Optical systems and specimen analyzers

Abstract

To provide an optical system and a specimen analyzer capable of achieving miniaturization.SOLUTION: An optical system 100 includes: a light source 111 for emitting light; an irradiation optical system IS for irradiating illumination light in which a plurality of diffracted light beams generated by a diffraction optical element 114 are distributed; a flow cell 101 in which a sample containing cells is caused to flow; and light receiving units 123, 133, 143 for receiving light generated from the cells flowing in the flow cell 101 by irradiation of the illumination light. The system irradiates illumination light including zero-order diffracted light whose relative intensity relative to another diffracted light is 10 times or less at a position where the cells in the flow cell 101 pass.SELECTED DRAWING: Figure 3

Description

Technical Field

[0004] , , , , , ,

[0001] The present invention relates to an optical system and a specimen analyzer.

Background Art

[0002] As a method for accurately and quickly classifying cells contained in a sample, the following Patent Document 1 describes a method called ghost cytometry (registered trademark). In this method, patterned structured light illumination is applied to a sample containing cells, and the classification of the cells is performed by inputting the information of the light generated from the cells into a machine learning classifier. For example, like the optical system shown in FIG. 15, the structured light illumination is generated using diffraction optical elements DOE-1 and DOE-2, and is applied to a sample containing cells through an objective lens 1004 and a fluorescent film 1001. The structured illumination generated by the diffraction optical elements DOE-1 and DOE-2 is illuminated on a combined image plane using a lens 1002 having a focal length of 150 mm in order to suppress the influence of noise on the cell-derived signals required for analysis, and a diffraction pattern including zero-order and multiple-order diffracted lights is blocked by a spatial filter arranged on the combined image plane. The structured illumination with the diffraction pattern blocked is relayed by a lens 1003 having a focal length of 150 mm and is applied to the sample through the objective lens 1004 and the fluorescent film 1001.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0004] In the configuration described in Patent Document 1, a configuration is provided to block the diffraction pattern in order to suppress the influence of noise on the cell-derived signal necessary for analysis, which leads to the problem of the optical system becoming large. Specifically, in the configuration described in Patent Document 1, two lenses 1002 and 1003 are required to position the spatial filter that blocks the diffraction pattern. The two lenses 1002 and 1003 are placed 300 mm apart, taking into account the focal length, which is a factor in the large size of the optical system.

[0005] In view of these challenges, the present invention aims to provide an optical system and a sample analyzer that enable miniaturization while suppressing the influence of noise on cell-derived signals necessary for analysis. [Means for solving the problem]

[0006] The optical system (100) of the present invention includes a light source (111) that emits light and a diffractive optical element (114) to which light is incident, and a plurality of diffracted light generated by the diffractive optical element (114) spot but In the irradiation area (R), in a predetermined pattern An illumination optical system (IS) that emits distributed illumination light, Irradiation area (R) A flow cell (101) through which a sample containing cells flows, and illumination light from an illumination optical system (IS) is used to emit light from the cells flowing through the flow cell (101). Without separating by diffracted light The system comprises light-receiving sections (123, 133, 143), and the illumination light includes zero-order diffracted light whose relative intensity to other diffracted light is 10 times or less, and the illumination optical system (IS) is Multiple diffracted light spots in the irradiation area (R) 0th-order diffracted light spot Includes Meru .

[0007] This optical system eliminates the need for a configuration to block zero-order diffracted light, thus allowing for a compact optical system while minimizing the impact of noise on cell-derived signals necessary for analysis.

[0008] The specimen analyzer (1) of the present invention includes a specimen preparation unit (21) that mixes a specimen containing cells with a reagent to prepare a specimen, a light source (111) that emits light, and a diffractive optical element (114) to which light is incident, wherein multiple diffracted light generated by the diffractive optical element (114) spot but In the irradiation area (R), in a predetermined pattern Distributed illumination light statically The irradiation optical system (IS) that provides the illumination, Irradiation area (R) A flow cell (101) through which a sample containing cells flows, and illumination light from an illumination optical system (IS) is used to emit light from the cells flowing through the flow cell (101). Without separating by diffracted light The light-receiving units (123, 133, 143) and the signals from the light-receiving units (122, 133, 143) AI algorithm The system comprises a control unit (10) for classifying cells, and the illumination light includes zero-order diffracted light whose relative intensity to multiple other diffracted lights is 10 times or less, and the illumination optical system (IS) is Multiple diffracted light spots in the irradiation area (R) 0th-order diffracted light spot Includes Meru .

[0009] This sample analyzer eliminates the need for a configuration to block zero-order diffracted light, thus enabling miniaturization of the sample analyzer while suppressing the impact of noise on cell-derived signals necessary for analysis. [Effects of the Invention]

[0010] According to the present invention, it is possible to miniaturize optical systems and sample analyzers. [Brief explanation of the drawing]

[0011] [Figure 1] Figure 1 is a block diagram showing the configuration of a sample analyzer according to Embodiment 1. [Figure 2] Figure 2 is a schematic diagram showing the AI ​​algorithm before and after training according to Embodiment 1. [Figure 3] Figure 3 is a schematic diagram showing the configuration of the optical system according to Embodiment 1. [Figure 4] Figure 4 is a schematic diagram showing the flow cell and illumination light according to Embodiment 1. [Figure 5] FIG. 5 is a diagram schematically showing a distribution pattern of diffracted light included in illumination light according to Embodiment 1. [Figure 6] FIG. 6 is a diagram showing detection signals obtained based on lymphocytes in an experiment according to Embodiment 1. [Figure 7] FIG. 7 is a diagram showing detection signals obtained based on neutrophils in an experiment according to Embodiment 1. [Figure 8] FIG. 8 is a diagram showing experimental results of classification performance when the length of illumination light is changed according to Embodiment 1. [Figure 9] FIG. 9 is a diagram showing simulation results of detection signals corresponding to relative intensities according to a comparative example and Embodiment 1. [Figure 10] FIG. 10 is a diagram showing simulation results of detection signals corresponding to relative intensities according to Embodiment 1. [Figure 11] FIG. 11 is a diagram showing simulation results of the performance of a diffractive optical element when the spot ratio of other diffracted light is changed according to Embodiment 1. [Figure 12] FIG. 12 is a diagram schematically showing the configuration of an optical unit according to Embodiment 2. [Figure 13] FIG. 13 is a side view schematically showing the configuration of a specimen processing system according to Embodiment 3. [Figure 14] FIG. 14 is a diagram schematically showing the configuration of an optical system included in another specimen analyzer according to Embodiment 3. [Figure 15] FIG. 15 is a diagram schematically showing an optical system according to the prior art. MODE FOR CARRYING OUT THE INVENTION

[0012] <Embodiment 1> FIG. 1 is a block diagram showing the configuration of a specimen analyzer 1.

[0013] The sample analyzer 1 comprises a control unit 10, a calculation unit 11, a storage unit 12, a display unit 13, an input unit 14, a transport unit 15, a reading unit 16, a communication unit 17, and a measurement unit 20.

[0014] The control unit 10 is composed of a CPU. The arithmetic unit 11 is composed of a GPU. The storage unit 12 is composed of ROM, RAM, SSD, HDD, etc. The control unit 10 receives signals output by each part of the sample analyzer 1 and controls each part of the sample analyzer 1.

[0015] Furthermore, the control unit 10 executes a program stored in the memory unit 12 for cell analysis, and instructs the calculation unit 11 to perform cell analysis using an AI algorithm based on the detection signal obtained by the measurement unit 20. In this case, the AI ​​algorithm is a deep learning algorithm. More specifically, the calculation unit 11 executes an AI algorithm 32 (see Figure 2) composed of a trained neural network, and obtains the cell classification result based on the detection signal obtained by the measurement unit 20.

