JP2008241644A - Particle analyzer - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a particle analyzer capable of determining precisely whether a measured signal is a particle signal, without complicating a process. <P>SOLUTION: A blood analyzer 1 is equipped with an FSC amplified waveform processing circuit 532, having an intermediate amplification factor for outputting a waveform signal acquired, by amplifying an electric signal outputted from a photodiode 406 with an amplification factor G1; an FSC amplified waveform processing circuit 542, having a high amplification factor for outputting a waveform signal acquired by amplifying the electrical signal outputted from the photodiode 406, with an amplification factor G2 which is different from the amplification factor G1; an operation part 651 for acquiring a peak value PB1 that shows particle characteristics, based on the waveform signal amplified with the amplification factor G1, and acquiring a peak value PC1 that shows particle characteristics, based on the waveform signal amplified with the amplification factor G2; and a control part 301 for determining whether the peak value PB1 and the peak value PC1 have a prescribed relation, and for acquiring analysis result. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a particle analyzer, and more particularly to a particle analyzer that analyzes particles by receiving light from the particles.

  2. Description of the Related Art Conventionally, a particle analyzer that analyzes particles by receiving light from the particles is known (see, for example, Patent Document 1).

  The flow cytometer (particle analyzer) described in Patent Document 1 includes a photoelectric conversion element that can optically detect the characteristics of each particle. The particle signal acquired by this photoelectric conversion element is represented by a pulse waveform having at least one peak, and in this pulse waveform, a peak value different from the particle signal is formed before and after the maximum peak value. There is. The flow cytometer described in Patent Document 1 is configured not to process a peak value different from a particle signal appearing before and after the maximum peak value as a particle signal. Specifically, this flow cytometer measures spherical standard particles having the same diameter as the particles to be measured as a preliminary step before the actual measurement step, thereby determining whether or not the actual signal is a particle signal. The peak interval necessary for making the determination and the ratio between the maximum peak value and the peak value different from the particle signal appearing before and after the maximum peak value are determined in advance. Then, based on the determined peak interval and the ratio of the maximum peak value and the peak value different from the particle signal appearing before and after the maximum peak value, whether or not the actually measured signal is a particle signal. It is configured to make such a determination.

JP 2001-281132 A

  However, in the flow cytometer (particle analyzer) described in Patent Document 1, when determining whether or not the actually measured signal is a particle signal, the peak interval, the maximum peak value, and the maximum peak value are determined. In order to predetermine the ratio of the peak value different from the signal of the particles appearing before and after, it is necessary to perform a preliminary process before the actual measurement process. This complicates the process for using the flow cytometer.

  The present invention has been made to solve the above-described problems, and one object of the present invention is to determine whether or not an actually measured signal is a particle signal without complicating the process. It is an object of the present invention to provide a particle analyzer capable of accurately performing the above.

Means for Solving the Problems and Effects of the Invention

  A particle analyzer according to one aspect of the present invention includes a flow cell that converts a particle-containing liquid containing particles into a flow of particles, an emission unit that emits light into the flow of particles, and an electrical signal that receives light from the particles. A light receiving unit that converts the electrical signal output from the light receiving unit with a first amplification factor, and a first signal amplification unit that outputs a first signal obtained by amplifying the electrical signal output from the light receiving unit. And a second signal amplifying means for outputting a second signal amplified with a different second amplification factor, a first characteristic parameter indicating the characteristics of the particle based on the first signal, and a particle signal based on the second signal Feature parameter acquisition means for acquiring a second feature parameter indicating a feature, determination means for determining whether or not the first feature parameter and the second feature parameter have a predetermined relationship, and the determination means has a predetermined relationship. Then judged And based on at least one of the first feature parameter and a second feature parameter and a analyzing unit for obtaining an analysis result.

  In the particle analyzer according to one aspect of the present invention, as described above, the first characteristic parameter acquired based on the first signal amplified with the first amplification factor and the second amplification factor different from the first amplification factor. Whether or not the actually measured signal is a particle signal is provided by providing a determination unit that determines whether or not the second characteristic parameter acquired based on the second signal amplified in step S2 has a predetermined relationship. Judgment can be made accurately. In addition, by providing an analysis unit that acquires an analysis result based on at least one of the first feature parameter and the second feature parameter determined to have a predetermined relationship by the determination unit, the actually measured signal is a particle signal. Since the particle analysis can be performed without the need for a preliminary process for determining whether or not the particle analysis apparatus is used, the process for using the particle analyzer is not complicated.

  In the particle analyzer according to the above aspect, the second amplification factor is preferably configured to be larger than the first amplification factor. If comprised in this way, using the 2nd feature parameter which shows the feature of the particle acquired based on the 2nd signal amplified with the 2nd amplification factor larger than the 1st amplification factor of the 1st signal, the 1st feature It can be determined whether the parameter and the second feature parameter have a predetermined relationship.

  In the particle analyzer according to the above aspect, preferably, the predetermined relationship is such that the coordinates represented by the first feature parameter and the second feature parameter are on the coordinates about the first feature parameter and the second feature parameter. It exists in the predetermined area. If comprised in this way, based on the 1st characteristic parameter acquired based on the 1st signal amplified with the 1st amplification factor and the 2nd signal amplified with the 2nd amplification factor different from the 1st amplification factor easily It is possible to determine whether or not the first feature parameter and the second feature parameter have a predetermined relationship by comparing the acquired second feature parameter.

  Particle analysis in which the predetermined relationship is a relationship in which the coordinates represented by the first feature parameter and the second feature parameter exist in a predetermined region on the coordinates with the first feature parameter and the second feature parameter as axes. In the apparatus, preferably, the predetermined region is a region including a straight line defined by the first amplification factor and the second amplification factor on the coordinates. With this configuration, it is possible to detect the first signal having the first characteristic parameter and the second signal having the second characteristic parameter corresponding to the relationship between the first amplification factor and the second amplification factor.

In the particle analyzer configured such that the predetermined region is located on the region side having a large inclination with respect to a straight line defined by the first amplification factor and the second amplification factor on the coordinates, The relationship is as follows when the peak value of the first signal is X, the peak value of the second signal is Y, the first gain is G1, the second gain is G2, and the coefficients are α and β: ).
Y ≦ α × G2 / G1 × X + β (1)
If comprised in this way, when the peak value X of the 1st signal amplified by 1st gain G1 is located in the area | region side which satisfy | fills Formula (1) on a coordinate, it will be amplified by 1st gain G1. Since the peak value X of the first signal can be regarded as a signal corresponding to the peak value Y of the second signal amplified by the second amplification factor G2 or a signal larger than the peak value Y. The peak value X of the first signal can be determined to be the peak value acquired from the particle signal.

  In the particle analyzer in which the predetermined relationship satisfies the expression (1), preferably, the first feature parameter and the second feature parameter are the peak value X of the first signal and the peak value Y of the second signal, respectively. The feature parameter acquisition means acquires the peak value of the pulse waveform that exceeds the first threshold value in the first signal as the peak value X of the first signal, and the second signal that is different from the first threshold value in the second signal. The peak value of the pulse waveform exceeding the threshold value is acquired as the peak value Y of the second signal, and the determination means is configured so that the peak value X of the first signal and the peak value Y of the second signal are expressed by the formula (1). It is configured to determine whether or not a predetermined relationship defined in the above is satisfied. According to this configuration, when the peak value of the pulse waveform exceeding the first threshold in the first signal does not indicate the maximum value of the first signal, the second threshold different from the first threshold in the second signal. The peak value of the pulse waveform exceeding the first threshold in the first signal is easily determined to be a peak value different from the peak value acquired by the particle signal, as compared with the peak value of the pulse waveform exceeding be able to.

In the particle analyzer configured so that the determination unit determines whether or not the peak value of the first signal and the peak value of the second signal satisfy the predetermined relationship defined in Equation (1), The determination unit is configured to exclude the peak value X of the first signal from the analysis result when the peak value X of the first signal and the peak value Y of the second signal satisfy the relationship of the following expression (2). Has been.
Y> α × G2 / G1 × X + β (2)
According to this configuration, the peak value different from the peak value acquired from the particle signal is excluded, so that the accuracy of particle analysis can be improved.

  In the particle analyzer including the discriminating unit that excludes the peak value of the first signal from the analysis result, preferably, the first signal included in the analysis result excluding the peak value X of the first signal that satisfies the relationship of Expression (2). First distribution map creating means for creating a first distribution map based on the signal is further provided. If comprised in this way, the particle | grain analysis result which excluded the peak value different from the peak value acquired by the signal of particle | grains can be provided with a 1st distribution map.

  In the particle analyzer according to the above aspect, it is preferable that the third signal that outputs the third signal obtained by amplifying the electric signal output from the light receiving unit with the third gain smaller than the first gain and the second gain. Amplifying means and second distribution map creating means for creating a second distribution map based on the third signal are further provided. With this configuration, the accurate peak value of the particles for which the first signal amplified with the first amplification factor and the second signal amplified with the second amplification factor are too large to obtain an accurate peak due to the amplification factor being too large can be obtained. 3 signals can be acquired, and the first signal amplified with the first amplification factor and the second signal amplified with the second amplification factor have an amplification factor that is too large to obtain an accurate peak. Particle analysis results can be provided by the second distribution map.

