JP6958166B2 - Analytical equipment and analytical method - Google Patents

Description

The present invention relates to an analyzer and an analytical method for analyzing biological substances such as antigens and antibodies.

An immunoassay (immunoassay) is known in which a specific antigen or antibody associated with a disease is detected as a biomarker to quantitatively analyze the effects of detection and treatment of the disease.

In Patent Document 1, an antibody fixed in a reaction region on a sample analysis disk and an antigen in a sample are bound, the antigen is labeled with fine particles having the antibody, and laser light emitted from an optical pickup is scanned. Thereby, an analyzer for detecting the fine particles captured in the reaction region is described. The analyzer described in Patent Document 1 uses an optical disk device for sample detection.

Japanese Unexamined Patent Publication No. 2015-127691

In a conventional analyzer as described in Patent Document 1, a cartridge is attached to a sample analysis disc to form a well. A reaction region is formed by injecting a sample solution and a buffer solution into the wells and advancing the antigen-antibody reaction. That is, the well functions as a container for storing the sample solution and the buffer solution.

However, air bubbles may adhere to the bottom surface of the well when the sample solution and the buffer solution are injected. When bubbles adhere to the surface of the sample analysis disk constituting the bottom surface of the well, the antigen-antibody reaction does not proceed in the region to which the bubbles adhere (hereinafter referred to as the bubble region). Therefore, a reaction region including a bubble region is formed on the sample analysis disc. Therefore, in the reaction region including the bubble region, it is difficult to accurately measure the fine particles in the reaction region.

An object of the present invention is to provide an analyzer and an analysis method capable of identifying a bubble region included in a reaction region and correcting the measurement result of fine particles in the bubble region.

The present invention includes a turntable for holding a sample analysis disk having a reaction region in which fine particles bound to a substance to be detected are captured for each track, a turntable drive unit for rotating the turntable, and the turntable. The turntable drive circuit that controls the drive unit and the turntable are driven along the direction orthogonal to the rotation axis of the turntable, irradiate the reaction region with laser light, receive the reflected light from the reaction region, and receive the light receiving level. An optical pickup that generates a signal, an optical pickup drive circuit that controls the drive of the optical pickup, and a control unit that controls the turntable drive circuit and the optical pickup drive circuit are provided, and the control unit is the reaction region. A plurality of measurement gate signals for measuring the number of the fine particles captured in the light signal are continuously generated for each track, the number of the fine particles is measured for each measurement gate signal from the light receiving level signal, and the reaction is performed. comparing the number of each of the particles measured before each Symbol measurement gate signal which is in a symmetrical position relationship with respect to the dividing line that passes through the center of the area, the measurement result correction target region for correcting the number of the fine particles providing analysis device that identifies the.

Further, in the present invention, a sample analysis disk having a reaction region in which fine particles bound to a substance to be detected are captured for each track is rotated, the reaction region is irradiated with laser light for each track, and the reaction region is radiated from the reaction region. A light receiving level signal is generated by receiving the reflected light of the above, and a plurality of measurement gate signals for measuring the number of the fine particles captured in the reaction region are continuously generated for each track, and the light receiving level signal is generated. the number of the fine particles for each of the measurement gate signal measured from, comparing the number of each of the fine particles of the measured in each measurement gate signal which is in a symmetrical position relationship with respect to the dividing line passing through the center of the reaction region and to provide a minute析方method that identifies the measurement result correction target region for correcting the number of the fine particles.

According to the analyzer and the analysis method of the present invention, the bubble region included in the reaction region can be specified, and the measurement result of the fine particles in the bubble region can be corrected.