[0016] Furthermore, the arithmetic unit 11 may execute an AI algorithm 31 (see Figure 2) composed of a neural network before training, perform training using training data, and generate an AI algorithm 32 (see Figure 2) composed of a neural network before performing analysis using the AI ​​algorithm 32 composed of a trained neural network.

[0017] In Figure 1, the control unit 10 controls each part of the sample analyzer 1, and the calculation unit 11 performs cell classification using the AI ​​algorithm 32. However, a single control unit 10 may also control each part of the sample analyzer 1 and perform cell classification using the AI ​​algorithm 32.

[0018] The display unit 13 is comprised of a liquid crystal display. The input unit 14 is comprised of a pointing device including a keyboard, mouse, and touch panel. The liquid crystal display of the display unit 13 and the touch panel of the input unit 14 may be integrally configured. The reading unit 16 is comprised of a barcode reader. The reading unit 16 reads the barcode from the label attached to the sample container and obtains the sample ID. The transport unit 15 transports the sample rack holding the sample containers and transports it to the reading position of the reading unit 16. The communication unit 17 is a communication interface based on the Ethernet standard. The control unit 10 can communicate with other devices via the communication unit 17.

[0019] The measurement unit 20 includes a sample preparation unit 21, a fluid adjustment unit 22, an optical system 100, an amplifier 23, and an A / D converter 24.

[0020] The sample preparation unit 21 comprises a pipette for aspirating a sample from a sample container and a chamber for mixing the sample and reagents to prepare the sample. The sample used in the sample analyzer 1 can be any sample containing cells, such as blood, cerebrospinal fluid, bone marrow fluid, and body cavity fluids. The reagents mixed with the sample include a hemolytic agent for lysing red blood cells and a fluorescent dye for staining specific parts of cells. The fluid adjustment unit 22 comprises a container for storing sheath fluid and a syringe for aspirating the sample and sheath fluid. The fluid adjustment unit 22 supplies the sample and sheath fluid to the optical system 100 and adjusts the flow of the sample in the optical system 100. The optical system 100 measures the sample supplied by the fluid adjustment unit 22 and outputs an analog signal that reflects the characteristics of the cells. The amplifier 23 amplifies the analog signal output from the optical system 100. The A / D converter 24 converts the analog signal amplified by the amplifier 23 into a detection signal, which is a digital signal.

[0021] Figure 2 is a schematic diagram showing AI algorithm 31 before training and AI algorithm 32 after training.

[0022] As shown in the upper part of Figure 2, the training detection signal 41 used to train the AI ​​algorithm 31 before training is, for example, a detection signal obtained by measuring a specific cell with the sample analyzer 1. The type of cell (in the example in the upper part of Figure 2, "Cell A") obtained by another analyzer is associated with this detection signal.

[0023] The AI ​​algorithm 31 is composed of a neural network including multiple hidden layers. In this case, the neural network is, for example, a convolutional neural network having convolutional layers. The AI ​​algorithm 31 has an input layer 31a, an output layer 31b, and a hidden layer 31c. The AI ​​algorithm 31 is trained when a data set of detection signals obtained by sampling an analog signal obtained from one cell at a predetermined sampling period is input to the input layer 31a, and label values ​​corresponding to the cell type are input to the output layer 31b. By repeatedly performing this training in advance, the trained AI algorithm 32 is generated.

[0024] As shown in the lower part of Figure 2, the trained AI algorithm 32 also has an input layer 32a, an output layer 32b, and an intermediate layer 32c. The detection signal 42 obtained based on the subject's sample is input to the input layer 32a. As a result, classification information 43 regarding the type of cell corresponding to the detection signal 42 is output from the output layer 32b. The classification information 43 includes the probability that the target cell belongs to each of several types. Furthermore, based on the calculation results including these probabilities calculated by the calculation unit 11, the control unit 10 determines the type with the highest probability (in the example in the lower part of Figure 2, "Cell A") as the type of target cell and displays the determination result on the display unit 13.

[0025] Training of AI algorithm 31 and classification using AI algorithm 32 are performed by inputting a data set of detection signals obtained for each individual cell by one or more of the three light-receiving units 123, 133, and 143 (see Figure 3) as input data into the input layer 32a. Specifically, if a detection signal obtained from any one of the light-receiving units 123, 133, and 143 is used to obtain n data sets from the detection signals obtained for each individual cell, the number of detection signal data points input to AI algorithms 31 and 32 corresponding to one cell will be n, and the number of nodes in input layers 31a and 32a will also be n. For example, if three data sets of detection signals obtained from each of the three light-receiving units 123, 133, and 143 (see Figure 3) are input as input data into input layers 31a and 32a, 3n data sets will be obtained from the three detection signals, and the number of nodes in input layers 31a and 32a will also be 3n.

[0026] Figure 3 is a schematic diagram showing the configuration of the optical system 100. For convenience, the X, Y, and Z axes, which are orthogonal to each other, are indicated in Figure 3. The positive Z-axis direction is the flow direction of the sample in the flow cell 101.

[0027] The optical system 100 comprises a flow cell 101, a light source 111, an illumination optical system IS, focusing lenses 121, 131, 141, optical filters 122, 132, 142, and light receiving units 123, 133, 143. The illumination optical system IS comprises a collimator lens 112, cylindrical lenses 113A, 113B, a diffractive optical element (DOE) 114, and a focusing lens 115. The illumination optical system IS irradiates the flow cell 101 with illumination light.

[0028] The light source 111 is, for example, a semiconductor laser light source. The light source 111 emits light of a predetermined wavelength λ10 in the positive X-axis direction. The wavelength λ10 is, for example, 405 nm. The fast axis direction and slow axis direction of the light source 111 are parallel to the Y axis direction and the Z axis direction, respectively.

[0029] The collimator lens 112 has a curved surface that curves with respect to the Y and Z axes, reducing the divergence angle of the light emitted from the light source 111.

[0030] The cylindrical lens 113A has a lens surface that curves only in a direction parallel to the XY plane, and the generatrix of this lens surface is parallel to the Z axis. The cylindrical lens 113A focuses the light emitted from the light source 111 in the direction of the fast axis, adjusting the spread of the light in the fast axis direction to a nearly parallel state. In other words, the cylindrical lens 113A has the effect of making the light emitted from the light source 111 parallel only in the direction of the fast axis.

[0031] The cylindrical lens 113B has a lens surface that curves only in a direction parallel to the XZ plane, and the generatrix of this lens surface is parallel to the Y axis. The cylindrical lens 113B focuses the light emitted from the light source 111 in the direction of the slow axis, adjusting the spread of light in the slow axis direction to a nearly parallel state. In other words, the cylindrical lens 113B has the effect of making the light emitted from the light source 111 parallel only in the direction of the slow axis.

[0032] The collimator lens 112 and cylindrical lenses 113A and 113B are positioned such that the light emitted from the light source 111 and transmitted through them forms a nearly perfect circle when viewed in the X-axis direction. As a result, the light incident on the diffractive optical element 114 forms a nearly perfect circle.