  In the particle analyzer according to the above aspect, the electrical signal is preferably a scattered light signal based on light scattered by the particles when the particles receive the light emitted by the emission unit. If comprised in this way, the optical signal reflecting the characteristic of particle | grains can be obtained.

  In the particle analyzer according to the above aspect, the particles preferably include red blood cells or platelets. If comprised in this way, the particle | grain analyzer which can acquire the analysis result of erythrocytes or platelets can be obtained.

  DESCRIPTION OF THE PREFERRED EMBODIMENTS Embodiments embodying the present invention will be described below with reference to the drawings.

  FIG. 1 is a perspective view showing the overall structure of a blood analyzer according to an embodiment of the present invention. FIG. 2 is a block diagram showing the configuration of the measurement unit and data processing diagram according to one embodiment of the present invention. 3-5 is a figure for demonstrating the structure of the blood analyzer by one Embodiment shown in FIG. First, with reference to FIGS. 1-5, the whole structure of the blood analyzer 1 by one Embodiment of this invention is demonstrated.

  The blood analyzer 1 according to one embodiment of the present invention guides particles such as cells and blood cells to the flow cell, irradiates each particle with laser light, and detects scattered light and fluorescence from each particle, thereby detecting blood cells in blood. It is a device that performs analysis.

  As shown in FIG. 1, the blood analyzer 1 includes a measurement unit 2 having a function of measuring blood as a sample, and a data processing unit that analyzes the measurement result output from the measurement unit 2 and obtains the analysis result. 3. As shown in FIG. 2, the measurement unit 2 includes a blood cell detection unit 4, an analog processing unit 5 for the output of the detection unit 4, a microcomputer unit 6, a display / operation unit 7, and a device for blood measurement. The mechanism part 8 is provided.

  In addition, as shown in FIG. 3, the measurement unit 2 is provided with a blood collection tube 11 filled with a predetermined amount of blood as a sample, a sampling valve 12 for sucking the blood sample, and a reaction chamber 13. Yes. The sampling valve 12 has a function of quantifying a blood sample sucked from a suction pipette (not shown) by a predetermined amount. The sampling valve 12 is configured to be able to mix a predetermined drug with a predetermined amount of blood sample. That is, the sampling valve 12 is configured to generate a diluted sample in which a predetermined drug is mixed with a predetermined amount of blood sample. The reaction chamber 13 is connected to the sampling valve 12 and is configured to further mix a predetermined drug with the diluted sample generated in the sampling valve 12. The reaction chamber 13 is connected to the detection unit 4, and is configured so that a measurement sample in which a predetermined drug is further mixed in the reaction chamber 13 flows into the detection unit 4.

  The detection unit 4 is configured as an optical detection unit. Specifically, the detection unit 4 is configured to detect cells such as cells and blood cells in blood by flow cytometry. Flow cytometry measures the physical and chemical properties of cells and other biological particles by passing them through narrow channels. It is a way for. The detection unit 4 is configured to detect forward scattered light, side scattered light, and side fluorescence emitted from blood cells in a sheath flow cell 403 irradiated with laser light to be described later. Here, light scattering is a phenomenon that occurs when particles such as blood cells exist as obstacles in the traveling direction of light and the light changes its traveling direction. By detecting this scattered light, it is possible to obtain information on the size and material of the particles. Specifically, it is possible to obtain information on the size of the particle (blood cell) by detecting forward scattered light, and obtain information on the inside of the particle by detecting side scattered light. Is possible. Side fluorescence is a phenomenon that occurs when a stained blood cell emits light when the blood cell stained with a fluorescent material is irradiated with laser light. Information on the degree of staining of blood cells can be obtained by detecting this side fluorescence.

  As shown in FIG. 4, the detection unit 4 includes a laser diode 401 that emits laser light, an irradiation lens unit 402, a sheath flow cell 403 through which the laser light passes, and a direction in which the laser light emitted from the laser diode 401 travels. , A condensing lens 404, a pinhole 405, and a photodiode 406 arranged on the extension line, a condensing lens 407 arranged in a direction crossing the direction in which the laser light emitted from the laser diode 401 travels, and a dichroic mirror 408, an optical filter 409, a pinhole 410 and a photodiode 411, and a photodiode 412 disposed on the side of the dichroic mirror 408.

  The laser diode 401 is provided to emit light to blood cells passing through the inside of the sheath flow cell 403. The irradiation lens unit 402 is configured to receive light emitted from the laser diode 401 and irradiate the sheath flow cell 403. As a result, the light emitted from the laser diode 401 is applied to the blood cells passing through the inside of the sheath flow cell 403. Then, forward scattered light, side scattered light, and side fluorescence are emitted by blood cells. The forward scattered light is emitted so as to travel on an extension line in the direction in which the laser light emitted from the laser diode 401 travels. The condensing lens 404 and the pinhole 405 arranged in the direction in which the forward scattered light travels have a function of condensing the forward scattered light and adjusting the focus of the forward scattered light. The photodiode 406 is provided to receive forward scattered light whose focus is adjusted by the condenser lens 404 and the pinhole 405.

  Side scattered light is emitted so as to travel in a direction intersecting with the traveling direction of the laser light emitted from the laser diode 401. The condensing lens 407 disposed in the direction in which the side scattered light travels is provided to collect the side scattered light. Further, the dichroic mirror 408 is configured to transmit the side scattered light collected by the condenser lens 407 to advance the side scattered light in the direction of the optical filter 409. The optical filter 409 and the pinhole 410 have a function of adjusting the focus of the side scattered light. The photodiode 411 is provided to receive side scattered light whose focus is adjusted by the pinhole 410.

  The side fluorescence is emitted so as to travel in a direction intersecting with the traveling direction of the laser light emitted from the laser diode 401. The condensing lens 407 arranged in the direction in which the side fluorescence advances has a function of condensing the side fluorescence together with the side scattered light. Further, the dichroic mirror 408 is configured to cause the side fluorescent light to advance to the photodiode 412 by reflecting the side fluorescent light collected by the condenser lens 407, unlike the side scattered light. The photodiode 412 is provided to receive the side fluorescence reflected by the dichroic mirror 408.

  In addition, each of the photodiode 406, the photodiode 411, and the photodiode 412 has a function of converting the received optical signal into an electrical signal. Further, as shown in FIG. 2, the detection unit 4 receives an electrical signal converted from an optical signal by the photodiode 406 (see FIG. 4), the photodiode 411 (see FIG. 4), and the photodiode 412 (see FIG. 4). It is connected so as to be transmitted to the analog processing unit 5.

  Here, in this embodiment, the analog processing unit 5 includes a side fluorescence detection circuit (SFL detection circuit) 501, a side scattered light detection circuit (SSC detection circuit) 511, and forward scattered light detection for a low amplification factor. A circuit (FSC detection circuit) 521, a forward scattered light detection circuit (FSC detection circuit) 531 for medium amplification factor, a forward scattered light detection circuit (FSC detection circuit) 541 for high amplification factor, and a side fluorescence amplification waveform Processing circuit (SFL amplification waveform processing circuit) 502, side scattered light amplification waveform processing circuit (SSC amplification waveform processing circuit) 512, low-amplification forward scattered light amplification waveform processing circuit (FSC amplification waveform processing circuit) 522, , A forward scattered light amplification waveform processing circuit (FSC amplification waveform processing circuit) 532 having a medium amplification factor, and a forward scattered light amplification waveform processing circuit (FSC amplification waveform processing circuit) 542 having a high amplification factor.

  The SFL detection circuit 501 has a function of detecting an electrical signal of side fluorescence transmitted from the photodiode 412 (see FIG. 4). The SFL amplification waveform processing circuit 502 is connected to the SFL detection circuit 501 and has a function of amplifying the side fluorescent electrical signal and processing the waveform of the amplified side fluorescent electrical signal. Further, the SSC detection circuit 511 has a function of detecting an electric signal of side scattered light transmitted from the photodiode 411 (see FIG. 4). The SSC amplification waveform processing circuit 512 is connected to the SSC detection circuit 511 and has a function of amplifying the side scattered light electrical signal and processing the waveform of the amplified side scattered light electrical signal.

  In this embodiment, the FSC detection circuit 521 for low amplification factor has a function of detecting an electrical signal of forward scattered light transmitted from the photodiode 406 (see FIG. 4). The FSC amplification waveform processing circuit 522 having a low amplification factor is connected to the FSC detection circuit 521 for the low amplification factor, and amplifies the forward scattered light by amplifying the electric signal of the forward scattered light at a predetermined amplification factor G0. It has a function of removing noise by waveform processing of the electrical signal. The FSC amplification waveform processing circuit 522 is provided for analyzing red blood cells and platelets passing through the inside of the sheath flow cell 403, and is particularly configured to analyze red blood cells having particles larger than platelets. ing.

  In the present embodiment, the FSC detection circuit 531 for medium amplification factor has a function of detecting an electrical signal of forward scattered light transmitted from the photodiode 406 (see FIG. 4). The medium gain FSC amplification waveform processing circuit 532 is connected to the medium gain FSC detection circuit 531, and amplifies the forward scattered light by amplifying the electric signal of the forward scattered light at a predetermined gain G1. It has a function of removing noise by waveform processing of the electrical signal. The amplification factor G1 is configured to be an amplification factor three times larger than the amplification factor G0. The medium amplification factor FSC amplification waveform processing circuit 532 is provided for analyzing red blood cells and platelets passing through the inside of the sheath flow cell 403, particularly for analyzing platelets having particles smaller than the red blood cells. Configured to suit.

