, the analysis accuracy of the formed component can be further increased. The blinking timing of the light source 12 in this case is determined in consideration of the relationship between the flow rate of the specimen and the lighting time of the light source 12 so that the same specimen is not imaged. For example, a xenon lamp or a white LED can be used as the light source 12, but the present invention is not limited to this, and other light sources can also be used. A plurality of such images are taken.

  The optical path is branched into the first optical path and the second optical path by the branching unit 102, and the mask 104 is provided only in the first optical path out of the first optical path and the second optical path, so that the light from the light source 12 that has passed through the flow cell 13A. When images of light are simultaneously captured by the first imaging unit 100A and the second imaging unit 100B, two images having the same viewing angle and different characteristics such as depth of field and resolution can be acquired. Since the image captured by the first imaging unit 100A having a depth of field deeper than the second imaging unit 100B has a wide range in focus on the formed portion, it is suitable for obtaining the position and number of the formed portion. On the other hand, the image picked up by the second image pickup unit 100B having a higher resolution is suitable for classifying the formed components classified by type observation and type observation of cell nuclei and the like in more detail.

  The CPU 14A performs imaging of the specimen flowing through the flow cell 13A using the first imaging unit 100A and the second imaging unit 100B, and determines the position, size, and number of formed components from the image captured by the first imaging unit 100A. It grasps | ascertains, determines the cutout size of an image from the magnitude | size of the grasped formation, and produces | generates a cutout image. The cut-out image is an image obtained by comparing the background image and the captured image, and cutting out the image inside the portion surrounded by a square.

  Prior to the generation of the cut-out image, the CPU 14A creates, as a background image, an average of the pixel values of each pixel for each image using the stored image data. The pixel value may be the luminance of each pixel or an RGB value. The cutout image is generated when the CPU 14A executes a program (cutout process) stored in the ROM 14B. The cutout image is stored in the RAM 14C together with the cutout position and cutout size. For example, the CPU 14A generates cut-out images for all formed components included in the captured image.

The CPU 14A captures an image having the same cutout position and the same cutout size as the cutout image generated from the image picked up by the first image pickup unit 100A (hereinafter referred to as the first cutout image) by the second image pickup unit 100B. To generate a cutout image (hereinafter referred to as a second cutout image). Then, the first cutout image and the second cutout image are associated with each other and stored in the RAM 14C. The first cutout image and the second cutout image are CPU
14A is used for various analyses.

  Since the formed component can be recognized from the image captured by the first imaging unit 100A, it is not necessary to perform a process of recognizing the formed component from the image captured by the second image capturing unit 100B. Thus, based on the image imaged by the first imaging unit 100A, the position, size, and number of the formed components are grasped, and the formed image is obtained based on the image captured by the second imaging unit 100B. Conduct minute morphological observations. This morphological observation can analyze and classify the formed component with high accuracy.

  FIG. 5 is a flowchart showing a flow of classifying the formed component. This flowchart is executed by the CPU 14A.

  In step S101, the CPU 14A acquires images captured by the first imaging unit 100A and the second imaging unit 100B. Hereinafter, an image captured by the first imaging unit 100A is referred to as a first image, and an image captured by the second imaging unit 100B is referred to as a second image.

  When the process of step S101 is completed, the process proceeds to step S102, and the CPU 14A generates a first cutout image by cutting out the formed portion from the first image, and stores it in the RAM 14C.

  When the process of step S102 is completed, the process proceeds to step S103, and the CPU 14A acquires the position information and the feature amount of the first clipped image. The position information and the feature amount of the first cutout image are stored in the RAM 14C in association with the first cutout image. Examples of the feature amount include color, shape, and size. A program stored in advance in the ROM 14B is used to acquire the feature amount.

  When the process of step S103 is completed, the process proceeds to step S104, and the CPU 14A cuts out the formed portion from the second image and generates a cutout image (second cutout image). The second cutout image is generated by cutting out an image having the same size at the same position as the first cutout image based on the position information of each of the plurality of first cutout images.

  When the process of step S104 is completed, the process proceeds to step S105, and the CPU 14A acquires the feature amount of the second clipped image. The feature amount of the second clipped image is stored in the RAM 14C in association with the second clipped image.

  When the process of step S105 is completed, the process proceeds to step S106, and the CPU 14A classifies the formed portion based on the feature amounts acquired in step S103 and step S105. For the classification, a program stored in the ROM 14B in advance is used. For example, the CPU 14A classifies the formed portion by comparing the feature amount of the first cutout image or the feature amount of the second cutout image with the feature amount of each formed portion stored in the ROM 14B in advance.