[0033] The diffractive optical element 114 has a diffraction pattern formed on it with a complex uneven shape, such as grooves and inclinations, to impart a diffraction effect to the incident light. The diffractive optical element 114 can be manufactured, for example, based on the description in Japanese Patent Publication No. 5849954. The diffractive optical element 114 diffracts the light incident in the X-axis direction from the cylindrical lens 113B side along the X-axis direction, generating multiple diffracted lights with different propagation directions. These multiple diffracted lights are the spectrally separated portions of the incident light. The diffraction orders of the multiple diffracted lights are different from each other. The focusing lens 115 focuses the multiple diffracted lights generated from the diffractive optical element 114 onto the flow cell 101. The multiple diffracted lights generated in the diffractive optical element 114 with different propagation directions are focused onto the flow cell 101 to form illumination light. Of the multiple diffracted lights included in the illumination light, zero-order diffracted light is generated in the direction normal to the diffractive optical element 114, and other diffracted lights other than the zero-order diffracted light are generated in a direction different from the normal direction of the diffractive optical element 114. The intensities of the diffracted light other than the zero-order diffracted light are approximately the same. On the other hand, the intensity of the zero-order diffracted light is not equal to that of the other diffracted light, although this depends on the design of the diffracting optical element 114. Here, light intensity refers to the amount of light (mW) of each diffracted light at the focusing surface of the illumination light.

[0034] A sample containing cells such as neutrophils, eosinophils, basophils, lymphocytes, and monocytes flows through flow cell 101. Illumination light is shone on the cells flowing through flow cell 101, and each diffracted light in the illumination light generates forward scattered light, side scattered light, and fluorescence from the irradiated part of the cell. Forward scattered light is generated in the positive X-axis direction, while side scattered light and fluorescence are generated in a direction intersecting the X-axis (for example, the Y-axis direction). Here, when light of wavelength λ10 is shone on a fluorescent dye used to stain cells, fluorescence of wavelength λ11 is generated from this fluorescent dye.

[0035] The focusing lens 121 focuses the forward scattered light generated from the cells onto the light receiving unit 123. The optical filter 122 is configured to transmit only light with a wavelength of λ10. The light receiving unit 123 receives the forward scattered light that has passed through the optical filter 122 and outputs a detection signal according to the light reception intensity. The light receiving unit 123 is a photomultiplier tube (PMT).

[0036] The focusing lens 131 focuses the side-scattered light generated from the cells onto the light-receiving unit 133. The optical filter 132 is configured to transmit only light with a wavelength of λ10. The light-receiving unit 133 receives the side-scattered light that has passed through the optical filter 132 and outputs a detection signal according to the light detection intensity. The light-receiving unit 133 is a photomultiplier tube (PMT).

[0037] The focusing lens 141 focuses the fluorescence emitted from the cells onto the light-receiving unit 143. The optical filter 142 is configured to transmit only light of wavelength λ11. The light-receiving unit 143 receives the fluorescence transmitted through the optical filter 142 and outputs a detection signal corresponding to the light-receiving intensity. The light-receiving unit 143 is a photomultiplier tube (PMT). Photodiodes may be used instead of photomultiplier tubes (PMTs) in the light-receiving units 123, 133, and 143.

[0038] Figure 4 schematically shows the flow cell 101 and the illumination light. Figure 4 includes the same X, Y, and Z axes as in Figure 3.

[0039] Inside the flow cell 101, a channel 101a through which the sample flows is formed parallel to the Z-axis. By flowing the sheath fluid along with the sample through channel 101a, the cells contained in the sample are enveloped in the sheath fluid and pass through the central region CE of channel 101a. Illumination light focused by the focusing lens 115 is irradiated onto a predetermined irradiation area R located in the central region CE of channel 101a. The sample concentration and sample flow rate are adjusted so that only one cell is positioned in the irradiation area R at a time, in other words, so that two or more cells do not pass through the irradiation area R simultaneously.

[0040] The lower part of Figure 4 shows an image obtained by illuminating a darkroom with illumination light generated by a prototype diffractive optical element 114 (AGC Inc.) and capturing the image with a camera (Basler AG, acA3800).

[0041] In the illumination light image in Figure 4, the black areas indicate areas without light, and the white dots indicate areas with light. The white dot near the center of the illumination light image indicates the 0th order diffracted light generated by the diffracting optical element 114. The other white dots in the illumination light image indicate diffracted light other than the 0th order diffracted light generated by the diffracting optical element 114. The diffracted light in this embodiment includes 0th order diffracted light, +1 to +300th order diffracted light, and -1 to -300th order diffracted light, and is shown in the illumination light image as a total of 601 white dots. Diffracted patterns (steps or grooves) are formed on the diffracting optical element 114 so that each type of diffracted light is distributed as shown in the illumination light image in Figure 4.

[0042] Figure 5 schematically shows the distribution pattern of diffracted light contained in illumination light.

[0043] Figure 5 shows an image of the illumination area R divided into a grid by multiple squares with sides the same length as the diameter of the zero-order diffracted light spot. The hatched squares in the center indicate the region containing the zero-order diffracted light spot, the black squares indicate the region containing the spot of one other diffracted light besides the zero-order diffracted light, and the white squares indicate the region not containing the diffracted light spot. In Figure 5, cells passing through the illumination area R are shown as dashed circles. Since the diameter of the diffracted light spot contained in the illumination light image in Figure 4 is approximately 1 μm, in this case, the size of each square is 1 × 1 μm. The size of the cells is approximately 10 μm.

[0044] The size of the illumination light and its length in the Y-axis or Z-axis direction within the irradiation area R (see Figure 4) can be expressed in terms of pixels, assuming each region of the grid is considered as 1 pixel. In the example shown in Figure 5, the length of the illumination light in the flow direction of the sample in the flow cell 101 (Z-axis direction) is px1 (pixel), the length of the illumination light in the short direction (Y-axis direction) is px2 (pixel), and the size of the illumination light is px1 × px2 (pixels).

[0045] The diffractive optical element 114 is designed so that the multiple diffracted light beams constituting the illumination light are distributed in a predetermined pattern. In this case, the predetermined pattern is a random pattern. The pattern may not have any repetition of a particular pattern, or it may have periodicity in which a particular pattern is repeated. However, it is preferable that at least one other diffracted light beam is arranged in a region that is 1 pixel long in the Y-axis direction and extends in the Z-axis direction, so that the entire cell region is exposed to the illumination light at least once.

[0046] When a sample is flowed into the flow channel 101a of the flow cell 101 during measurement, the cells in the sample move in the positive Z-axis direction within the irradiation range R of the illumination light. At this time, the fluid adjustment unit 22 adjusts the flow velocity to be approximately constant. When diffracted light contained in the illumination light is irradiated onto cells moving in the positive Z-axis direction, forward scattered light and side scattered light are generated from the part of the cell irradiated with diffracted light. Also, when diffracted light is irradiated onto cells stained with a fluorescent dye, fluorescence is generated from the fluorescent dye irradiated with diffracted light. The light receiving unit 123 receives forward scattered light generated by one or more diffracted lights irradiated onto the cells, the light receiving unit 133 receives side scattered light generated by one or more diffracted lights irradiated onto the cell location, and the light receiving unit 143 receives fluorescence generated by one or more diffracted lights irradiated onto a predetermined location of the stained cell.

[0047] As cells flow in the positive Z-axis direction, the number of diffracted light beams irradiated onto the cells changes, and the location of each diffracted light beam that hits the cell changes. As a result, the intensity of forward scattered light, side scattered light, and fluorescence emitted from the cells changes over time. Consequently, the detection signals from each light-receiving unit 123, 133, and 143 also change over time. The calculation unit 11 classifies the cells using the AI ​​algorithm 32 based on these detection signals.