  In the present embodiment, the FSC detection circuit 541 for high gain has a function of detecting an electrical signal of forward scattered light transmitted from the photodiode 406 (see FIG. 4). The high gain FSC amplification waveform processing circuit 542 is connected to the high gain FSC detection circuit 541, which amplifies the forward scattered light by amplifying the electric signal of the forward scattered light at a predetermined gain G2. It has a function of removing noise by waveform processing of the electrical signal. The amplification factor G2 is configured to be an amplification factor 10 times as large as the amplification factor G1. Further, the FSC amplification waveform processing circuit 542 having a high amplification factor, as will be described later, is an electrical signal (waveform signal) detected when the red blood cell R (see FIG. 4) passes through the sheath flow cell 403 (see FIG. 4). ) Is provided to distinguish the false signal (pseudo waveform signal) generated before and after the electrical signal (waveform signal) detected when platelets pass through the sheath flow cell 403. That is, the FSC amplification waveform processing circuit 542 having a high amplification factor is more detailed than the case where the control unit 301 described later analyzes the analysis of platelets having particles smaller than red blood cells only by the FSC amplification waveform processing circuit 532 having a medium amplification factor. It is provided for analysis.

  The analog processing unit 5 includes an SFL amplification waveform processing circuit 502, an SSC amplification waveform processing circuit 512, a low amplification factor FSC amplification waveform processing circuit 522, a medium amplification factor FSC amplification waveform processing circuit 532, and a high amplification factor FSC amplification. The electrical signal (waveform signal) subjected to waveform processing by the waveform processing circuit 542 is connected so as to be transmitted to the microcomputer unit 6.

  The microcomputer unit 6 includes a side fluorescence A / D conversion unit (SFLA / D conversion unit) 601, a side scattered light A / D conversion unit (SSCA / D conversion unit) 611, and a forward scattered light A for low amplification factor. / D conversion unit (FSCA / D conversion unit) 621, forward scattered light A / D conversion unit (FSCA / D conversion unit) 631 for medium amplification factor, and forward scattered light A / D conversion unit (FSCA) for high amplification factor / D conversion unit) 641, a calculation unit 651, an external connection interface 661, and a control unit 671.

  The SFLA / D converter 601, SSCA / D converter 611, FSCA / D converter 621 for low gain, FSCA / D converter 631 for medium gain, and FSCA / D converter 641 for high gain are SFL amplification waveform processing circuit 502, SSC amplification waveform processing circuit 512, low amplification factor FSC amplification waveform processing circuit 522, medium amplification factor FSC amplification waveform processing circuit 532, and high amplification factor FSC amplification waveform processing circuit 542, respectively. SFL amplification waveform processing circuit 502, SSC amplification waveform processing circuit 512, low amplification factor FSC amplification waveform processing circuit 522, medium amplification factor FSC amplification waveform processing circuit 532, and high amplification factor FSC amplification waveform processing. Each of the analog waveform signals of the circuit 542 has a function of converting into a digital waveform signal.

  In the present embodiment, the arithmetic unit 651 includes an SFLA / D conversion unit 601, an SSCA / D conversion unit 611, an FSCA / D conversion unit 621 for a low amplification factor, an FSCA / D conversion unit 631 for a medium amplification factor, and It is connected to the high gain FSCA / D converter 641 and has a function of executing a predetermined computer program on the digital waveform signal. The computing unit 651 has a function of acquiring a peak value of a digital waveform signal detected when particles (red blood cells and platelets) pass through the sheath flow cell 403 (see FIG. 4).

  The control unit 671 includes a control processor and a memory for operating the control processor. The control unit 671 has a function of controlling the apparatus mechanism unit 8 including a sampler (not shown) that automatically supplies the blood collection tube 11 (see FIG. 3), a fluid system for sample adjustment and measurement, and other controls. .

  The microcomputer unit 6 is provided with a bus 681 and a plurality of other interfaces. The characteristic parameters (peak values) of the digital waveform signal calculated by the calculation unit 651 are the interface 691, the bus 681, and the control unit. 671, the bus 682, and the external connection interface 661 are transmitted to the data processing unit 3. Further, the display / operation unit 7 and the device mechanism unit 8 for blood measurement are connected to the bus 681 via interfaces 692 and 693, respectively.

  As shown in FIG. 1, the data processing unit 3 (see FIG. 1) includes a personal computer (PC) or the like, and includes a control unit 301 including a CPU, ROM, RAM, and the like, a display unit 302, and an input device 303. Contains. The display unit 302 is provided for displaying an analysis result obtained by analyzing the digital signal data transmitted from the microcomputer unit 6.

  Next, the configuration of the data processing unit 3 will be described. As illustrated in FIG. 5, the data processing unit 3 is configured by a computer mainly configured by a control unit 301, a display unit 302, and an input device 303. The control unit 301 mainly includes a CPU 301a, a ROM 301b, a RAM 301c, a hard disk 301d, a reading device 301e, an input / output interface 301f, and an image output interface 301g. The CPU 301a, ROM 301b, RAM 301c, hard disk 301d, reading device 301e, input / output interface 301f, and image output interface 301g are connected by a bus 301h.

  The CPU 301a can execute computer programs stored in the ROM 301b and computer programs loaded in the RAM 301c. The computer functions as the data processing unit 3 when the CPU 301a executes an application program 304a described later.

  The ROM 301b is configured by a mask ROM, PROM, EPROM, EEPROM, or the like, and stores a computer program executed by the CPU 301a, data used for the same, and the like.

  The RAM 301c is configured by SRAM, DRAM, or the like. The RAM 301c is used to read out computer programs recorded in the ROM 301b and the hard disk 301d. Further, when these computer programs are executed, they are used as a work area of the CPU 301a.

  The hard disk 301d is installed with various computer programs to be executed by the CPU 301a, such as an operating system and application programs, and data used for executing the computer programs. An application program 304a described later is also installed in the hard disk 301d.

  The reading device 301e is configured by a flexible disk drive, a CD-ROM drive, a DVD-ROM drive, or the like, and can read a computer program or data recorded on the portable recording medium 304. The portable recording medium 304 stores an application program 304a for causing the computer to realize a predetermined function. The computer as the data processing unit 3 reads the application program 304a from the portable recording medium 304, and The application program 304a can be installed on the hard disk 301d.

  The application program 304a is not only provided by the portable recording medium 304, but also from an external device that is communicably connected to the data processing unit 3 through an electric communication line (whether wired or wireless). It can also be provided through a communication line. For example, the application program 304a is stored in a hard disk of a server computer on the Internet, and the data processing unit 3 accesses the server computer to download the application program 304a and install it on the hard disk 301d. Is also possible.

  In addition, an operating system that provides a graphical user interface environment, such as Windows (registered trademark) manufactured and sold by Microsoft Corporation, is installed in the hard disk 301d. In the following description, it is assumed that the application program 304a according to the present embodiment operates on the operating system.

  The input / output interface 301f includes, for example, a serial interface such as USB, IEEE 1394, and RS-232C, a parallel interface such as SCSI, IDE, and IEEE1284, and an analog interface including a D / A converter and an A / D converter. Has been. An input device 303 including a keyboard and a mouse is connected to the input / output interface 301f, and the user can input data to the data processing unit 3 by using the input device 303.

  The image output interface 301g is connected to a display unit 302 constituted by an LCD or a CRT, and outputs a video signal corresponding to the image data given from the CPU 301a to the display unit 302. The display unit 302 displays an image (screen) according to the input video signal.

  The application program 304a installed on the hard disk 301d of the control unit 301 of the data processing unit 3 uses the light emission amount (digital signal data) of the measurement sample transmitted from the microcomputer unit 6 of the measurement unit 2. The number of platelets using the peak values (feature parameters) of digital waveform signals acquired at different amplification factors (G1 and G2) in the calculation unit 651 during the RET measurement described later It is configured to analyze in detail.

  Specifically, the control unit 301 of the data processing unit 3 generates an electrical signal (waveform signal) detected when the red blood cell R (see FIG. 4) passes through the sheath flow cell 403 (see FIG. 4). The peak value acquired by the pseudo signal (pseudo waveform signal) generated before and after is different from the peak value acquired by the electrical signal (waveform signal) detected when platelets pass through the sheath flow cell 403. It has the function to determine that it has. Note that the peak value acquired by the false signal (pseudo waveform signal) generated before and after the electrical signal (waveform signal) detected when red blood cells pass through the sheath flow cell 403 is platelets inside the sheath flow cell 403. A method of determining that the signal has a characteristic different from the peak value acquired by the electric signal (waveform signal) detected when the signal passes will be described in detail later.

  The measurement unit 2 of the blood analyzer 1 according to the present embodiment has a function of measuring white blood cells and measuring red blood cells and platelets. Specifically, the measurement of leukocytes includes 4DIFF measurement, which measures fractions of lymphocytes, monocytes, eosinophils, neutrophils and basophils in leukocytes. In addition, the measurement of red blood cells and platelets includes NRBC measurement for fractional measurement of a population of nucleated red blood cells in blood cells and RET measurement for differential measurement of a population of reticulocytes and platelets in blood cells. The data processing unit 3 of the blood analyzer 1 has a function of analyzing data measured in 4DIFF measurement, NRBC measurement, and RET measurement.