It is a top view which shows the detection target substance capture unit. It is sectional drawing of the detection target substance capture unit cut in AA of FIG. It is sectional drawing which shows the state which the cartridge was removed from the disk for sample analysis. It is an enlarged perspective view which shows the well cut by BB of FIG. It is a flowchart for demonstrating the method of forming a reaction region on a disk for sample analysis. It is sectional drawing which shows the state which injected the buffer solution containing an antibody into a well in the process of forming a reaction region on a disk for sample analysis. It is sectional drawing which shows the state which injected the sample liquid containing the substance to be detected into a well in the process of forming a reaction region on a disk for sample analysis. It is sectional drawing which shows the state which injected the buffer solution containing fine particles into a well in the process of forming a reaction region on a disk for sample analysis. It is a schematic cross-sectional view which shows the state in which the substance to be detected is sandwiched and captured in the recess of the track region by an antibody and fine particles. It is a schematic plan view which shows the state which the fine particle is trapped in the concave part of the track area in the state which is bound with the substance to be detected. It is a block diagram which shows the analyzer of 1st and 2nd Embodiment. It is a top view for demonstrating the positional relationship between the detection position of a reference position detection sensor and an optical pickup, a notch part of a sample analysis disk, and a reaction area. It is a flowchart for demonstrating the method of analyzing fine particles by the analyzer of 1st and 2nd Embodiment. It is a time chart which shows the relationship between the reaction region and the measurement gate signal in the method of analyzing fine particles by the analyzer of 1st Embodiment. It is a flowchart for demonstrating the method of specifying a bubble region by the analyzer of 1st and 2nd Embodiment, and the method of correcting the measurement result of the fine particle in a bubble region. It is a time chart which shows the relationship between the adjacent track and the measurement gate signal in the method of analyzing fine particles by the analyzer of 1st Embodiment. It is a time chart which shows the relationship between the reaction region and the measurement gate signal in the method of analyzing fine particles by the analyzer of 2nd Embodiment. It is a time chart which shows the relationship between the adjacent track and the measurement gate signal in the method of analyzing fine particles by the analyzer of 2nd Embodiment.

[Detection target substance capture unit]
An example of the substance capture unit to be detected will be described with reference to FIGS. 1 to 3. FIG. 1 shows a state in which the substance capture unit to be detected is viewed from the cartridge side. FIG. 2A shows a cross section of the substance capture unit to be detected cut in AA of FIG. FIG. 2B shows a state in which the cartridge is removed from the sample analysis disc. FIG. 3 shows a partially enlarged state in which the well of FIG. 1 is cut by BB.

As shown in FIG. 1, the substance capture unit 100 to be detected includes a sample analysis disc 200 and a cartridge 300. The sample analysis disc 200 has a disk shape equivalent to that of an optical disc such as a Blu-ray disc (BD), a DVD, or a compact disc (CD). The sample analysis disc 200 is made of, for example, a resin material such as a polycarbonate resin or a cycloolefin polymer generally used for optical discs. The sample analysis disc 200 is not limited to the above-mentioned optical discs, and optical discs conforming to other forms or predetermined standards can also be used.

As shown in FIG. 1, FIG. 2A, or FIG. 2B, the sample analysis disc 200 has a central hole 201 formed in the central portion and a notch portion 202 formed in the outer peripheral portion. The notch 202 is a reference position identification unit for identifying the reference position of the sample analysis disc 200.

As shown in FIG. 3, on the surface of the sample analysis disc 200, a track region 205 in which convex portions 203 and concave portions 204 are alternately arranged in the radial direction is formed. The convex portion 203 and the concave portion 204 are formed in a spiral shape from the inner peripheral portion to the outer peripheral portion. The convex portion 203 corresponds to the land of the optical disc. The recess 204 corresponds to the groove of an optical disc. The track pitch corresponding to the radial pitch of the recess 204 is, for example, 320 nm.

As shown in FIGS. 1, 2A, or 2B, the cartridge 300 is formed with a plurality of cylindrical through holes 301 in the circumferential direction. The plurality of through holes 301 are formed at equal intervals so that their centers are located on the same circumference. The cartridge 300 has a convex portion 302 formed in the central portion and a convex portion 303 formed in the outer peripheral portion.

When the cartridge 300 is attached to the sample analysis disc 200, the convex portion 302 is inserted into the center hole 201 of the sample analysis disc 200, and the convex portion 303 is inserted into the notch 202, whereby the cartridge 300 and the sample analysis are used. The disk 200 can be positioned.

As shown in FIGS. 2A and 3, the substance capture unit 100 to be detected has a plurality of wells 101 formed by the through hole 301 of the cartridge 300 and the track region 205 of the sample analysis disc 200. The inner peripheral surface of the through hole 301 constitutes the inner peripheral surface of the well 101, and the track region 205 of the sample analysis disc 200 constitutes the bottom surface of the well 101. The well 101 is a container for storing a solution such as a sample solution and a buffer solution. Although FIG. 1 shows eight wells 101 as an example, the number of wells 101 is not limited to this.