[0048] In conventional devices, such as those exemplified in Patent Document 1, as explained with reference to Figure 15, diffraction patterns including 0th and higher-order diffracted light generated from diffracting optical elements were blocked by lenses 1002 and 1003 and a spatial filter. However, this configuration was a constraint on miniaturizing the optical system. In contrast, after repeated studies by the inventors, they found that by irradiating cells with illumination light containing 0th-order diffracted light, whose relative intensity to other diffracted light is 10 times or less, it is possible to enable cell classification by AI algorithms while simultaneously achieving miniaturization of the device.

[0049] Here, we define the RI as the relative intensity of the zero-order diffracted light with respect to at least one other diffracted light, and we will explain how to calculate the relative intensity RI.

[0050] Hereinafter, the intensity of the incident light incident on the diffractive optical element 114 will be defined as L0. The intensity of the 0th-order diffracted light will be defined as L1. The intensity of the other diffracted light other than the 0th-order diffracted light will be defined as L2. The light intensities L0, L1, and L2 can be measured using an optical power meter, or obtained by imaging the illumination light using a beam profiler or camera and integrating the brightness values ​​of multiple pixels corresponding to the imaged light. If the intensities of the other diffracted light are substantially the same, for example, with a variation of 3% or less, the intensity L2 can be the intensity of any one of the other diffracted light. Alternatively, if they are not substantially the same, representative values ​​such as the mean, median, mode, or maximum of several of the other diffracted light may be used.

[0051] Furthermore, let R1 be the ratio of the intensity L1 of the zero-order diffracted light generated from the diffracting optical element 114 to the intensity L0 of the incident light. Let R2 be the diffraction efficiency of the diffracting optical element 114. Let N be the number of other diffracted light rays included in the illumination light. The ratios R1 and R2 are characteristic values ​​of the diffracting optical element. N is the number of points (also called spots) of illumination light formed by other diffracted light rays other than the zero-order diffracted light. The intensity L1 of the zero-order diffracted light is expressed by the following equation (11), and the intensity L2 of the other diffracted light rays is expressed by the following equation (12).

[0052] L1 = L0 × R1 …(11) L2 = (L0 - L1) × R2 / N …(12)

[0053] The relative intensity RI is expressed by the following equation (13).

[0054] RI=L1 / L2=N×R1 / (R2-R1×R2) …(13)

[0055] From this equation (13), the relative intensity RI can be determined from the measured values ​​obtained by actually measuring the light intensities L1 and L2, or it can be determined from the characteristic values ​​R1 and R2 of the diffractive optical element 114 and the number of spots N.

[0056] Let S (in units of pixels) be the total area of ​​the region where spots can be formed, assuming that one spot in the illumination light is one pixel. Let RS (in units of % (number of spots / number of pixels)) be the ratio of the total area to the area where illumination light points (spots) formed by other diffracted light are formed. In other words, the spot ratio RS is the ratio of the number of regions containing other diffracted light to the number of regions in a grid (see Figure 5) that contain regions containing one diffracted light and regions that do not contain diffracted light. In this case, the number of spots N is expressed by the following equation (14).

[0057] N = S × RS / 100 …(14)

[0058] According to equations (11) to (14) above, the following can be said.

[0059] The relative intensity RI can be reduced by decreasing the ratio R1 of the zero-order diffracted light to the incident light. The ratio R1 is preferably less than 3%, for example.

[0060] By increasing the diffraction efficiency R2 of the diffractive optical element 114, the relative intensity RI can be reduced. The diffraction efficiency R2 is preferably, for example, 60% or higher.

[0061] The relative intensity RI can be reduced by decreasing the number of spots N contained in the illumination light. In other words, the relative intensity RI can be reduced by decreasing the number of spots N by decreasing the area S of the illumination light or by decreasing the spot ratio RS. The lengths in the Y-axis and Z-axis directions that form the area S are set to, for example, 59 pixels and 999 pixels, respectively. The preferred range for the length in the Z-axis direction that constitutes the area S will be explained later with reference to Figure 8. The spot ratio RS is preferably set to, for example, 0.3% or more and 2% or less. The preferred range for the spot ratio RS will be explained later with reference to Figure 11.

[0062] However, the preferred range is not limited to the above, as long as the relative intensity RI can be reduced to 10 times or less.

[0063] The following explanation, with reference to Figures 6-10, describes how the effect of zero-order diffracted light can be effectively suppressed when the relative intensity RI is 10 times or less.

[0064] First, the inventor investigated the acceptable relative intensity RI for the optical system while maintaining the cell classification performance of the AI ​​algorithm, using a prototype device (hereinafter referred to as "the example"). The diffractive optical element 114 used in the example was the prototype whose illumination light image was shown in the lower part of Figure 4. The specifications of the illumination optical system IS, including this prototype diffractive optical element 114, were as follows.

[0065] [Table 1]

[0066] The prototype diffractive optical element 114 had a transmissive configuration that transmitted incident light and emitted it. The ratio of the intensity of the 0th-order diffracted light to the incident light, R1, was 0.1%. The diffraction efficiency R2 of the diffractive optical element 114 was 0.7. The illumination light size was 999 pixels in the direction of sample flow and 59 pixels in the direction perpendicular to the sample flow. The spot ratio RS of other diffracted light was 1.09%. The relative intensity RI of the 0th-order diffracted light to other diffracted light was 0.93 times.

[0067] Figures 6 and 7 are graphs showing the detection signals obtained in experiments conducted to determine the appropriate illumination intensity.

[0068] When classifying leukocytes, detection signals are obtained from light emitted by cells such as neutrophils, eosinophils, basophils, lymphocytes, and monocytes. However, the intensity of the detection signal based on lymphocytes is the lowest, and the intensity of the detection signal based on neutrophils is the highest. Therefore, in order to acquire the detection signal of lymphocytes in distinction from noise, it is necessary to set the intensity of the illumination light, including zero-order diffracted light, to a certain extent, while not increasing the intensity of the illumination light beyond a certain point so that the light receiving unit does not become saturated by the light based on neutrophils. Below, the inventors conducted an experiment to measure leukocytes using the apparatus of the example. In this experiment, control blood containing lymphocytes and neutrophils (streck, CD-Chex Plus, catalog no. 213367) was used as a sample, and detection signals based on lateral scattered light generated from each cell upon irradiation with illumination light were acquired by the light receiving unit 133 for multiple types of cells contained in this sample. In addition, the sample was fluorescently labeled to identify the cell types of lymphocytes and neutrophils, and detection signals based on fluorescence were acquired by the light receiving unit 143.

[0069] In the graph shown in Figure 6, the value of the detection signal when the light-receiving unit 133 is saturated is set to 1.0, the relative value of the detection signal is shown on the vertical axis, and the measurement time [μsec] is shown on the horizontal axis. The upper graph in Figure 6 shows the detection signal obtained when lymphocytes were actually measured. The lower graph in Figure 6 is an enlarged view of the upper graph in Figure 6. In the lower graph in Figure 6, the range of the two dashed lines is the range in which noise superimposed on the detection signal is approximately included. The upper limit of the noise range, i.e., the noise threshold, is set, for example, to the value obtained by adding the 2SD value of the noise (0.00856) to the baseline (0) in the noise region near the left edge of the graph. In the lower graph in Figure 6, the area enclosed by the dashed line is the smallest detection signal among the detection signals excluding noise. Therefore, the area enclosed by the dashed line can be considered as a detection signal based on one of the other diffracted light rays other than the 0th-order diffracted light contained in the illumination light. The value of this detection signal is approximately 0.02.