  6 to 8 are diagrams showing waveform signals processed based on the forward scattered light when the RET measurement is performed using the blood analyzer according to the embodiment shown in FIG. FIG. 9 is a plane for determining whether or not the peak value acquired when the RET measurement is performed using the blood analyzer according to the embodiment shown in FIG. 1 is the peak value acquired by the false signal. It is the figure which showed the coordinate. Next, referring to FIGS. 2, 4, and 6 to 9, a pseudo signal (pseudo waveform) generated before and after an electrical signal (waveform signal) detected when red blood cells pass through the sheath flow cell 403. The method for determining that the peak value acquired by the signal) has a different characteristic from the peak value acquired by the electrical signal (waveform signal) detected when the platelets pass through the sheath flow cell 403. explain.

  First, as shown in FIG. 4, the forward scattered light when the red blood cells R pass through the sheath flow cell 403 of the detection unit 4 is detected by the photodiode 406, and the optical signal of the forward scattered light is an analog electrical signal. Converted to a signal. Then, the electrical signal converted based on the optical signal of the forward scattered light is processed into a waveform signal A, a waveform signal B, and a waveform signal C of a digital signal through a flow described later.

  As shown in FIG. 6, the waveform signal A is processed by being amplified at an amplification factor G0 suitable for detecting red blood cells by a low amplification factor FSC amplification waveform processing circuit 522 (see FIG. 2). Further, the calculation unit 651 (see FIG. 2) determines that the skirt portions A1 and A2 on both sides of the mountain-shaped waveform portion of the waveform signal A are based on the threshold reference value A0 having a predetermined signal intensity. The peak value PA1 of the signal portion A3 that crosses the signal is acquired as a signal of the red blood cell R (see FIG. 4). The reference threshold A0 is set in advance so as to have a signal strength greater than the peak values PA2 and PA3 of the false signals A4 and A5 appearing before and after the signal portion A3 of the waveform signal A.

  Further, in the present embodiment, the waveform signal B (pulse waveform signal) is suitable for detecting platelets by a medium amplification factor FSC amplification waveform processing circuit 532 (see FIG. 2), as shown in FIG. Further, it is processed by being amplified with an amplification factor G1 that is three times as large as the amplification factor G0. In addition, the calculation unit 651 (see FIG. 2) applies a reference threshold value B0 having a predetermined signal intensity to the waveform signal processed by the FSC amplification waveform processing circuit 532 having a medium amplification factor, thereby generating particles (red blood cells and platelets). ) Is passed through the inside of the sheath flow cell 403 (see FIG. 4). However, when the red blood cell R (see FIG. 4) passes through the inside of the sheath flow cell 403, the reference threshold value B0 having a predetermined signal intensity is equal to the skirt portions B1 on both sides of the mountain-shaped waveform portion of the waveform signal B and Cross B2. In this case, the calculation unit 651 acquires the peak value PB1 of the signal portion B3, which is a false signal generated when the red blood cell R passes through the sheath flow cell 403, as a particle signal. For this reason, in the FSC amplification waveform processing circuit 532 (see FIG. 2) having the medium amplification factor, the peak value PB2 of the signal portion B4 that should be originally acquired is not acquired.

  Further, in the present embodiment, as shown in FIG. 8, the waveform signal C is amplified at a gain G2 that is ten times larger than the gain G1 by the FSC amplification waveform processing circuit 542 (see FIG. 2) having a high gain. It is processed by being amplified. In addition, the FSC amplification waveform processing circuit 542 having a high amplification factor is a pseudo signal (for example, a waveform signal B) acquired by the waveform signal (for example, the waveform signal B) processed by the FSC amplification waveform processing circuit 532 (see FIG. 2) having a medium amplification factor. For example, it is provided to process the peak value (for example, peak value PB1) of the signal portion B3) so as not to be a particle signal. Specifically, the calculation unit 651 (see FIG. 2) uses the reference threshold value C0 having a predetermined signal intensity as a reference for the skirt portions C1 and C2 on both sides of the mountain-shaped waveform portion of the waveform signal C. The peak value PC1 of the signal portion C3 straddling the threshold value C0 is obtained as a signal of the red blood cell R (see FIG. 4). The reference threshold value C0 is determined from the valley portion PC2 between the false signal C4 and the signal portion C3 appearing before and after the signal portion C3 of the waveform signal C, and the valley portion PC3 between the signal portion C3 and the false signal C5. Is also preset to have a small signal strength.

  Here, in this embodiment, as shown in FIGS. 7 and 8, the control unit 301 (see FIG. 2) of the data processing unit 3 is configured such that the skirt portions on both sides of the mountain-shaped waveform portion of the waveform signal B. The peak value PB1 of the signal portion B3 where B1 and B2 straddle the reference threshold value B0, and the peak value of the signal portion C3 where the skirt portions C1 and C2 on both sides of the mountain-shaped waveform portion of the waveform signal C straddle the reference threshold value C0. Comparing with PC1, the peak value PB1 is determined to be a peak value acquired by a false signal generated when the red blood cell R passes through the sheath flow cell 403. Specifically, the control unit 301 of the data processing unit 3 determines that the peak value PB1 acquired at the amplification factor G1 is 10 minutes of the peak value PC1 acquired at the amplification factor G2 having an amplification factor 10 times the amplification factor G1. It is determined whether or not it substantially matches 1.

In the present embodiment, as shown in FIG. 9, the control unit 301 (see FIG. 1) of the data processing unit 3 determines whether or not the peak value PB1 substantially matches one-tenth of the peak value PC1. In order to do this, a plane coordinate is configured with the X-axis as the forward scattered light intensity acquired at the amplification factor G1 and the Y-axis as the forward scattered light intensity acquired at the amplification factor G2. In addition, in this plane coordinate, the peak value of the waveform signal acquired at the amplification factor G1 is X, the peak value of the waveform signal acquired at the amplification factor G2 is Y, and the coefficients are α (α> 0) and β. A region that satisfies the following equation (4) based on the equation (3) (Y = 10 × X (3)) of a straight line having an inclination of 10 (G2 / G1 = 10) Area) is configured.
Y> α × 10 × X + β (4)
That is, the region is configured on the side where the inclination is large with respect to the linear expression (Y = α × 10 × X + β) of Expression (4). In addition, α and β in the equation (4) are on the side where the slope is larger than the linear equation (Y = α × 10 × X + β) in the equation (4) in consideration of measurement errors of the peak values X and Y, etc. Has a function of giving a width to the straight line (Y = 10 × X) of the formula (3) serving as a reference of the region of FIG. 3 to determine whether the peak value Y is 10 times the peak value X or not. . Then, the peak value X (for example, the peak value PB1) of the waveform signal acquired at the amplification factor G1 corresponding to the particles (for example, the red blood cell R) that has passed through the inside of the sheath flow cell 403 in a region that satisfies the relationship of the expression (4). When the plot with the peak value Y (for example, peak value PC1) of the waveform signal acquired at the amplification factor G2 is located, the control unit 301 determines that the peak value Y does not substantially correspond to 10 times the peak value X. The peak value (for example, peak value PB1) of the waveform signal acquired at the amplification factor G1 is determined to be the peak value of the false signal. On the other hand, the waveform acquired with the peak value X (for example, peak value P1) and the gain G2 of the waveform signal acquired with the amplification factor G1 corresponding to the particles that passed through the inside of the sheath flow cell 403 in the region not satisfying the equation (4). When a plot with the peak value Y (for example, peak value P2) of the signal is located, the control unit 301 determines that the peak value Y is substantially equivalent to 10 times the peak value X, and obtains the waveform obtained with the amplification factor G1. The signal peak value X (for example, PB2) is determined not to be a peak value due to a false signal. That is, when the following expression (5) is satisfied, the control unit 301 determines that the peak value Y is substantially equivalent to 10 times the peak value X, and the peak value X (for example, the peak value X of the waveform signal acquired at the amplification factor G1) It is determined that PB2) is not a peak value due to a false signal.
Y ≦ α × 10 × X + β (5)

  FIG. 10 is a flowchart for explaining the measurement and processing flow of the measurement unit and the data processing unit of the blood analyzer according to the present embodiment shown in FIG. FIGS. 11 to 13 and FIG. 15 are diagrams showing scattergrams representing the results measured and analyzed by the blood analyzer according to the present embodiment shown in FIG. Next, referring to FIG. 1 to FIG. 5, FIG. 10 to FIG. 13 and FIG. 15, when blood analyzer 1 according to one embodiment of the present invention performs and analyzes 4DIFF measurement, NRBC measurement, and RET measurement. The operation will be described. First, the operation of the measurement unit 2 will be described in detail.

  In step S1 of FIG. 10, when a main switch (not shown) of the measurement unit 2 (see FIG. 1) is turned on, the control unit 671 is initialized and the operation of each unit of the measurement unit 2 is checked. When a main switch (not shown) of the data processing unit 3 (see FIG. 1) is turned on, the control unit 301 of the data processing unit 3 is initialized (program initialization) in step S21.