As shown in FIG. 2B, the cartridge 300 can be separated from the sample analysis disc 200. The detection and measurement of the fine particles labeling the substance to be detected are performed by the sample analysis disc 200 alone from which the cartridge 300 is separated.

[Formation of reaction region]
An example of a method of forming a reaction region on the sample analysis disk 200 of the substance capture unit 100 to be detected will be described with reference to the flowchart of FIG. 4, FIG. 5A, FIG. 5B, FIG. 5C, FIG. 6, and FIG.

In step S101, the operator injects the buffer solution 112 containing the antibody 111 into the well 101 of the substance capture unit 100 to be detected, as shown in FIG. 5A. The operator incubates the substance capture unit 100 to be detected for an appropriate time and at an appropriate temperature. As a result, the antibody 111 is fixed on the track region 205 of the sample analysis disc 200 that constitutes the bottom surface of the well 101.

When the buffer solution 112 is injected, bubbles 113 may adhere to the track region 205 of the sample analysis disc 200 constituting the bottom surface of the well 101. The bubbles 113 tend to adhere to the boundary between the inner peripheral surface and the bottom surface of the well 101, that is, the boundary between the inner peripheral surface of the through hole 301 of the cartridge 300 and the track region 205 of the sample analysis disc 200. When the bubble 113 is attached on the track region 205, the antibody 111 is not fixed on the track region 205 in the bubble region to which the bubble 113 is attached.

The operator drains the buffer 112 from the well 101 and cleans the well 101 with another buffer. Antibody 111 that was not immobilized in the track region 205 is removed by this washing.

In step S102, the operator injects the sample liquid 122 containing the substance to be detected 121 into the well 101 as shown in FIG. 5B. The substance to be detected 121 is, for example, an exosome. The sample liquid 122 may not contain the substance to be detected 121. In order to make the explanation easy to understand, the case where the sample liquid 122 contains the substance to be detected 121 will be described.

The operator incubates the substance capture unit 100 to be detected for an appropriate time and at an appropriate temperature. As a result, the substance to be detected 121 specifically binds to the antibody 111 immobilized on the track region 205 by an antigen-antibody reaction. As a result, the substance to be detected 121 is captured in the track region 205.

When the sample liquid 122 is injected, bubbles 123 may adhere to the track region 205 of the sample analysis disc 200 constituting the bottom surface of the well 101. Similarly, the bubble 123 tends to adhere to the boundary between the inner surface and the bottom surface of the well 101, that is, the boundary between the inner peripheral surface of the through hole 301 of the cartridge 300 and the track region 205 of the sample analysis disc 200.

When the bubble 123 adheres to the track region 205, the detection target substance 121 cannot bind to the antibody 111 immobilized on the track region 205 in the bubble region to which the bubble 123 adheres. Therefore, in the bubble region, the substance 121 to be detected is not captured on the track region 205.

When the bubble region is formed in the track region 205 in step S101, the antibody 111 that binds to the detection target substance 121 is not fixed on the track region 205 in the bubble region. Therefore, even if the bubble 123 does not adhere to the track region 205 when the sample liquid 122 is injected, the detection target substance 121 is not captured on the track region 205 in the bubble region formed in step S101.

The operator drains the sample solution 122 from the well 101 and cleans the well 101 with a buffer solution. The detection target substance 121 dispersed in the sample solution 122 without binding to the antibody 111 and the detection target substance 121 adhering to the track region 205 by non-specific adsorption that is not an antigen-antibody reaction are washed. Is removed by.

In step S103, the operator injects the buffer solution 132 containing the labeling fine particles 131 into the well 101, as shown in FIG. 5C. An antibody that specifically binds to the substance to be detected 121 by an antigen-antibody reaction is formed on the surface of the fine particles 131.

The operator incubates the substance capture unit 100 to be detected for an appropriate time and at an appropriate temperature. As a result, the fine particles 131 specifically bind to the substance to be detected 121 captured on the track region 205 by an antigen-antibody reaction. As a result, the fine particles 131 are captured in the track region 205, specifically, the recess 204 of the track region 205 in a state of being bound to the detection target substance 121.