[0070] If the sensitivity of the light-receiving unit 133 or the intensity of the illumination light is reduced, the lymphocyte detection signal will fall below the noise threshold, making it impossible to distinguish between the lymphocyte detection signal and the noise detection signal. Therefore, in Figure 6, the intensity of the illumination light of the light-receiving unit 133 is set to a level at which the lymphocyte detection signal can be acquired in distinction from noise. Specifically, the sensitivity of the light-receiving unit 133 and the intensity of the illumination light are set so that the detection signal based on one of the other diffracted light rays enclosed by the dashed line slightly exceeds the noise threshold.

[0071] The upper graph in Figure 7 shows the detection signal obtained when neutrophils were actually measured. In this case, the detection signal was acquired with the same sensitivity of the light-receiving unit 133 and illumination light intensity as when lymphocytes were detected in Figure 6. In the upper graph of Figure 7, the area enclosed by the dashed line is the smallest detection signal among the detection signals excluding noise. Therefore, the area enclosed by the dashed line can be considered as the detection signal for neutrophils based on one of the diffracted rays contained in the illumination light. The value of this detection signal is approximately 0.1.

[0072] As mentioned above, the relative intensity RI of the zero-order diffracted light in the diffracting optical element 114 used in this experiment is 0.93 times, so the value of the detection signal based on the zero-order diffracted light contained in the illumination light is about the same as the value of the detection signal based on one of the other diffracted lights. Also, in this case, the value of the detection signal when the light receiving unit 133 becomes saturated is 1.0. Therefore, if the value of the detection signal based on the zero-order diffracted light is in the range of 0.1 to 1.0, it is possible to prevent saturation of the light receiving unit 133. That is, if we assume zero-order diffracted light with a relative intensity RI 10 times that of the upper graph in Figure 7, the detection signal based on the zero-order diffracted light extends to around 1.0, as shown by the dotted line in the lower graph in Figure 7. However, since the detection signal based on the zero-order diffracted light at this time is less than or equal to the value when the light receiving unit 133 becomes saturated (1.0), saturation of the light receiving unit 133 is avoided.

[0073] As described above, according to the experiments shown in Figures 6 and 7, by setting the relative intensity RI to 10 times or less in the classification of white blood cells, the detected signal can be distinguished from noise in both the case of lymphocytes, which have the smallest detection signal, and the case of neutrophils, which have the largest detection signal, and the saturation of the light receiving unit 133 can be avoided.

[0074] In this experiment, we examined the detection signal of the photodetector 133 based on lateral scattered light, but the same applies to detection signals based on forward scattered light and fluorescence. That is, in both the detection signal of the photodetector 123 that receives forward scattered light and the detection signal of the photodetector 143 that receives fluorescence, by setting the relative intensity RI to 10 times or less, the detection signal can be distinguished from noise in both lymphocytes and neutrophils, and saturation of the photodetector 133 can be avoided.

[0075] Next, referring to Figure 8, we will explain the experimental results regarding the classification performance of the AI ​​algorithm when the length of the illumination light along the Z-axis is changed.

[0076] In this experiment, the apparatus described in the example was used. The spot ratio RS of other diffracted light of the prototype diffracting optical element 114 was 1.09%. The relative intensity RI of the 0th-order diffracted light of the diffracting optical element 114 was 0.93 times. A sample containing neutrophils, eosinophils, basophils, lymphocytes, and monocytes was flowed through the flow cell 101, and illumination light with a Z-axis length of 999 pixels was irradiated onto the flow cell 101. Each cell was classified using the AI ​​algorithm 32 with the detection signal of the light receiving unit 133 based on lateral scattered light. The correctness of the classification result for each cell was determined and the F-value was obtained, and the average F-value of each cell was obtained as the classification performance. In addition, from the entire set of detection signals corresponding to illumination light with a Z-axis length of 999 pixels, detection signals corresponding to illumination light with Z-axis lengths of 100, 200, 300, 400, 500, 600, and 700 pixels were extracted, and the above F-value and the average F-value were obtained for each extracted detection signal.

[0077] Figure 8 is a graph showing the classification performance when the length of the illumination light is changed. In the graph in Figure 8, the vertical axis represents the classification performance, and the horizontal axis represents the length of the illumination light in the Z-axis direction (the length of the illumination light corresponding to the extracted detection signal). The table in the lower part of Figure 8 shows the details of the graph in the upper part of Figure 8.

[0078] As shown in the graph in Figure 8, with the optical system used in this embodiment, by setting the relative intensity RI to 1x or less, an accuracy rate of over 95% was achieved for cell classification performance by the AI ​​algorithm in all cases where the Z-axis length was 100 pixels or more. The classification performance improved as the Z-axis length of the illumination light increased. This is thought to be because the amount of information in the detection signal that can be acquired increases as the Z-axis length of the illumination light increases, allowing for more accurate cell classification. Furthermore, the classification performance when the Z-axis length of the illumination light exceeds 300 pixels is one step higher than the classification performance when the length is 100 or 200 pixels, and the change in classification performance is smaller. Therefore, it is more preferable to set the Z-axis length of the illumination light to 300 pixels or more.

[0079] In this experiment, we investigated the length of the illumination light when using the detection signal of the light-receiving unit 133 based on lateral scattered light, but the same applies when using the detection signal based on forward scattered light and fluorescence. That is, in both cases, when using the detection signal of the light-receiving unit 123 that receives forward scattered light and when using the detection signal of the light-receiving unit 143 that receives fluorescence, it is assumed that it is preferable to set the length of the illumination light in the Z-axis direction to 300 pixels or more.

[0080] Next, the acceptable range of relative intensity radioisotopes (RI) for maintaining cell classification performance was confirmed through simulation. Figures 9 and 10 are graphs showing the simulation results examining the differences in the waveform of the detection signal when the relative intensity RI is different. In the simulation, the relative intensity RI of the zero-order diffracted light was changed to 46 times, 10 times, 5 times, and 1 time.

[0081] In this simulation, first, a map was created that reproduced the same diffracted light distribution pattern as the image shown in the lower part of Figure 4 using pixel values. Specifically, a grid-like map as shown in Figure 5 was created using spreadsheet software, and 1 was entered for the grid corresponding to the white light spots in the image of Figure 4, and 0 for the black areas. For the grid corresponding to the 0th order diffracted light, 46, 10, 5, or 1 was entered depending on the relative intensity RI. Next, a pre-created bead image was superimposed on this map, and the dot product of the pixel values ​​of the grid in the overlapping area and the pixel values ​​of the bead image was calculated and used as the value of the detection signal. This calculation was repeated each time the position of the bead image was moved from left to right by one pixel at a time step, creating a time-series waveform of the detection signal. The graphs in Figures 9 and 10 show the waveform of the detection signal changing over time. In each graph, the vertical axis shows the intensity of the detection signal, and the horizontal axis shows the number of elapsed steps corresponding to the elapsed time. The left and right ends of the graphs show the detection signals acquired when the beads are positioned at the left and right ends of the irradiation range R, respectively. In other words, the elapsed time on the horizontal axis corresponds to the position of the cell moving in the positive Z-axis direction.

[0082] The upper graph in Figure 9 shows the detection signal when the relative intensity RI of the 0th-order diffracted light is set to 46 times, that is, the detection signal when the intensity value of the 0th-order diffracted light is 46 and the intensity values ​​of the other diffracted light are 1. In this case, the intensity of the light produced by the 0th-order diffracted light is significantly higher than the intensity of the light produced by the other diffracted light, so a significantly high-intensity detection signal is detected at the moment the 0th-order diffracted light irradiates the beads. If the detector and subsequent amplifiers and A / D conversion circuits are configured to detect such a significantly high-intensity detection signal, the signal resolution of the low-intensity detection signals from the other diffracted light will be greatly reduced when the detection signal is sampled from the analog signal and converted to a digital signal. Specifically, if the total gradation of the digital signal is 256 gradations, the high-value 5-256 gradations will only be used for the detection signal from the 0th-order diffracted light, and the detection signals from the other diffracted light will only use an extremely narrow range of gradations, such as the low-value 1-5 gradations. Therefore, it becomes difficult to classify cells using AI algorithms that use the 0th-order diffracted light and other diffracted light.