  Next, in step S22, a menu screen (not shown) is displayed on the display unit 302 (see FIG. 1), and the measurement mode of any of 4DIFF measurement, NRBC measurement, and RET measurement displayed on the menu screen by the user is displayed. Is selected, it is determined whether or not the measurement mode has been input (input has been accepted). If it is determined in step S22 that the measurement mode has been input, the process proceeds to step S23. In step S23, the measurement mode is changed to any one of 4DIFF measurement, NRBC measurement, and RET measurement selected by the user, and the process proceeds to step S24. If it is determined in step S22 that the measurement mode is not input, the process proceeds to step S24.

  Thereafter, in step S24, the CPU 301a (see FIG. 5) determines whether or not a measurement start instruction instructed by the user has been received. If it is determined in step S24 that a measurement start instruction has not been received, the process proceeds to step S25. If it is determined in step S24 that a measurement start instruction has been received, the process proceeds to step S26. In step S26, a measurement start signal including measurement mode information is transmitted to the control unit 671 via the external connection interface 661 of the microcomputer unit 6 (see FIG. 2) of the measurement unit 2.

  Next, in step S2, the measurement start signal transmitted by the user operating the data processing unit 3 (see FIG. 2) is received by the control unit 671 (see FIG. 2) of the microcomputer unit 6 of the measurement unit 2. It is determined whether or not. If it is determined in step S2 that the measurement start signal has not been received, the process proceeds to step S3. If it is determined in step S2 that the measurement start signal has been received, the process proceeds to step S4.

  Thereafter, in step S4, the control unit 671 stores any one of the 4DIFF measurement, NRBC measurement, and RET measurement measurement modes input in the data processing unit 3. In step S5, a blood sample in the blood collection tube 11 (see FIG. 3) is sucked into a sampling valve 12 (see FIG. 3) from a suction pipette (not shown), and a predetermined amount of blood sample is a predetermined amount of drug (hemolysis). Diluted with an agent or diluent). In step S5, the blood sample diluted with the drug (haemolytic agent or diluent) is transferred to the reaction chamber 13 (see FIG. 3).

  Next, in step S6, a predetermined amount of drug (staining solution) is supplied to the blood sample diluted with the drug (hemolyzing agent or diluent), the diluted sample is further diluted, and desired blood cells are stained. In this state, the measurement sample is obtained by reacting the diluted sample within the reaction chamber 13 for a predetermined time. In step S6, the measurement sample is sent to the detection unit 4 (see FIG. 3) together with the sheath liquid, and the process proceeds to step S7.

  Thereafter, in step S7, the control unit 671 determines whether or not the 4DIFF measurement mode has been set by the user operating the data processing unit 3. When it is determined in step S7 that the 4DIFF measurement mode is set, the process proceeds to step S8. In step S8, the measurement sample is sent to the detection unit 4 together with the sheath liquid, whereby 4DIFF measurement is performed, and the process proceeds to step S9. The operation of 4DIFF measurement will be described later in detail. If it is determined in step S7 that the 4DIFF measurement mode is not set, the process proceeds to step S9.

  Thereafter, in step S9, the control unit 671 determines whether or not the RET measurement mode is set by the user operating the data processing unit 3. If it is determined in step S9 that the RET measurement mode is set, the process proceeds to step S10. In step S10, RET measurement is performed, and the process proceeds to step S11. The operation of RET measurement will be described later in detail. If it is determined in step S9 that the RET measurement mode is not set, the process proceeds to step S11.

  Thereafter, in step S11, the control unit 671 determines whether or not the user has set the NRBC measurement mode by operating the data processing unit 3. If it is determined in step S11 that the NRBC measurement mode is set, the process proceeds to step S12. In step S12, NRBC measurement is performed, and the process proceeds to step S13. The operation of NRBC measurement will be described later in detail. If it is determined in step S11 that the NRBC measurement mode is not set, the process proceeds to step S13.

  Next, in step S13, 4DIFF measurement, RET measurement, and NRBC measurement data are transmitted to the data processing unit 3 via the external connection interface 661, and the process proceeds to step S27.

  Thereafter, in step S27, the measurement unit 2 measures the 4DIFF measurement data, the NRBC measurement data, and the RET measurement data that are measured by the measurement unit 2 and then transmitted through the external connection interface 661 (see FIG. 2). It is determined whether any data is received via the input / output interface 301f (see FIG. 5). In step S27, when it is determined that the data measured by the measurement unit 2 among the 4DIFF measurement data, the NRBC measurement data, and the RET measurement data is not received, the 4DIFF measurement data, the NRBC measurement data, and the RET This determination is repeated until data measured by the measurement unit 2 is received. If it is determined in step S27 that the data measured by the measurement unit 2 among the 4DIFF measurement data, the NRBC measurement data, and the RET measurement data is received, the process proceeds to step S28.

  Next, in step S28, the data measured by the measurement unit 2 among the 4DIFF measurement data, the NRBC measurement data, and the RET measurement data received in step S27 by the ROM 301b (see FIG. 5) and the RAM 301c (see FIG. 5). Remember.

  Thereafter, in step S29, the CPU 301a determines whether the 4DIFF measurement data is stored in the ROM 301b and the RAM 301c. If it is determined in step S29 that 4DIFF measurement data is stored, the process proceeds to step S30. In step S30, 4DIFF measurement data is processed, and the process proceeds to step S31. The operation of processing 4DIFF measurement data will be described in detail later. If it is determined in step S29 that 4DIFF measurement data is not stored, the process proceeds to step S31.

  Thereafter, in step S31, the CPU 301a determines whether the RET measurement data is stored in the ROM 301b and the RAM 301c. If it is determined in step S31 that RET measurement data is stored, the process proceeds to step S32. In step S32, RET measurement data is processed, and the process proceeds to step S33. The operation of processing the RET measurement data will be described later in detail. If it is determined in step S31 that RET measurement data is not stored, the process proceeds to step S33.

  Thereafter, in step S33, the CPU 301a determines whether the NRBC measurement data is stored in the ROM 301b and the RAM 301c. If it is determined in step S33 that NRBC measurement data is stored, the process proceeds to step S34. In step S34, the NRBC measurement data is processed, and the process proceeds to step S35. The operation of processing the NRBC measurement data will be described in detail later. If it is determined in step S33 that NRBC measurement data is not stored, the process proceeds to step S35.

  Thereafter, in step S35, the 4DIFF measurement DIFF scattergram (see FIG. 11) created in step S30, the RET measurement RET scattergram (red blood cell scattergram) (see FIG. 12) and PLT created in step S32. At least one of the scattergram (platelet scattergram) (see FIG. 13) and the NRBC scattergram (see FIG. 15) of the NRBC measurement created in step S34 is displayed on the display unit 302 (FIG. 1). Displayed).

  Next, in step S25, the CPU 301a determines whether or not a shutdown instruction has been issued (whether or not a shutdown button (not shown) has been pressed from the menu screen), and it is determined that a shutdown instruction has been issued. If it is determined that the shutdown instruction has not been issued, the process returns to step S22. In step S36, a shutdown signal is transmitted from the CPU 301a to the control unit 671 of the microcomputer unit 6 of the measurement unit 2, and the process ends.

  Next, in step S3, it is determined whether or not the shutdown signal transmitted from the data processing unit 3 has been received. If it is determined that the shutdown signal has been received, the process proceeds to step S14 and measurement is performed. The unit 2 is shut down, and the process ends. If it is determined in step S3 that the shutdown signal has not been received, the process returns to step S2.

  FIG. 16 is a flowchart for explaining details of the 4DIFF measurement executed in step S8 of the flowchart shown in FIG. Next, a 4DIFF measurement flow executed in the measurement unit 2 of the blood analyzer 1 according to the present embodiment will be described with reference to FIGS. 2 to 4, 10, and 16.

  First, in step S41 of FIG. 16, the control unit 671 (see FIG. 2) reads the setting values of the 4DIFF measurement mode in order to perform 4DIFF measurement. In step S42, the control unit 671 sets signal processing circuits (SFL detection circuit 501 and SSC detection circuit 511) (see FIG. 2) used in 4DIFF measurement based on the 4DIFF measurement mode.

  Next, in step S43, the measurement sample moved to the detection unit 4 (see FIG. 3) is circulated inside the sheath flow cell 403 (see FIG. 4), whereby the supply of the measurement sample is started. In step S44, the laser diode 401 (see FIG. 4) oscillates the laser to start measuring the blood cells of the measurement sample. Thereby, by irradiating the blood cells circulating in the sheath flow cell 403 with laser light, forward scattered light, side scattered light, and side fluorescence are emitted from the blood cells. The side scattered light and the side fluorescence emitted from the blood cells are received by the photodiodes 411 and 412 and converted into analog electric signals, respectively. Then, the electric signal of the side scattered light and the electric signal of the side fluorescence are transmitted to the SSC detection circuit 511 and the SFL detection circuit 501 respectively.

  Thereafter, in step S45, the electrical signal of the side scattered light and the electrical signal of the side fluorescence are transmitted to the SSC amplification waveform processing circuit 512 and the SFL amplification waveform processing circuit 502, respectively, and are amplified with a desired amplification factor. Waveform processing is performed. In step S46, the analog waveform signal of the side scattered light and the analog waveform signal of the side fluorescence are transmitted to the SSCA / D conversion unit 611 and the SFLA / D conversion unit 601, respectively, and an analog waveform is obtained. The signal is converted into a digital waveform signal.