  When the process of step S106 is completed, the process proceeds to step S107, and the CPU 14A counts the formed component for each type of formed component classified in step S106, and then proceeds to step S108 to output the count result in step S107. . Based on the counting result, the CPU 14A may perform various analyses.

In this way, the position and number of the formed portion can be specified based on the first cutout image, and the analysis accuracy of the formed portion can be improved by classifying the formed portion based on the second cutout image. Can be increased. Further, when acquiring images having different characteristics such as the first image and the second image, only one light source 12 and one objective lens 101 are required. In addition, by simultaneously capturing the first image and the second image having the same viewing angle, it is not necessary to provide a delay time for capturing the first image and the second image, thereby simplifying the device and the control. Can do. Thereby, detection and analysis of the formed component in the specimen can be realized at a lower cost.

  In the above description, the mask 104 is used as an optical member that reduces the amount of light, but an optical slit may be used instead.

(Second Embodiment)
In the first embodiment, the depth of field and resolution of the first image are changed by the mask 104. In the second embodiment, instead of arranging the mask 104 in the first imaging unit 100A, The optical filter 106 is disposed in the second imaging unit 100B. In the following description, differences from the first embodiment will be mainly described.

  FIG. 6 is a diagram illustrating a schematic configuration of the analyzer 20 according to the second embodiment. In the first imaging unit 100A, the mask 104 is not provided. In the second imaging unit 100B, an optical filter 106 is provided between the second camera 105B and the second lens group 103B. The optical filter 106 is, for example, an optical filter that transmits only light having a predetermined wavelength.

  For example, a case where red blood cells are detected as a sediment component in a urine sample will be described. Since erythrocytes absorb green, an optical filter 106 that transmits green can be provided, and the formed component other than erythrocytes in the sample can be imaged by the second camera 105B. At this time, the formed portion including red blood cells is captured in the image captured by the first camera 105A at the same time. When the formed component that can be recognized in the first image cannot be recognized in the second image, the formed component can be classified as red blood cells. Therefore, it is possible to accurately distinguish erythrocytes from calcium oxalate crystals that resemble erythrocytes. The transmission wavelength of the optical filter 106 can be appropriately selected according to the specimen fluid and the formed component. The optical filter 106 can be easily replaced according to the component to be distinguished.

  As described above, by using the optical filter 106, it is possible to easily classify the formed portion in the specimen. The CPU 14A generates a first cutout image from the first image, generates a second cutout image having the same position and size as the first cutout image from the second image, compares the generated image with the first cutout image, and classifies the formed portion. Do.

  FIG. 7 is a flowchart showing a flow for classifying the formed component. This flowchart is executed by the CPU 14A.

  From Step S201 to Step S205, the CPU 14A executes the same processing as Step S101 to Step S105 shown in FIG.

  When the process of step S205 is completed, the process proceeds to step S206, and the CPU 14A classifies the formed portion based on the feature amounts acquired in step S203 and step S205. For example, if the first cut-out image can recognize the formed component but cannot recognize the formed component in the corresponding second cut-out image at the same position, the corresponding filter corresponds to light having a wavelength that could not be transmitted by the optical filter 106. Classified as forming. For the classification, a program stored in the ROM 14B in advance is used.

When the process of step S206 is completed, the process proceeds to step S207, and the CPU 14A counts the formed components classified in step S206, and then proceeds to step S208 to output the count result of step S207. Based on the counting result, the CPU 14A may perform various analyses. Further, the CPU 14A may analyze the formed component based on the classified first cutout image, or may analyze the formed component based on an image in which the formed component can be recognized in the second cutout image. .

  As described above, by classifying the formed portion based on the first cut-out image and the second cut-out image, the analysis accuracy of the formed portion can be increased. Further, by inserting the optical filter 106 in one of the plurality of optical paths, the wavelength characteristics of light incident on the first imaging unit 100A and the second imaging unit 100B are different. Therefore, when acquiring images having different characteristics such as the first image and the second image, only one light source 12 and one objective lens 101 are required. In addition, by photographing the same field of view at the same time, it is not necessary to provide a delay time for imaging the first image and the second image, so that the apparatus and control can be simplified. Thereby, detection and analysis of the formed component in the specimen can be realized at a lower cost.

(Third embodiment)
In the first embodiment and the second embodiment, the first imaging unit 100A and the second imaging unit 100B are provided as imaging units, but more imaging units can be provided. The mask 104 according to the first embodiment and the optical filter 106 according to the second embodiment can be combined. In addition, a plurality of masks having different hole diameters can be provided in different imaging units. In addition, a plurality of optical filters having different wavelengths of transmitted light can be provided in different imaging units. Moreover, a slit can also be provided. Hereinafter, a configuration in which three imaging units are provided will be described.