[0083] The lower graph in Figure 9 shows the detection signal when the relative intensity RI is 10 times greater, that is, when the intensity value of the 0th-order diffracted light is 10 and the intensity values ​​of the other diffracted light are 1. In this case, compared to the upper graph in Figure 9, the difference between the intensity of the light produced by the 0th-order diffracted light and the intensity of the light produced by the other diffracted light has narrowed. By configuring the detector and subsequent amplifiers and A / D conversion circuits to detect a detection signal with a relative intensity of about 10 times, the signal resolution of the low-intensity detection signal from the other diffracted light is maintained to a certain extent even when the detection signal is sampled from the analog signal and converted to a digital signal. Specifically, if the total gradation of the digital signal is 256 gradations, the high-value 26-256 gradations are used only for the detection signal from the 0th-order diffracted light, but the detection signal from the other diffracted light can use gradations such as the low-value 1-25 gradations. If there is a signal resolution of about 25 gradations for the other diffracted light of the 0th-order diffracted light, it becomes possible to classify cells with a certain level of accuracy using an AI algorithm.

[0084] The upper graph in Figure 10 shows the detection signal when the relative intensity RI is 5 times higher, that is, when the intensity value of the 0th-order diffracted light is 5 and the intensity value of the other diffracted light is 1. In this case, compared to the lower graph in Figure 9, the difference between the intensity of the light produced by the 0th-order diffracted light and the intensity of the light produced by the other diffracted light has narrowed further. If the detector and subsequent amplifiers and A / D conversion circuits are configured to detect a detection signal with a relative intensity of about 5 times, then if the total gradation of the digital signal is 256 gradations, the high-value gradations from 51 to 256 are used only for the detection signal from the 0th-order diffracted light, while the detection signal from the other diffracted light can use gradations from the low-value gradations, such as 1 to 50. Therefore, it becomes possible to classify cells with higher accuracy using an AI algorithm.

[0085] The lower graph in Figure 10 shows the detection signal when the relative intensity RI is set to 1, that is, the detection signal when the intensity values ​​of the 0th-order diffracted light and other diffracted light are set to 1. By configuring the detector and subsequent amplifiers and A / D conversion circuits to detect a detection signal with a relative intensity of approximately 1, if the total grayscale of the digital signal is 256, all 256 grayscale levels can be used for the detection signal from other diffracted light. Therefore, it becomes possible to classify cells with even higher accuracy using an AI algorithm.

[0086] Next, referring to Figure 11, we will explain the simulation results of the performance of the diffractive optical element 114 when the spot ratio RS of other diffracted light is changed.

[0087] In this simulation, the measurement target was the fluorescence image of a bead, and the relative intensity RI of the zero-order diffracted light was set to 1. The spot ratios RS of the other diffracted lights were set to 0.1%, 0.3%, 0.5%, 0.7%, 1%, 2%, and 3%. For each spot ratio RS, 1000 types of diffracting optical elements 114 that produce illumination light with different distribution patterns were used to generate a detection signal for the light-receiving unit 143 based on fluorescence from the measurement target (bead) and the distribution pattern of the illumination light. The original bead image (fluorescence image) was reconstructed from the generated detection signal and the distribution pattern of the illumination light, and the PSNR (peak signal-to-noise ratio) was calculated as an index corresponding to the signal amount of the detection signal. PSNR is an index that shows how accurately the bead image can be reconstructed, and it is expected that cell classification will be more accurate in ghost cytometry by using a detection signal with a high PSNR.

[0088] Figure 11 is a graph showing the maximum PSNR for each spot ratio RS. In the graph in Figure 11, the horizontal axis represents the spot ratio RS, and the vertical axis represents the maximum PSNR based on 1000 types of diffractive optical elements 114.

[0089] As shown in the graph in Figure 11, the maximum PSNR when the spot ratio RS is between 0.3% and 2% was significantly larger than the maximum PSNR when the PSNR was between 0.1% or 3%. This indicates that when using the diffractive optical element 114 that yields the maximum PSNR, setting the spot ratio RS to between 0.3% and 2% effectively increases the signal strength of the detection signal, allowing for accurate reconstruction of the bead image. Furthermore, setting the spot ratio RS to 1% resulted in the highest possible maximum PSNR. Therefore, it is preferable for the spot ratio RS to be set between 0.3% and 2%, and even more preferable for it to be 1%.

[0090] <Effects of the optical system and sample analyzer according to Embodiment 1> As shown in Figure 3, the illumination optical system IS includes a diffractive optical element 114 to which light from the light source 111 is incident, and illuminates the flow cell 101 with illumination light (see Figures 4 and 5) in which multiple diffracted light, including zero-order diffracted light, generated by the diffractive optical element 114 is distributed in a predetermined distribution pattern. The light receiving units 123, 133, and 143 receive light generated from the cells flowing through the flow cell 101 by illumination light containing multiple diffracted light, including zero-order diffracted light. The illumination optical system IS does not have a configuration to block zero-order and higher-order diffracted light.

[0091] This section explains the miniaturization effect achieved by not including a configuration to block 0th and higher-order diffracted light. In the cell analysis device described in Patent Document 1, two lenses are provided between the diffractive optical element and the objective lens to temporarily image the structured illumination in order to block diffraction patterns that include 0th and higher-order diffracted light. The focal lengths of the two lenses used in Patent Document 1 are each 150 mm. In other words, in Patent Document 1, a distance of 300 mm must be secured just for the distance between the two lenses between the diffractive optical element and the objective lens. On the other hand, in the experimental device prototyped by the inventors, the distance between the diffractive optical element 114 and the focusing lens 115 shown in Figure 3 is approximately 20 mm, and the distance between the DOE and the objective lens was reduced to less than 1 / 15 compared to the optical system in Patent Document 1. Furthermore, while the distance between lenses in Patent Document 1 is 300 mm, the size of the entire optical system of the experimental device in the X-axis direction is approximately 350 mm, which also indicates that significant miniaturization has been achieved.

[0092] <Embodiment 2> The optical system 100 of Embodiment 2 further includes a configuration for receiving light generated when other light that does not pass through the diffractive optical element 114 is irradiated onto the cells.

[0093] Figure 12 is a schematic diagram showing the configuration of the optical system 100 according to Embodiment 2.

[0094] The optical system 100 of Embodiment 2, compared to the optical system 100 of Embodiment 1 shown in Figure 3, includes a light source 151, a collimator lens 152, a light source 153, a collimator lens 154, a dichroic mirror 155, a cylindrical lens 156, a dichroic mirror 157, dichroic mirrors 124, 134, 144, optical filters 125, 135, 145, light receiving units 126, 136, 146, and a beam stopper 116. The dichroic mirror 157 is included in the illumination optical system IS. The differences from Embodiment 1 will be described below.

[0095] Light sources 151 and 153 are, for example, semiconductor laser light sources. Light source 151 emits light of a predetermined wavelength λ20 in the negative Y-axis direction, and light source 153 emits light of a predetermined wavelength λ30 in the positive X-axis direction. Wavelength λ20 is, for example, 642 nm, and wavelength λ30 is, for example, 488 nm. Collimator lenses 152 and 154 convert the light emitted from light sources 151 and 153 into parallel light, respectively. The dichroic mirror 155 transmits the light of wavelength λ20 from light source 151 and reflects the light of wavelength λ30 from light source 153. The dichroic mirror 155 aligns the optical axes of the light from light sources 151 and 153.