  In step S47, a peak value is acquired from the digital signal of side scattered light and the waveform signal of side fluorescence by the calculation unit 651 (see FIG. 2), and the acquired peak value is the interface 691 (FIG. 2). 2), bus 681 (see FIG. 2), control unit 671 (see FIG. 2), bus 682 (see FIG. 2), and external connection interface 661 (see FIG. 2) to the data processing unit 3 (see FIG. 2). Sent.

  Next, in step S48, the control unit 671 determines whether or not a predetermined time has elapsed for the measurement time of the blood cells of the measurement sample. If it is determined in step S48 that the time measured for the blood cells of the measurement sample has not elapsed, the process returns to step S45. That is, the operations of Steps S45 to S48 are repeated until a predetermined time elapses for the measurement sample blood cells. If it is determined in step S48 that the time measured for the blood cells of the measurement sample has elapsed, the process proceeds to step S49 and the supply of the measurement sample is stopped.

  Thereafter, in step S50, the device mechanism unit 8 is operated by a command from the control unit 671, and the sheath flow cell 403 is washed.

  FIG. 17 is a flowchart for explaining details of the RET measurement executed in step S10 of the flowchart shown in FIG. Next, the RET measurement flow executed in the measurement unit 2 of the blood analyzer 1 according to the present embodiment will be described with reference to FIGS. 2 to 4, 6 to 8, 10 and 17.

  First, in step S51 of FIG. 17, the control unit 671 (see FIG. 2) reads the set value of the RET measurement mode and the like in order to perform RET measurement. In step S52, the control unit 671 uses the signal processing circuit (SFL detection circuit 501, low amplification factor FSC detection circuit 521, medium amplification factor FSC detection circuit) used in the RET measurement based on the RET measurement mode. 531 and the FSC detection circuit 541 for high gain (see FIG. 2) are set.

  Next, in step S53, the measurement sample moved to the detection unit 4 (see FIG. 3) is circulated in the sheath flow cell 403 (see FIG. 4), whereby supply of the measurement sample is started. In step S54, the laser diode 401 (see FIG. 4) oscillates the laser to start measuring the blood cells of the measurement sample. Thereby, by irradiating the blood cells circulating in the sheath flow cell 403 with laser light, forward scattered light, side scattered light, and side fluorescence are emitted from the blood cells. The forward scattered light and the side fluorescence emitted from the blood cells are received by the photodiodes 406 and 412 and converted into analog electric signals, respectively. The electrical signal of the forward scattered light is branched into three signals, and the FSC detection circuit 521 for low amplification factor, the FSC detection circuit 531 for medium amplification factor, and the FSC detection circuit 541 for high amplification factor, respectively. Sent to. Further, the electrical signal of the side fluorescence is transmitted to the SFL detection circuit 501.

  Thereafter, in step S55, the three electric signals branched from the forward scattered light are converted into an FSC amplification waveform processing circuit 522 having a low amplification factor, an FSC amplification waveform processing circuit 532 having a medium amplification factor, and an FSC amplification waveform processing circuit having a high amplification factor. 542. Then, it is amplified to an amplification factor G0 by the FSC amplification waveform processing circuit 522 having a low amplification factor and subjected to waveform processing. Further, the signal is amplified to the amplification factor G1 by the medium amplification factor FSC amplification waveform processing circuit 532 and processed, and the waveform is processed by the amplification factor G2 by the high amplification factor FSC amplification waveform processing circuit 542. . In addition, the side fluorescence electric signal is transmitted to the SFL amplification waveform processing circuit 502, and is amplified with a desired amplification factor to be subjected to waveform processing. In step S56, the analog waveform signals of the forward scattered light amplified at the amplification factor G0, the amplification factor G1, and the amplification factor G2 are converted into the low amplification factor FSCA / D conversion unit 621 and the medium amplification factor, respectively. The signal is transmitted to the FSCA / D conversion unit 631 and the FSCA / D conversion unit 641 for high gain, and is converted from an analog waveform signal to a digital waveform signal. Further, the side waveform analog waveform signal is transmitted to the SFLA / D converter 601 and is converted from an analog waveform signal to a digital waveform signal.

  In step S57, for example, peak values PA1 and PB1 are respectively obtained from the waveform signals of the digital signals of the forward scattered light amplified by the calculation unit 651 (see FIG. 2) with the amplification factor G0, the amplification factor G1, and the amplification factor G2. And PC1 (see FIGS. 6, 7 and 8) are acquired. For example, the acquired peak values PA1, PB1 and PC1 are the interface 691 (see FIG. 2), the bus 681 (see FIG. 2), and the control unit 671 (see FIG. 2), the bus 682 (see FIG. 2), and the external connection interface 661 (see FIG. 2), the data is transmitted to the data processing unit 3 (see FIG. 2). Further, a peak value is acquired from the waveform signal of the side fluorescence, and the acquired peak value is transmitted to the data processing unit 3 via the interface 691, the bus 681, the control unit 671, the bus 682, and the external connection interface 661. The

  Next, in step S58, the control unit 671 determines whether or not a predetermined time has elapsed for the measurement time of the blood cells of the measurement sample. If it is determined in step S58 that the time measured for the blood cells of the measurement sample has not elapsed, the process returns to step S55. That is, the operations of Step S55 to Step S58 are repeated until a predetermined time elapses for the measurement sample blood cells. If it is determined in step S58 that the time measured for the blood cells of the measurement sample has elapsed, the process proceeds to step S59, and the supply of the measurement sample is stopped.

  Thereafter, in step S60, the device mechanism unit 8 is operated by a command from the control unit 671, and the sheath flow cell 403 is washed.

  FIG. 18 is a flowchart for explaining details of the NRBC measurement executed in step S12 of the flowchart shown in FIG. Next, an NRBC measurement flow executed in the measurement unit 2 of the blood analyzer 1 according to the present embodiment will be described with reference to FIGS.

  First, in step S61 of FIG. 18, the control unit 671 (see FIG. 2) reads the set value of the NRBC measurement mode and the like in order to perform NRBC measurement. In step S62, the control unit 671 sets the signal processing circuit (SFL detection circuit 501 and low gain FSC detection circuit 521) (see FIG. 2) used in the NRBC measurement based on the NRBC measurement mode. Done.

  Next, in step S63, the measurement sample moved to the detection unit 4 (see FIG. 3) is circulated inside the sheath flow cell 403 (see FIG. 4), whereby the supply of the measurement sample is started. In step S64, the laser diode 401 (see FIG. 4) oscillates the laser to start measuring the blood cells of the measurement sample. Thereby, by irradiating the blood cells circulating in the sheath flow cell 403 with laser light, forward scattered light, side scattered light, and side fluorescence are emitted from the blood cells. The forward scattered light and the side fluorescence emitted from the blood cells are received by the photodiodes 406 and 412 and converted into analog electric signals, respectively. Then, the electrical signal of the forward scattered light is transmitted to the FSC detection circuit 521 for low amplification factor. Further, the electrical signal of the side fluorescence is transmitted to the SFL detection circuit 501.

  Thereafter, in step S65, the electrical signal of the forward scattered light is transmitted to the FSC amplification waveform processing circuit 522 having a low amplification factor. Then, it is amplified to an amplification factor G0 by the FSC amplification waveform processing circuit 522 having a low amplification factor and subjected to waveform processing. In addition, the side fluorescence electric signal is transmitted to the SFL amplification waveform processing circuit 502, and is amplified with a desired amplification factor to be subjected to waveform processing. In step S66, the analog waveform signal of the forward scattered light is transmitted to the low amplification factor FSCA / D conversion unit 621, and converted from the analog waveform signal to a digital waveform signal. Further, the side waveform analog waveform signal is transmitted to the SFLA / D converter 601 and is converted from an analog waveform signal to a digital waveform signal.

  In step S67, the calculation unit 651 (see FIG. 2) acquires a peak value from the waveform signal of the digital signal of the forward scattered light, and the acquired peak value is the interface 691 (see FIG. 2) and the bus 681. (See FIG. 2), the data is transmitted to the data processing unit 3 (see FIG. 2) via the control unit 671 (see FIG. 2), the bus 682 (see FIG. 2), and the external connection interface 661 (see FIG. 2). Further, a peak value is acquired from the waveform signal of the side fluorescence, and the acquired peak value is transmitted to the data processing unit 3 via the interface 691, the bus 681, the control unit 671, the bus 682, and the external connection interface 661. The

  Next, in step S68, the control unit 671 determines whether or not a predetermined time has elapsed for the measurement sample blood cells. If it is determined in step S68 that the time measured for the blood cells of the measurement sample has not elapsed, the process returns to step S65. That is, the operations in steps S65 to S68 are repeated until the time measured for the blood cells of the measurement sample elapses a predetermined time. In step S68, if it is determined that the time measured for the blood cells of the measurement sample has elapsed, the process proceeds to step S69, and the supply of the measurement sample is stopped.

  Thereafter, in step S70, the device mechanism unit 8 is operated by a command from the control unit 671, and the sheath flow cell 403 is washed.

  FIG. 19 is a flowchart for explaining details of the 4DIFF measurement data processing executed in step S30 of the flowchart shown in FIG. Next, a 4DIFF measurement data processing flow executed in the data processing unit 3 of the blood analyzer 1 according to the present embodiment will be described with reference to FIGS. 1, 5, 10, 11 and 19.