  FIG. 8 is a diagram illustrating a schematic configuration of the analyzer 20 according to the third embodiment. The imaging device 1 includes a first imaging unit 100A, a second imaging unit 100B, and a third imaging unit 100C. In addition, the imaging device 1 includes a first branch portion 102A and a second branch portion 102B. The first imaging unit 100A includes an objective lens 101, a first branching unit 102A, a first lens group 103A, a mask 104, and a first camera 105A. The second imaging unit 100B includes an objective lens 101, a first branching unit 102A, a second branching unit 102B, a second lens group 103B, an optical filter 106, and a second camera 105B. The third imaging unit 100C includes an objective lens 101, a first branching unit 102A, a second branching unit 102B, a third lens group 103C, and a third camera 105C.

  The first lens group 103A, the second lens group 103B, and the third lens group 103C each include an eyepiece, and may further include an imaging lens. The objective lens 101 and the light source 12 are shared by the first imaging unit 100A, the second imaging unit 100B, and the third imaging unit 100C.

  The first branching part 102A and the second branching part 102B are splitters such as a half mirror. The first branching part 102A splits light in two directions by transmitting part of the light that has passed through the objective lens 101 and reflecting the rest. Light that passes through the first branch portion 102A is incident on the first lens group 103A, and is used for imaging in the first imaging portion 100A.

  The second branch portion 102B further splits the light in two directions by transmitting part of the light reflected by the first branch portion 102A and reflecting the rest. Light that passes through the second branch portion 102B is incident on the second lens group 103B, and is used for imaging in the second imaging portion 100B. The light reflected by the second branching section 102B is incident on the third lens group 103C, and is used for imaging by the third imaging section 100C.

  The controller 14 is connected to the first imaging unit 100A, the second imaging unit 100B, the third imaging unit 100C, the light source 12, and the first pump 15A and the second pump 15B. Controlled by.

  The CPU 14A simultaneously captures images with the same viewing angle using the first imaging unit 100A, the second imaging unit 100B, and the third imaging unit 100C. Then, from the image captured by the first imaging unit 100A, the position, size, and number of formed components are grasped, and the formed components in the specimen are captured from the images captured by the second imaging unit 100B and the third imaging unit 100C. Minute classification is performed, and fine morphological observation of the formed portion is performed based on the image captured by the third imaging unit 100C. The hole diameter of the mask 104 and the type of the optical filter 106 are appropriately selected according to the type of formed component and the analysis content. In the analysis of the formed component, the same processing as the flowchart shown in FIG. 5 or FIG. 7 may be performed.

  As described above, even when three or more imaging units are provided, it is possible to simultaneously capture images with the same viewing angle, so that the apparatus and control can be simplified. Thereby, detection and analysis of the formed component in the specimen can be realized at a lower cost.

  In the above embodiment, for example, an optical member that reduces the amount of incident light on the first camera 105A is provided in the first imaging unit 100A, or an optical filter 106 is provided in the second imaging unit 100B. Needless to say, an optical member or an optical filter may be provided in any one imaging unit.

  In the above-described embodiment, the sample after passing through the tapered portion 138A of the flow cell 13A has been described as an example in which the sample is in contact with the wall surface of the flow cell 13A. However, the flow cell structure and the sample flow are limited to this mode. Not. For example, a flow cell having a structure in which the sheath liquid surrounds the specimen after passing through the tapered portion 138A of the flow cell 13A and the specimen is thinly stretched at the center of the sheath liquid may be used. Needless to say, the first camera 105A and the second camera 105B used in the above embodiment are not particularly limited, and the same camera may be used or cameras having different performances may be used.

(Fourth embodiment)
This embodiment will be described with reference to FIG. In the present embodiment, the number of pixels of the imaging element included in the first camera 105A of the first imaging unit 100A (hereinafter also referred to as the number of pixels of the first camera 105A or the number of pixels of the first imaging unit 100A) is the first. The number of pixels of the image sensor of the second camera 105B of the second imaging unit 100B (hereinafter also referred to as the number of pixels of the second camera 105B or the number of pixels of the second imaging unit 100B) is smaller. Since the first imaging unit 100A includes the mask 104, the depth of field of the image captured by the first imaging unit 100A is greater than the depth of field of the image captured by the second imaging unit 100B. Is also deeper. On the other hand, the resolution of the second imaging unit 100B is higher than the resolution of the first imaging unit 100A. The viewing angle of the first imaging unit 100A and the viewing angle of the second imaging unit 100B are the same.