When the buffer solution 132 is injected, bubbles 133 may adhere to the track region 205 of the sample analysis disc 200 constituting the bottom surface of the well 101. Similarly, the bubble 133 tends to adhere to the boundary between the inner surface and the bottom surface of the well 101, that is, the boundary between the inner peripheral surface of the through hole 301 of the cartridge 300 and the track region 205 of the sample analysis disc 200. When the bubble 133 adheres to the track region 205, the fine particle 131 cannot bind to the detection target substance 121 captured on the track region 205 in the bubble region to which the bubble 133 adheres. Therefore, in the bubble region, the fine particles 131 are not captured on the track region 205.

When the bubble region is formed in the track region 205 in step S101 or step S102, the detection target substance 121 that binds to the fine particles 131 is not fixed on the track region 205 in the bubble region. Therefore, even if the bubbles 133 do not adhere to the track region 205 when the buffer solution 132 is injected, the fine particles 131 are not fixed on the track region 205 in the bubble region formed in step S101 or step S102.

The operator drains the buffer 132 from the well 101, cleans the well 101 with another buffer, and dries it. The fine particles 131 dispersed in the buffer solution 132 without binding to the detection target substance 121 are removed by washing.

In step S104, the operator separates the cartridge 300 of the substance capture unit 100 to be detected and the sample analysis disk 200, as shown in FIG. 2B. The sample analysis disc 200 is formed with a plurality of circular reaction regions 210 corresponding to the plurality of wells 101.

As shown in FIG. 6, in the reaction region 210, the fine particles 131 are captured in the recess 204 of the track region 205 in a state of being bound to the substance to be detected 121. That is, the substance to be detected 121 is sandwiched and captured in the recess 204 of the track region 205 by the antibody 111 and the fine particles 131. FIG. 7 shows an example of a state in which the fine particles 131 are captured in the recess 204 of the track region 205 in a state of being bound to the substance to be detected 121.

[First Embodiment]
The analyzer and the analysis method of the first embodiment will be described with reference to FIGS. 8 to 13. The analyzer of the first embodiment will be described with reference to FIG. When the substance 121 to be detected is an exosome, the size of the exosome is as small as about 100 nm, so that it is difficult to directly detect it optically. The analyzer 1 of the first embodiment indirectly detects the detection target substance 121 specifically bound to the fine particles 131 by detecting and measuring the fine particles 131 captured in the reaction region 210. measure.

The analyzer 1 includes a turntable 2, a clamper 3, a turntable drive unit 4, a turntable drive circuit 5, and a reference position detection sensor 6. Further, the analyzer 1 includes a guide shaft 7, an optical pickup 20, an optical pickup drive circuit 8, a control unit 9, a storage unit 10, and a display unit 11. The analyzer 1 does not have to include the display unit 11, and an external display unit may be used.

A sample analysis disc 200 is placed on the turntable 2 so that the reaction region 210 faces downward. The clamper 3 is driven in the direction of separation and approach to the turntable 2, that is, in the upward and downward directions of FIG. The sample analysis disc 200 is held by the clamper 3 and the turntable 2 when the clamper 3 is driven downward.

The turntable drive unit 4 rotationally drives the turntable 2 together with the sample analysis disc 200 and the clamper 3 around the rotation axis C2. A spindle motor may be used as the turntable drive unit 4. The turntable drive circuit 5 controls the turntable drive unit 4. For example, the turntable drive circuit 5 controls the turntable drive unit 4 so that the turntable 2 rotates at a constant linear velocity together with the sample analysis disc 200 and the clamper 3.

The reference position detection sensor 6 is arranged near the outer peripheral portion of the sample analysis disc 200. The reference position detection sensor 6 is an optical sensor such as a photoreflector. The reference position detection sensor 6 irradiates the outer peripheral portion of the sample analysis disc 200 with the detection light 6a while the sample analysis disc 200 is rotating, and receives the reflected light from the sample analysis disc 200.

The reference position detection sensor 6 detects the notch 202 of the sample analysis disk 200, generates the reference position detection signal KS, and outputs the reference position detection signal KS to the control unit 9. The reference position detection signal KS is a pulse signal that rises and turns on when the notch 202 reaches the detection position of the reference position detection sensor 6, that is, the position where the detection light 6a is irradiated, and falls and turns off when the notch 202 passes through. Is.