[0096] The cylindrical lens 156 focuses the light from the dichroic mirror 155 in the Z-axis direction, making it flattened at the position of the flow cell 101. The dichroic mirror 157 transmits light of wavelength λ10 from the light source 111 and reflects light of wavelength λ20 from the light source 151 and light of wavelength λ30 from the light source 153. The dichroic mirror 157 aligns the optical axes of the light from the light sources 151 and 153 with the central axis of the illumination light from the diffractive optical element 114.

[0097] The focusing lens 115 focuses the light from the light sources 111, 151, and 153 onto the flow channel 101a of the flow cell 101. The focusing lens 115 is configured to suppress chromatic aberration for light with wavelengths λ10, λ20, and λ30. The light from the light sources 151 and 153 is irradiated onto the flow channel 101a of the flow cell 101 in a flattened shape with a small width in the Z-axis direction, due to the action of the cylindrical lens 156.

[0098] When light from light sources 151 and 153 is shone onto cells flowing through flow cell 101, forward scattered light, side scattered light, and fluorescence are generated from the irradiated areas of the cells. Here, it is assumed that when light of wavelength λ30 is shone onto other fluorescent dyes used to stain cells, light of wavelength λ31 is generated from those other fluorescent dyes.

[0099] The focusing lens 121 focuses the forward scattered light of wavelength λ20 generated from the cells onto the light receiving unit 126. The dichroic mirror 124 reflects the forward scattered light of wavelength λ10 and transmits the forward scattered light of wavelength λ20. The beam stopper 116 blocks the light of wavelength λ20 that has passed through the flow cell 101 without irradiating the cells, and allows the forward scattered light of wavelength λ20 generated from the cells to pass through. The optical filter 125 is configured to transmit only light of wavelength λ20. The light receiving unit 126 receives the forward scattered light of wavelength λ20 that has passed through the optical filter 125 and outputs a detection signal according to the received light intensity. The light receiving unit 126 is, for example, a photodiode (PD).

[0100] The focusing lens 131 focuses the side-scattered light of wavelength λ10 generated from the cells onto the light-receiving unit 133 and focuses the side-scattered light of wavelength λ20 generated from the cells onto the light-receiving unit 136. The dichroic mirror 134 reflects the side-scattered light of wavelength λ10 and transmits the side-scattered light of wavelength λ20. The optical filter 135 is configured to transmit only light of wavelength λ20. The light-receiving unit 136 receives the side-scattered light of wavelength λ20 that has passed through the optical filter 135 and outputs a detection signal according to the received light intensity. The light-receiving unit 136 is, for example, a photodiode (PD).

[0101] The focusing lens 141 focuses fluorescence of wavelength λ11 emitted from the cells onto the light-receiving unit 143 and focuses fluorescence of wavelength λ31 emitted from the cells onto the light-receiving unit 146. The dichroic mirror 144 reflects fluorescence of wavelength λ11 and transmits fluorescence of wavelength λ31. The optical filter 145 is configured to transmit only light of wavelength λ31. The light-receiving unit 146 receives the fluorescence of wavelength λ31 that has passed through the optical filter 145 and outputs a detection signal according to the received light intensity. The light-receiving unit 146 is, for example, a photomultiplier tube (PMT).

[0102] In Embodiment 2, a scattergram and histogram are generated for each sample based on the detection signals from the light-receiving units 126, 136, and 146, and the cells are classified based on the generated scattergram and histogram.

[0103] <Effects of the optical system and sample analyzer according to Embodiment 2> As shown in Figure 12, the illumination optical system IS further includes a dichroic mirror 157 (matching optical element) that matches other light emitted from light sources 151 and 153 (other light sources) to the illumination light. The optical system 100 further includes dichroic mirrors 124, 134, and 144 (separation optical elements) that separate the light generated from cells by the illumination light from the light generated from cells by other light, and light receiving units 126, 136, and 146 (other light receiving units) that receive light based on the other light separated by the dichroic mirrors 124, 134, and 144 (separation optical elements).

[0104] With this configuration, the sample analyzer 1 can perform a variety of analyses because it can acquire further optical information of cells from other light emitted from light sources 151 and 153.

[0105] As shown in Figure 12, the light sources 151 and 153, and the configuration for receiving light generated from cells by the light from the light sources 151 and 153, are arranged in the gap between the light source 111 and the configuration for receiving light generated from cells by the light from the light source 111. Therefore, according to the optical system 100 of Embodiment 2, the optical system 100 can be made compact while having the above two types of configurations.

[0106] By simultaneously irradiating the flow channel 101a of the flow cell 101 with light from light source 111 and light from light sources 151 and 153, detection signals from light receiving units 123, 133, and 143, and detection signals from light receiving units 126, 136, and 146 can be acquired using a single sample. Furthermore, because these detection signals can be acquired simultaneously, analysis using the AI ​​algorithm 32 based on the detection signals from light receiving units 123, 133, and 143, and analysis using scattergrams and histograms based on the detection signals from light receiving units 126, 136, and 146 can be performed simultaneously. Therefore, the time required for the two types of analysis can be reduced.

[0107] <Embodiment 3> In Embodiment 2, a configuration for receiving light generated from cells using light from light source 111, and a configuration for receiving light generated from cells using light from light sources 151 and 153, were arranged in the optical system 100 of one sample analyzer 1. In contrast, in Embodiment 3, these two configurations are arranged in separate sample analyzers, and these two sample analyzers and other analyzers are connected by a transport device.

[0108] Figure 13 is a schematic side view showing the configuration of the sample processing system 5.

[0109] The specimen processing system 5 comprises specimen analyzers 1 and 2, a smear preparation device 3, and a transport device 4.

[0110] The specimen analyzer 1 of Embodiment 3 has the same configuration as the specimen analyzer 1 of Embodiment 1. The specimen analyzer 2 is the same as the specimen analyzer 2 of Embodiment 2, but the configuration for receiving light generated from cells by light from the light source 111 is omitted. Compared to the specimen analyzer 1 of Embodiment 2, the specimen analyzer 2 is equipped with an optical system 300 instead of an optical system 100. The optical system 300 will be described later with reference to Figure 14. The smear preparation device 3 is a device for preparing smears from blood specimens. The transport device 4 is arranged across the specimen analyzers 1 and 2 and the smear preparation device 3, and transports the specimen containers 202 held in the specimen rack 201 to each device.

[0111] The transport device 4 is operated by an operator to transport the sample rack 201, which is placed at the right end of the transport device 4, to the left, positioning the sample container 202 in front of the sample analyzers 1 and 2 and the smear preparation device 3.

[0112] Sample analyzer 1 reads the barcode from the label attached to the sample container 202 using the reader unit 16 (see Figure 1). Sample analyzer 1 measures the sample in the sample container 202 and classifies the cells using the AI ​​algorithm 32 based on the detection signal. Sample analyzer 2 reads the barcode from the label attached to the sample container 202 using the reader unit 16. Sample analyzer 2 measures the sample in the sample container 202 and classifies the cells based on the detection signal using a scattergram or histogram. Smear preparation device 3 reads the barcode from the label attached to the sample container 202 and prepares a smear from the sample in the sample container 202. The sample rack 201, transported to the left end of the transport device 4, is removed by the operator.