  First, in step S71 of FIG. 19, a plurality of peak values acquired by the control unit 301 (see FIGS. 1 and 5) based on the waveform signal of side scattered light and the waveform signal of side fluorescence are measured. Read from. In step S72, a plurality of sets of plots of a peak value based on the waveform signal of the side scattered light detected at the same time and a peak value based on the waveform signal of the side fluorescence are horizontally plotted by the control unit 301. A scattergram (see FIG. 11) of 4DIFF measurement expressed on coordinates having the side fluorescent intensity on the axis and the side scattered light intensity on the vertical axis is created.

  FIG. 14 is a diagram showing a scattergram for explaining the scattergram shown in FIG. 13 in detail. FIG. 20 is a flowchart for explaining details of the RET measurement data processing executed in step S32 of the flowchart shown in FIG. Next, referring to FIGS. 1, 5, 9, 10, 12-14 and 20, the RET measurement data processing flow executed in the data processing unit 3 of the blood analyzer 1 according to the present embodiment. Will be explained.

  First, in step S81 of FIG. 20, a plurality of peak values PA1, PB1, and PC1 acquired by the control unit 301 (see FIGS. 1 and 5) based on the waveform signal of forward scattered light and the waveform signal of side fluorescence. Are read from the measurement unit 2 and stored in the ROM 301b (see FIG. 5). In step S82, based on the peak value and the waveform signal of the side fluorescence acquired by the CPU 301a based on the waveform signal of the forward scattered light amplified at the amplification factors G0, G1, and G2 detected at the same time. One set is selected from a plurality of sets with the acquired peak value.

  Then, in step S83, the plot (X, Y) of each peak value acquired by the CPU 301a based on the waveform signal of the forward scattered light amplified at the amplification factors G1 and G2 detected at the same time is expressed by the equation (X). 5) It is determined whether or not Y ≦ α × 10 × X + β is satisfied (whether or not the region (see FIG. 9) satisfying Expression (4) (Y> α × 10 × X + β) is determined). ). Then, in step S83, a plot (X, Y) of each peak value acquired based on the waveform signal of the forward scattered light amplified at the amplification factors G1 and G2 detected at the same time is expressed by the equation (5) Y If it is determined that ≦ α × 10 × X + β is not satisfied (when it is determined that the region (see FIG. 9) that satisfies Expression (4) (Y> α × 10 × X + β) is present), the process proceeds to step S84. move on. In step S84, the peak value acquired based on the waveform signal of the forward scattered light amplified at the amplification factor G1 is deleted from the ROM 301b, and the process proceeds to step S85. Further, in step S83, the plot (X, Y) of each peak value acquired based on the waveform signal of the forward scattered light amplified at the amplification factors G1 and G2 detected at the same time is expressed by the equation (5) Y If it is determined that ≦ α × 10 × X + β is satisfied (when it is determined that the region does not exist in the region satisfying the formula (4) (Y> α × G2 / G1 × X + β) (see FIG. 9)), Proceed to S85.

  Thereafter, in step S85, based on the peak value and the waveform signal of the side fluorescence acquired by the CPU 301a based on the waveform signal of the forward scattered light amplified at the amplification factors G0, G1, and G2 detected at the same time. It is determined whether or not the determination in step S83 has been made for all of the plurality of pairs with the acquired peak value. And in step S85, it acquired based on the peak value acquired based on the waveform signal of the forward scattered light amplified by the amplification factors G0, G1, and G2 detected at the same time and the waveform signal of the side fluorescence. If it is determined that the determination in step S83 has not been made for all of the plurality of pairs with the peak value, the process proceeds to step S86. In step S86, the peak value acquired by the CPU 301a based on the waveform signal of the forward scattered light amplified at the amplification factors G0, G1, and G2 detected at the same time when the determination in step S83 is not performed. A pair with the peak value acquired based on the waveform signal of the side fluorescence is selected, and the process returns to step S83. That is, the peak value acquired based on the waveform signal of the forward scattered light amplified by the amplification factors G0, G1 and G2 detected at all the same time and the peak value acquired based on the waveform signal of the side fluorescence Steps S83 to S85 are repeated until the determination in step S83 is made for the pair. Moreover, in step S85, it acquired based on the peak value acquired based on the waveform signal of the forward scattered light amplified by the amplification factors G0, G1, and G2 detected at the same time and the waveform signal of the side fluorescence. If it is determined that the determination in step S83 has been made for all of the plurality of pairs with the peak value, the process proceeds to step S87.

  Thereafter, in step S87, the control unit 301 acquires the peak value acquired based on the waveform signal of the forward scattered light amplified by the amplification factor G0 and the amplification factor G1, and the peak signal acquired based on the waveform signal of the side fluorescence. The value is read. In step S88, the control unit 301 acquires the peak value acquired based on the waveform signal of the forward scattered light amplified at the amplification factor G0 detected at the same time and the waveform signal of the side fluorescence. A scattergram (see FIG. 12) of the RET measurement is generated in which a plurality of sets of plots with the peak values are represented on the coordinates having the lateral fluorescence intensity on the horizontal axis and the forward scattered light intensity on the vertical axis. Next, in step S89, the control unit 301 acquires the peak value acquired based on the waveform signal of the forward scattered light amplified at the amplification factor G1 detected at the same time and the waveform signal of the side fluorescence. A scattergram of PLT measurement (see FIG. 13) is created, in which a plurality of sets of plots with the peak values obtained are represented on coordinates with the lateral fluorescence intensity on the horizontal axis and the forward scattered light intensity on the vertical axis. It should be noted that the scattergram of the PLT measurement displayed on the display unit 302 was determined not to satisfy the formula (5) Y ≦ α × 10 × X + β in the above-described step S84 (formula (4) (Y> α × 10 (Plot of peak values acquired based on waveform signals of forward scattered light amplified at amplification rates G1 and G2 detected at the same time) (determined to exist in a region satisfying (XX + β)) (see FIG. 9) Since X, Y) is deleted, the plot of the area Z of the scattergram in FIG. 14 is not displayed. Further, the horizontal axis of the scattergram of PLT measurement is displayed logarithmically.

  FIG. 21 is a flowchart for explaining details of the NRBC measurement data processing executed in step S34 of the flowchart shown in FIG. Next, the NRBC measurement data processing flow executed in the data processing unit 3 of the blood analyzer 1 according to the present embodiment will be described with reference to FIGS. 1, 5, 10, 15, and 21. FIG.

  First, in step S91 in FIG. 21, a plurality of peak values acquired based on forward scattered light and side fluorescence are read from the measurement unit 2 by the control unit 301 (see FIGS. 1 and 5). In step S92, a plurality of sets of plots of the peak value based on the forward scattered light and the peak value based on the side fluorescence detected at the same time by the control unit 301 are plotted on the horizontal axis. A scattergram (see FIG. 15) of the NRBC measurement expressed on the coordinate having the forward scattered light intensity on the vertical axis is created.

  In the present embodiment, as described above, the peak value PB1 acquired based on the waveform signal B amplified with the amplification factor G1 and the waveform signal C amplified with the amplification factor G2 different from the amplification factor G1 are acquired. By providing determination means for determining whether or not the peak value PC1 has a predetermined relationship, it is possible to accurately determine whether or not the actually measured signal is a particle signal. Further, by providing the control unit 301 that acquires the analysis result based on the peak value PB1 and the peak value PC1 determined to have a predetermined relationship by the control unit 301, it is determined whether or not the actually measured signal is a particle signal. Since the particle analysis can be performed without the need for a preliminary process for making this determination, the process for using the particle analyzer can be simplified.

In the present embodiment, as described above, the predetermined relationship for determining whether or not the actually measured signal is a particle signal is X, and the peak value of the waveform signal amplified by the amplification factor G1 is X and amplified. When the peak value of the waveform signal amplified at an amplification factor G2 which is an amplification factor 10 times the rate G1 is Y and the coefficients are α (α> 0) and β, a relationship satisfying the following expression (5) is established. By
Y ≦ α × 10 × X + β (5)
When the peak value X of the waveform signal amplified by the amplification factor G1 is located on the region side satisfying the equation (5) on the coordinates, the peak value X of the waveform signal amplified by the amplification factor G1 is changed to the amplification factor G2. Since the signal corresponds to the peak value Y of the waveform signal amplified by the above, or can be regarded as a signal larger than the peak value Y, the peak value X of the waveform signal is obtained from the particle signal. It can be determined that it is a value.

  Further, in the present embodiment, as described above, the control unit 301 determines that the peak value of the waveform signal amplified by the amplification factor G1 and the peak value of the waveform signal amplified by the amplification factor G2 are expressed by the equation (4) (Y > Α × G2 / G1 × X + β), the peak acquired from the particle signal is configured by excluding the peak value of the waveform signal amplified by the amplification factor G1 from the analysis result. Since the peak value different from the value (the peak value acquired by the false signal) is excluded, the accuracy of the particle analysis can be improved.