That is, the reference position detection sensor 6 detects the reference position for each rotation cycle and track of the sample analysis disc 200. A transmissive optical sensor may be used as the reference position detection sensor 6. In this case, the reference position detection sensor 6 irradiates the sample analysis disk 200 with the detection light 6a and receives the detection light 6a passing through the notch 202, whereby the rotation cycle of the sample analysis disk 200 and each track. Detects the reference position.

The guide shaft 7 is arranged parallel to the sample analysis disc 200 and along the radial direction of the sample analysis disc 200. The optical pickup 20 is supported by the guide shaft 7. The optical pickup 20 is driven along the guide shaft 7 in a direction orthogonal to the rotation axis C2 of the turntable 2, in the radial direction of the sample analysis disc 200, and in parallel with the sample analysis disc 200.

The optical pickup 20 includes an objective lens 21. The optical pickup 20 irradiates the laser beam 20a toward the sample analysis disc 200. The laser beam 20a is focused by the objective lens 21 on the track region 205 in which the reaction region 210 of the sample analysis disc 200 is formed.

With the sample analysis disc 200 rotated, the optical pickup 20 is driven in the radial direction of the sample analysis disc 200. As a result, as shown in FIG. 6, the laser beam 20a is scanned along the recess 204 corresponding to the track. The optical pickup 20 receives the reflected light from the sample analysis disc 200. The optical pickup 20 detects the light receiving level of the reflected light, generates a light receiving level signal JS, and outputs the light receiving level signal JS to the control unit 9.

The optical pickup drive circuit 8 controls the drive of the optical pickup 20. The optical pickup drive circuit 8 moves the optical pickup 20 along the guide shaft 7 and moves the objective lens 21 of the optical pickup 20 in the vertical direction.

The control unit 9 controls the turntable drive circuit 5 and the optical pickup drive circuit 8. The control unit 9 controls the turntable drive circuit 5 to rotate or stop the turntable 2 at a constant linear speed, for example. The control unit 9 controls the optical pickup drive circuit 8 to move the optical pickup 20 to a target position in the radial direction of the sample analysis disk 200, or to focus the laser beam 20a on the track region 205. Adjust the vertical position of the lens 21. As the control unit 9, for example, a CPU (Central Processing Unit) may be used.

The control unit 9 detects the reference position for each rotation cycle and track of the sample analysis disc 200 based on the reference position detection signal KS output from the reference position detection sensor 6. The control unit 9 identifies the reaction region 210 based on the detected reference position.

The storage unit 10 stores the measurement parameter SP1 for each track in each reaction region 210. The measurement parameter SP1 includes measurement information such as the number of reaction regions 210, the time corresponding to the distance from the notch 202 which is the reference position identification unit to each reaction region 210, and the timing of the measurement gate signal in each track. There is.

The control unit 9 reads the measurement parameter SP1 from the storage unit 10, and continuously generates a plurality of measurement gate signals GS1 for each track for each reaction region 210 based on the measurement parameter SP1. The control unit 9 extracts the fine particle pulse signal BS for each measurement gate signal GS1 from the light receiving level signal JS output from the optical pickup 20. The method of generating the measurement gate signal GS1 and the method of extracting the fine particle pulse signal BS will be described later.

The control unit 9 measures the number of fine particles 131 that label the substance to be detected 121 from the extracted fine particle pulse signal BS. The control unit 9 stores the number of fine particles 131 in each reaction region 210 in the storage unit 10 for each measurement gate signal GS1. The control unit 9 adds up the number of fine particles 131 for each reaction region 210 and displays them on the display unit 11. The number of fine particles 131 displayed corresponds to the number of substances 121 to be detected. A correction method for specifying the bubble region included in the reaction region 210 and correcting the measurement result of the fine particles 131 in the bubble region will be described later.

9 to 13, a method of analyzing the detection target substance 121 by the analyzer 1, specifically, a method of analyzing the fine particles 131 that label the detection target substance 121 will be described.