[0113] Figure 14 is a schematic diagram showing the configuration of the optical system 300 provided by the sample analyzer 2.

[0114] Compared to the optical system 100 of Embodiment 2 shown in Figure 12, the optical system 300 omits the light source 111, collimator lens 112, cylindrical lenses 113A and 113B, diffractive optical element 114, dichroic mirror 157, optical filters 122, 132, and 142, light receiving units 123, 133, and 143, and dichroic mirrors 124, 134, and 144. In addition, the light source 151 emits light in the positive X-axis direction, the light source 153 emits light in the positive Y-axis direction, and the direction of light propagation from the light sources 151 and 153 is aligned with the positive X-axis direction by the dichroic mirror 155.

[0115] <Effects of the optical system and sample analyzer according to Embodiment 3> According to Embodiment 3, the sample containers 202 held in the sample rack 201 are supplied to the sample analyzers 1 and 2 and the smear preparation device 3 by the transport device 4. This allows for the smooth supply of samples to each device as needed, enabling rapid analysis of samples and preparation of smears by the sample analyzers 1 and 2.

[0116] <Example of changes> In embodiments 1 to 3 described above, the diffractive optical element 114 may have a light-gathering effect. In this case, for example, the diffraction pattern formed on the diffractive optical element 114 may itself have a light-gathering effect, or a diffraction pattern that generates diffracted light may be formed on the incident surface of the diffractive optical element 114, and a pattern having a lens effect or a Fresnel lens may be formed on the outgoing surface of the diffractive optical element 114. If the diffractive optical element 114 has a light-gathering effect, the light-gathering lens 115 may be omitted.

[0117] In embodiments 1 to 3 described above, the diffractive optical element 114 was a transmissive diffractive optical element, but it may also be a reflective diffractive optical element.

[0118] In the embodiments 1 to 3 described above, the calculation unit 11 of the sample analyzer 1 classified cells using the AI ​​algorithm 32 based on the detection signals from the light receiving units 123, 133, and 143. However, the system is not limited to this, and cells may also be classified by comparing the patterns of the detection signals from the light receiving units 123, 133, and 143 with patterns previously stored in the storage unit 12.

[0119] Embodiments of the present invention can be modified in various ways as appropriate within the scope of the technical idea set forth in the claims. [Explanation of Symbols]

[0120] 1. Sample analyzer 10 Control Unit 21 Sample preparation section 100 Optical Systems 101 Flow Cell 111 Light source 114 Diffractive optical elements 115 Focusing lens 123, 133, 143 Light receiving section 124, 134, 144 dichroic mirrors (separated optical elements) 126, 136, 146 Light-receiving section (other light-receiving section) 151, 153 Light source (other light source) 157 Dichroic mirror (matching optical element) IS irradiation optical system

Claims

1. A light source that emits light, An illumination optical system that statically irradiates illumination light, which includes a diffractive optical element into which the light is incident, wherein multiple spots of diffracted light generated by the diffractive optical element are distributed in a predetermined pattern within the irradiation range, A flow cell through which a sample containing cells is flowed into the aforementioned irradiation area, The system includes a light-receiving unit that receives light generated from the cells flowing through the flow cell without separating it into diffracted light, based on the irradiation of the illumination light by the irradiation optical system, The illumination light includes zero-order diffracted light whose relative intensity to other diffracted light is 10 times or less. The irradiation optical system includes the spot of the zeroth order diffracted light among the multiple spots of diffracted light in the irradiation range. Optical system.

2. The illumination optical system focuses the illumination light generated from the diffractive optical element onto the flow cell. The relative intensity is the ratio of the amount of the zero-order diffracted light to the amount of the other diffracted light at the focusing surface of the illumination light. The optical system according to claim 1.

3. The amount of light from the other diffracted light is a representative value of the light intensity of at least some of the multiple spots formed by the other diffracted light. The optical system according to claim 2.

4. The aforementioned representative values ​​are the maximum value, mean, median, or mode. The optical system according to claim 3.

5. If L0 is the intensity of the incident light incident on the diffractive optical element, L1 is the intensity of the zero-order diffracted light, L2 is the intensity of other diffracted light besides the zero-order diffracted light, R1 is the ratio of the zero-order diffracted light generated from the diffractive optical element to the intensity of the incident light, R2 is the diffraction efficiency of the diffractive optical element, and N is the number of other diffracted light rays included in the illumination light, then The intensity L1 of the zero-order diffracted light is expressed by the following equation (1): The intensity L2 of the other diffracted light is expressed by the following equation (2): The aforementioned relative intensity is expressed by L1 / L2. The optical system according to claim 1. L1=L0×R1...(1) L2=(L0-L1)×R2 / N...(2)

6. The zero-order diffracted light has a relative intensity of 1 or less compared to other diffracted light contained in the illumination light. The optical system according to claim 1.

7. The ratio of the number of regions containing diffraction light other than the zero-order diffraction light to a plurality of grid-like regions in the illumination light, which include a region containing one of the diffraction lights and a region not containing the diffraction light, is 0.3% or more and 2% or less. The optical system according to claim 1.

8. The length of the illumination light in the flow direction of the sample in the flow cell is 300 pixels or more, where one pixel is defined as the region containing one of the diffracted light beams in the illumination light. The optical system according to claim 1.

9. The light-receiving unit receives scattered light generated from the cells flowing through the flow cell. The optical system according to claim 1.

10. The light-receiving unit receives fluorescence generated from the cells flowing through the flow cell. The optical system according to claim 1.

11. The illumination optical system further comprises matching optical elements that match other light emitted from other light sources to the illumination light, A separation optical element that separates the light generated from the cells by the illumination light from the light generated from the cells by the other light, The system further comprises another light-receiving unit that receives light based on the other light separated by the separating optical element, The optical system according to claim 1.

12. A sample preparation unit mixes a sample containing cells with reagents to prepare a sample, A light source that emits light, An illumination optical system that statically irradiates illumination light, which includes a diffractive optical element into which the light is incident, wherein multiple spots of diffracted light generated by the diffractive optical element are distributed in a predetermined pattern within the irradiation range, A flow cell through which a sample containing cells is flowed into the aforementioned irradiation area, A light receiving unit that receives light generated from the cells flowing through the flow cell by the irradiation of the illumination light by the irradiation optical system without separating it into diffracted light, The system includes a control unit that classifies the cells using an AI algorithm based on the signal from the light receiving unit, The illumination light includes zero-order diffracted light whose relative intensity to multiple other diffracted lights is 10 times or less. The irradiation optical system includes the spot of the zeroth order diffracted light among the multiple spots of diffracted light in the irradiation range. Sample analysis device.

13. The irradiation optical system includes a focusing lens that focuses the diffracted light generated from the diffracting optical element onto the flow cell. The specimen analyzer according to claim 12.

14. The light-receiving unit receives scattered light generated from the cells flowing through the flow cell. The specimen analyzer according to claim 12.

15. The light-receiving unit receives fluorescence generated from the cells flowing through the flow cell. The specimen analyzer according to claim 12.

16. The illumination optical system further comprises matching optical elements that match other light emitted from other light sources to the illumination light, A separation optical element that separates the light generated from the cells by the illumination light from the light generated from the cells by the other light, The system further comprises another light-receiving unit that receives light based on the other light separated by the separating optical element, The specimen analyzer according to claim 12.

17. The aforementioned specimen is one of the following: blood, cerebrospinal fluid, bone marrow fluid, or body cavity fluid. The specimen analyzer according to claim 12.

18. The aforementioned reagent contains a hemolytic agent. The specimen analyzer according to claim 12.

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