  Here, since there is an appropriate number of pixels of the camera with respect to the resolution of the imaging unit, when a camera having a number of pixels that does not consider the resolution of each imaging unit is used, the number of pixels is small with respect to the resolution. It may be (less) too much or too large (more). When the number of pixels is too large (more) than the resolution, it means that the size of one pixel is too small for the resolution. That the number of pixels is too small (small) with respect to the resolution means a state where the size of one pixel is too large with respect to the resolution. If a camera having the number of pixels corresponding to the resolution of the second imaging unit 100B is used for the first imaging unit 100A, the first imaging unit 100A may have an unnecessarily large number of pixels with respect to the resolution. On the other hand, when a camera having the number of pixels corresponding to the resolution of the first imaging unit 100A is used for the second imaging unit 100B, the number of pixels in the second imaging unit 100B may be too small with respect to the resolution.

On the other hand, by making the number of pixels of the first camera 105A smaller than the number of pixels of the second camera 105B, a camera having the number of pixels corresponding to the resolution of each of the first imaging unit 100A and the second imaging unit 100B can be obtained. Can be arranged. The combinations of the number of pixels of the first camera 105A and the second camera 105B include, for example, (1) the number of pixels of the first camera 105A is 2 million to 3 million pixels, and the number of pixels of the second camera 105B is 1800. (2) A combination in which the number of pixels of the first camera 105A is 2 million to 3 million pixels and a number of pixels of the second camera 105B is 5 million to 12 million pixels, (3) A combination in which the number of pixels of one camera 105A is 1 million pixels or less and the number of pixels of the second camera 105B is 2 million to 3 million pixels is conceivable, but the combination of the number of pixels is not limited thereto. For example, the number of pixels of the first camera 105 </ b> A may be any number of pixels suitable for obtaining the position and number of formation. On the other hand, the number of pixels of the second camera 105B only needs to be the number of pixels suitable for observation of fine forms for formation. By selecting such a number of pixels, the number of pixels of the first camera 105A becomes smaller than the number of pixels of the second camera 105B.

  In general, the smaller the number of pixels of the camera, the lower the price of the camera. Therefore, the price of the first camera 105A can be reduced by making the number of pixels of the first camera 105A smaller than the number of pixels of the second camera 105B. Thus, the cost of the entire analyzer 20 can be reduced. In general, the smaller the number of pixels, the smaller the load of image processing. Therefore, by making the number of pixels of the first camera 105A smaller than the number of pixels of the second camera 105B, processing of images captured by the first camera 105A is performed. The speed can be increased, and thus the processing speed of the analyzer 20 as a whole can be increased.

DESCRIPTION OF SYMBOLS 1 Imaging device 12 Light source 13A Flow cell 14 Controller 20 Analyzer 100A 1st imaging part 100B Second imaging part 101 Objective lens 102 Branch part 104 Mask

Claims (8)

A flow cell having a flow path for a specimen containing a formed component;
A branching portion for branching the light transmitted through the flow path into at least a first optical path and a second optical path;
Using the light of the first optical path and the light of the second optical path, a first imaging unit and a second imaging unit that capture an image of the specimen in the channel;
A control unit for processing the captured image;
With
The first imaging unit and the second imaging unit are analyzers that capture images having the same viewing angle but different characteristics.   The control unit generates position information for the formed component from the image captured by the first image capturing unit, and the image for the formed component from the image captured by the second image capturing unit based on the position information. The analysis apparatus according to claim 1, wherein a process of cutting out the first is performed.   The controller is configured to capture the second image using an image captured by the first imaging unit using light branched from the branch unit to the first optical path and light branched from the branch unit to the second optical path. The analyzer according to claim 1 or 2, wherein the detection processing for the formed component is performed in comparison with an image captured by the unit.   The first imaging unit includes an optical member that reduces the amount of incident light, and the depth of field of the first imaging unit is larger than the depth of field of the second imaging unit. 2. The analyzer according to item 1.   5. The second imaging unit includes an optical filter that transmits light of a predetermined wavelength, and the wavelength characteristics of light incident on the first imaging unit and light incident on the second imaging unit are different from each other. The analyzer according to any one of the above.   The analyzer according to claim 1, wherein the number of pixels of the first imaging unit is smaller than the number of pixels of the second imaging unit. Irradiating light from a light source to a flow cell having a flow path of a specimen including a formed component;
Branching light transmitted through the flow path into at least a first optical path and a second optical path;
Capturing a first image that is an image of the specimen in the flow path using light of the first optical path;
Capturing a second image having the same viewing angle as the first image using light in the second optical path, but having different characteristics;
Cutting out a first cutout image including the formed portion from the first image, and specifying position information of the first cutout image;
Cutting out the second cut-out image of the formed portion from the second image based on the position information;
Analytical methods including:   The analysis method according to claim 7, further comprising a step of detecting the formed portion by comparing the first cutout image and the second cutout image.

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