FIG. 9 schematically shows the positional relationship between the detection positions of the reference position detection sensor 6 and the optical pickup 20 and the notch 202 and the reaction region 210 of the sample analysis disc 200. The arrows in FIG. 9 indicate the rotation direction of the sample analysis disc 200. Reference numeral 6b indicates a detection position of the reference position detection sensor 6. The axis JL corresponds to the guide shaft 7.

In FIG. 9, the detection position 6b of the reference position detection sensor 6 is located on the axis JL, but the present invention is not limited to this. The detection position 6b can be any position on the outer peripheral portion of the sample analysis disc 200 as long as the cutout portion 202 can be detected. The optical pickup 20 moves along the axis JL in the radial direction of the sample analysis disc 200. Reference numeral 20b in FIG. 9 indicates a detection position of the optical pickup 20.

In the flowchart shown in FIG. 10, the control unit 9 controls the turntable drive circuit 5 so that the sample analysis disk 200 rotates at a constant linear velocity in step S1, and the turntable 2 is connected to the turntable drive unit 4. Is driven to rotate.

In step S2, the control unit 9 irradiates the detection light 6a from the reference position detection sensor 6 toward the sample analysis disc 200. In step S3, the control unit 9 irradiates the laser beam 20a from the optical pickup 20 toward the sample analysis disc 200. In addition, step S3 may be executed after step S2, step S2 may be executed after step S3, and step S2 and step S3 may be executed at the same time.

As shown in FIG. 9, the plurality of reaction regions 210 are formed so that their centers are evenly spaced on the same circumference with respect to the center C200 of the sample analysis disc 200. In order to distinguish the plurality of reaction regions 210, after the reference position detection sensor 6 detects the notch portion 202 with the detection light 6a, the code of the reaction region to which the laser light 20a is first irradiated is set to 211, and the laser light 20a is set. The symbols of the reaction regions to be sequentially irradiated with are 212, 213, 214, 215, 216, 217, 218.

FIG. 11 shows a reaction region 211 to which the laser beam 20a is first irradiated after the reference position detection sensor 6 detects the notch portion 202. The reaction regions 211 to 218 are irradiated with laser light 20a for each track from the tracks TRs located on the inner peripheral side of the sample analysis disk 200 toward the tracks TRe located on the outer peripheral side.

FIG. 11 shows a state in which the bubble region 221 is formed in the reaction region 211 over the track TRi-1, the track TRi, and the track TRi + 1. The bubble region 221 is formed by adhering the bubble 113, the bubble 123, or the bubble 133 on the track region 205 of the sample analysis disk 200 in step S101, step S102, or step S103 shown in FIG. In FIG. 11, the tracks TRs, TRi-1, TRi, TRi + 1, TRj-1, TRj, TRj + 1, and TRe are shown by straight lines for the sake of clarity.

The case where the laser beam 20a is scanned on the track TRi in the reaction region 211 including the bubble region 221 will be described below.

In step S4, the control unit 9 controls the optical pickup drive circuit 8 and moves the optical pickup 20 so that the track TRi of the sample analysis disc 200 is irradiated with the laser beam 20a. The reference position detection sensor 6 generates a reference position detection signal KS by detecting the notch portion 202 in step S5, and outputs the reference position detection signal KS to the control unit 9.

The optical pickup 20 receives the reflected light from the sample analysis disc 200 in step S6. The optical pickup 20 detects the light receiving level of the reflected light, generates a light receiving level signal JS, and outputs the light receiving level signal JS to the control unit 9.

In order to distinguish the reference position detection signal KS in each track TR, for example, the reference position detection signal in the track TRi is KSi, and the reference position detection signal in the track TRj is KSj. Further, in order to distinguish the measurement parameter SP1 in each track TR of each reaction region 210, for example, in the reaction region 211, the measurement parameter in the track TRi is set to SP11_i, and the measurement parameter in the track TRj is set to SP11_j.

In step S7, the control unit 9 detects the reference position detection signal KSi and reads out the measurement parameter SP11_i of the track TRi in the reaction region 211 from the storage unit 10.

In step S8, the control unit 9 measures the measurement gate signal GS11_i_1, GS11_i_2, GS11_i_3, GS11_i_4, GS11_i_5, which is a pulse signal for measuring the fine particles 131 of the track TRi in the reaction region 211 for each section based on the measurement parameter SP11_i. , GS11_i_6, GS11_i_7, GS11_i_8, GS11_i_9, GS11_i_10 are generated.