  In the present embodiment, as described above, the waveform amplified by the amplification factor G1 in a state where the peak value of the waveform signal amplified by the amplification factor G1 existing in the region of the expression (4) is excluded from the analysis result. Particle analysis result excluding the peak value (peak value acquired by false signal) different from the peak value acquired by the particle signal by providing the control unit 301 that creates the scattergram of PLT measurement based on the signal Can be provided by the scattergram of the PLT measurement.

  In the present embodiment, as described above, a low-amplification FSC amplification waveform that outputs a waveform signal obtained by amplifying the electrical signal output from the photodiode 406 with an amplification factor G0 smaller than the amplification factor G1 and the amplification factor G2. By providing the processing circuit 522 and the control unit 301 that creates a scattergram of the RET measurement based on the waveform signal amplified by the amplification factor G0, the waveform signal amplified by the amplification factor G1 and the waveform signal amplified by the amplification factor G2 Then, an accurate peak value of red blood cells for which an amplification factor is too large to obtain an accurate peak value can be obtained from a waveform signal amplified at an amplification factor G0, and a waveform signal and an amplification factor amplified at an amplification factor G1 can be obtained. The waveform signal amplified by G2 has an amplification factor that is too large to obtain an accurate peak value. The constant of the scattergram, it is possible to provide.

  The embodiment disclosed this time should be considered as illustrative in all points and not restrictive. The scope of the present invention is shown not by the above description of the embodiment but by the scope of claims for patent, and includes all modifications within the meaning and scope equivalent to the scope of claims for patent.

  For example, in the above-described embodiment, an example in which the present invention is applied to a blood analyzer that is an example of an analyzer has been described. However, the present invention is not limited thereto, and may be applied to other analyzers such as a urine analyzer. Good.

  Moreover, in the said embodiment, the peak acquired by the false signal (fake waveform signal) which generate | occur | produces before and after the waveform signal detected when a red blood cell passes the inside of a sheath flow cell using the control part of a data processing part. Although an example is shown in which it is determined that the value (feature parameter) is determined, the present invention is not limited to this, and the detection is performed when red blood cells pass through the inside of the sheath flow cell using the control unit of the measurement unit. The peak value (feature parameter) acquired by the false signal (pseudo waveform signal) generated before and after the waveform signal may be determined.

  Moreover, although the example which provided the measurement part and the data processing part separately was shown in the said embodiment, this invention is not limited to this, You may provide a measurement part and a data processing part integrally.

  In the above embodiment, an example in which the present invention is mainly applied to the RET measurement for measuring red blood cells and platelets has been shown. However, the present invention is not limited to this, for example, white blood cells such as 4DIFF measurement, and other blood cells. The present invention may be applied to the measurement.

1 is a perspective view showing an overall structure of a blood analyzer according to an embodiment of the present invention. It is a block diagram which shows the structure of the measurement part and data processing figure by one Embodiment of this invention. It is a figure for demonstrating the structure of the detection part of the blood analyzer by one Embodiment shown in FIG. It is a figure for demonstrating the structure of the detection part and analog part of the blood analyzer by one Embodiment shown in FIG. It is a figure for demonstrating the structure of the data processing part of the blood analyzer by one Embodiment shown in FIG. It is the figure which showed the waveform signal processed based on the forward scattered light at the time of performing RET measurement using the blood analyzer by one Embodiment shown in FIG. It is the figure which showed the waveform signal processed based on the forward scattered light at the time of performing RET measurement using the blood analyzer by one Embodiment shown in FIG. It is the figure which showed the waveform signal processed based on the forward scattered light at the time of performing RET measurement using the blood analyzer by one Embodiment shown in FIG. A plane for determining whether or not the peak value (feature parameter) acquired when the RET measurement is performed using the blood analyzer according to the embodiment shown in FIG. 1 is the peak value acquired by a false signal. It is the figure which showed the coordinate. It is a flowchart for demonstrating the measurement and processing flow of the measurement part of a blood analyzer by this embodiment shown in FIG. 1, and a data processing part. It is the figure which showed the scattergram showing the measurement of 4DIFF measurement and the result analyzed by the blood analyzer by this embodiment shown in FIG. It is the figure which showed the scattergram of RET showing the measurement of RET measurement by the blood analyzer by this embodiment shown in FIG. 1, and the result analyzed. It is the figure which showed the scattergram of PLT showing the measurement of RET measurement by the blood analyzer by this embodiment shown in FIG. 1, and the result analyzed. It is the figure which showed the scattergram for demonstrating in detail the scattergram shown in FIG. It is the figure which showed the scattergram showing the measurement of NRBC measurement by the blood analyzer by this embodiment shown in FIG. 1, and the result analyzed. It is a flowchart for demonstrating the detail of 4DIFF measurement performed in the flowchart shown in FIG. It is a flowchart for demonstrating the detail of the RET measurement performed in the flowchart shown in FIG. 11 is a flowchart for explaining details of NRBC measurement executed in the flowchart shown in FIG. 10. It is a flowchart for demonstrating the detail of 4DIFF measurement data processing performed in the flowchart shown in FIG. It is a flowchart for demonstrating the detail of the RET measurement data process performed in the flowchart shown in FIG. FIG. 11 is a flowchart for explaining details of NRBC measurement data processing executed in the flowchart shown in FIG. 10. FIG.

Explanation of symbols

1 Blood analyzer (particle analyzer)
301 control unit (determination means, analysis means, first distribution chart creation means, second distribution chart creation means)
401 Laser diode (emitter)
403 Sheath flow cell (flow cell)
406 Photodiode (light receiving part)
522 Forward scattered light (FSC) amplification waveform processing circuit (third signal amplification means)
532 Forward scattered light (FSC) amplification waveform processing circuit (first signal amplification means)
542 Forward scattered light (FSC) amplification waveform processing circuit (second signal amplification means)
651 calculation unit (feature parameter acquisition means)
B0 reference threshold (first threshold)
B1 Bottom part B2 Bottom part C0 Reference threshold (second threshold)
C1 Bottom part C2 Bottom part G0 Gain (third gain)
G1 gain (first gain)
G2 gain (second gain)
PB1 peak value (first characteristic parameter)
PC1 peak value (second feature parameter)

Claims (11)

A flow cell for converting a particle-containing liquid containing particles into a flow of particles;
An emission part for emitting light to the flow of particles;
A light receiving unit that receives light from the particles and converts the light into an electrical signal;
First signal amplifying means for outputting a first signal obtained by amplifying the electric signal output from the light receiving unit with a first amplification factor;
Second signal amplification means for outputting a second signal obtained by amplifying the electrical signal output from the light receiving unit with a second amplification factor different from the first amplification factor;
Feature parameter acquisition means for acquiring a first feature parameter indicating the feature of the particle based on the first signal and acquiring a second feature parameter indicating the feature of the particle based on the second signal;
Determining means for determining whether or not the first feature parameter and the second feature parameter have a predetermined relationship;
A particle analysis apparatus comprising: an analysis unit that acquires an analysis result based on at least one of the first feature parameter and the second feature parameter determined to have the predetermined relationship by the determination unit.   The particle analyzer according to claim 1, wherein the second amplification factor is configured to be larger than the first amplification factor.   The predetermined relationship is that the coordinates represented by the first feature parameter and the second feature parameter exist in a predetermined region on the coordinates with the first feature parameter and the second feature parameter as axes. The particle analyzer according to claim 1 or 2, wherein the relationship is a relationship.   The particle analysis apparatus according to claim 3, wherein the predetermined region is a region including a straight line defined by the first amplification factor and the second amplification factor on the coordinates. The predetermined relationship is such that the peak value of the first signal is X, the peak value of the second signal is Y, the first gain is G1, the second gain is G2, and the coefficients are α and β. The particle analyzer according to claim 4, wherein the particle analyzer is configured to satisfy the following formula (1).
Y ≦ α × G2 / G1 × X + β (1) The first characteristic parameter and the second characteristic parameter are a peak value X of the first signal and a peak value Y of the second signal, respectively.
The characteristic parameter acquisition means includes
The peak value of the pulse waveform exceeding the first threshold in the first signal is acquired as the peak value X of the first signal, and the pulse waveform exceeding the second threshold different from the first threshold in the second signal is acquired. It is configured to acquire the peak value as the peak value Y of the second signal,
The determination unit is configured to determine whether or not a peak value X of the first signal and a peak value Y of the second signal satisfy a predetermined relationship defined in the equation (1). Item 6. The particle analyzer according to Item 5. The determination means determines the peak value X of the first signal from the analysis result when the peak value X of the first signal and the peak value Y of the second signal satisfy the relationship of the following expression (2). The particle analyzer according to claim 6, wherein the particle analyzer is configured to be excluded.
Y> α × G2 / G1 × X + β (2)   A first distribution map creating means for creating a first distribution map based on the first signal included in the analysis result excluding the peak value X of the first signal that satisfies the relationship of the formula (2); The particle analyzer according to claim 7. Third signal amplification means for outputting a third signal obtained by amplifying the electrical signal output from the light receiving unit with a third amplification factor smaller than the first amplification factor and the second amplification factor;
The particle analyzer according to any one of claims 1 to 8, further comprising second distribution map creating means for creating a second distribution map based on the third signal.   The particle according to any one of claims 1 to 9, wherein the electrical signal is a scattered light signal based on light scattered by the particle when the particle receives light emitted from the emission unit. Analysis equipment.   The particle analysis apparatus according to claim 1, wherein the particles include red blood cells or platelets.

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