The measurement parameter SP11_i is the number of measurement gate signals GS1 in the track TRi of the reaction region 211, the time from the fall of the reference position detection signal KSi to the rise of the first measurement gate signal GS11_i_1, and the measurement gate signals GS11_i_1 to GS11_i_10. It contains measurement information such as each pulse width. The control unit 9 generates the measurement gate signal GS11_i_1 that rises when the time TD11_i elapses from the fall of the reference position detection signal KSi, and further continuously generates the measurement gate signals GS11_i_2 to GS11_i_10.

The measurement gate signals GS11_i_1 to GS11_i_10 are n concentric circles 2111,212, 211,2114 centered on the center C211 of the reaction region 211, and the center C200 and the reaction region of the sample analysis disk 200. It is generated at the timing of dividing the reaction region 211 in the track TRi into {(2 × n) + 2} by the dividing line SL211 passing through the center C211 of the 211. For example, as shown in FIG. 11, assuming that the diameters of the concentric circles 2111, 2112, 2113, 2114 are R1, R2, R3, R4 and the diameter of the reaction region 210 is R5, the relationship of R1 <R2 <R3 <R4 <R5. Has.

In step S101, step S102, or step S103 shown in FIG. 4, as described above, the bubble 113, the bubble 123, or the bubble 133 is a boundary portion between the inner surface and the bottom surface of the well 101, that is, a through hole of the cartridge 300. It easily adheres to the boundary between the inner peripheral surface of 301 and the track area 205 of the sample analysis disk 200. Therefore, the bubble region is likely to be formed on the outer peripheral portion of the reaction region 210. Therefore, it is preferable to set the measurement parameter SP1 so that the concentric circles 2111, 2112, 211 and 2114 are located on the outer peripheral side of the reaction region 210.

When the measurement parameter SP11_i is n = 4, 10 measurement gate signals GS1 are generated in the track TRi to TRj, and 8 measurement gate signals GS1 are generated in the track TRi-1 and the track TRj + 1. The TRs and the track TRe are set so that two measurement gate signals GS1 are generated.

In step S9, the control unit 9 outputs the fine particle pulse signal BS from the light receiving level signal JS output from the optical pickup 20 during the period (corresponding to the pulse width) from the rising point to the falling point of the measurement gate signal GS11_i_1. It is detected, the number of fine particles 131 is measured, and the storage unit 10 stores the number of the fine particles 131. The control unit 9 detects the fine particle pulse signal BS from the light receiving level signal JS output from the optical pickup 20 during each period from the rising point to the falling point of the measurement gate signal GS11_i_2 to GS11_i_10, and measures the number of fine particles 131. Then, the measurement result of the fine particles 131 is stored in the storage unit 10.

The light receiving level signal JS may contain noise in addition to the fine particle pulse signal BS. Therefore, the control unit 9 compares the pulse signal included in the light receiving level signal JS with the threshold value Vp, and determines that the pulse signal equal to or less than the threshold value Vp is the fine particle pulse signal BS.

The control unit 9 measures the number of fine particles 131 for each measurement gate signal GS1 for each reaction region 210 for all the track TRs from the tracks TRs to the tracks TRe in which the reaction regions 211 to 218 are formed, and the fine particles The measurement result of 131 is stored in the storage unit 10.

In step S10, the control unit 9 controls the turntable drive circuit 5 to stop the rotation of the sample analysis disc 200. The control unit 9 controls the reference position detection sensor 6 and the optical pickup 20 to stop the irradiation of the detection light 6a and the laser light 20a.

Next, with reference to FIGS. 11 to 13, a method of specifying the bubble region 221 by the analyzer 1 and a method of correcting the measurement result of the fine particles 131 in the bubble region 221 will be described.

As shown in FIG. 11, a method for identifying the bubble region 221 by the analyzer 1 when the bubble region 221 is formed over the track TRi-1, the track TRi, and the track TRi + 1 in the reaction region 211, and the bubble region. A method of correcting the measurement result of the fine particles 131 in 221 will be described.

In the bubble region 221, the fine particle pulse signal BS is not detected because the fine particles 131 are not captured on the track region 205. For example, in the track TRi, the fine particle pulse signal BS is not detected in the period from the rising point to the falling point of the measurement gate signal GS11_i_1 corresponding to the measurement position of the bubble region 221.

In the flowchart shown in FIG. 12, in step S11, the control unit 9 reads out the measurement result for each measurement gate signal GS1 from the storage unit 10 for each reaction region 210. The control unit 9 reads, for example, the measurement result of the fine particles 131 for each measurement gate signal GS1 in the reaction region 211 from the storage unit 10.

In step S12, the control unit 9 compares the measurement results having a symmetrical positional relationship in the reaction region 210. Hereinafter, in the reaction region 211, the case where the measurement result by the measurement gate signal GS11_i_1 in the track TRi and the measurement result by the measurement gate signal GS1 at the measurement position symmetrical to the measurement position of the measurement gate signal GS11_i_1 are compared. explain.

The measurement gate signal GS11_i_1 (first measurement gate signal) and the measurement gate signal GS11_i_10 (second measurement gate signal) have a symmetrical positional relationship with respect to the dividing line SL211 and have the same pulse width. Further, the measurement gate signal GS11_i_1 and the measurement gate signal GS11_i_1 have the same measurement position from the center C211 of the reaction region 211. Therefore, the measurement result obtained by the measurement gate signal GS11_i_1 and the measurement result obtained by the measurement gate signal GS11_i_1 have a symmetrical positional relationship with respect to the dividing line SL211.

The track TRi and the track TRj are in a symmetrical positional relationship with respect to the axis of symmetry AS211 which passes through the center C211 of the reaction region 211 and is orthogonal to the dividing line SL211. The control unit 9 is a pulse signal for measuring the fine particles 131 of the track TRj in the reaction region 211 for each section based on the measurement parameter SP11_j. Generates GS11_j_8, GS11_j_9, and GS11_j_10. The control unit 9 generates the measurement gate signal GS11_j_1 that rises when the time TD11_j elapses from the fall of the reference position detection signal KSj, and further continuously generates the measurement gate signals GS11_j_2 to GS11_j_10.

The measurement gate signal GS11_i_1 and the measurement gate signal GS11_j_1 (third measurement gate signal) have a symmetrical positional relationship with respect to the axis of symmetry AS211 and have the same pulse width. Further, the measurement gate signal GS11_i_1 and the measurement gate signal GS11_j_1 have the same measurement position from the center C211 of the reaction region 211. Therefore, the measurement result obtained by the measurement gate signal GS11_i_1 and the measurement result obtained by the measurement gate signal GS11_j_1 have a symmetrical positional relationship with respect to the axis of symmetry AS211.

Similarly, the measurement gate signal GS11_i_10 and the measurement gate signal GS11_j_10 (fourth measurement gate signal) have a symmetrical positional relationship with respect to the axis of symmetry AS211 and have the same pulse width. Further, the measurement gate signal GS11_i_10 and the measurement gate signal GS11_j_10 have the same measurement position from the center C211 of the reaction region 211. Therefore, the measurement result obtained by the measurement gate signal GS11_i_10 and the measurement result obtained by the measurement gate signal GS11_j_10 have a symmetrical positional relationship with respect to the axis of symmetry AS211.

Therefore, the measurement results obtained by the measurement gate signal GS11_i_1, the measurement gate signal GS11_i_10, the measurement gate signal GS11_j_1, and the measurement gate signal GS11_j_10 have a symmetrical positional relationship. The control unit 9 compares the measurement results obtained by each measurement gate signal GS11_i_1, GS11_i_10, GS11_j_1, GS11_j_10.

Specifically, the control unit 9 calculates the average value of the number of fine particles 131 obtained by each measurement gate signal GS11_i_1, GS11_i_10, GS11_j_1, GS11_j_10. When the bubble region 221 is not provided, the number of fine particles 131 obtained by each measurement gate signal GS11_i_1, GS11_i_10, GS11_j_1, GS11_j_10 is usually about the same value.

The control unit 9 determines that the region corresponding to the measurement gate signal GS1 in which the number of fine particles 131 having a predetermined ratio or less with respect to the average value is obtained is the bubble region. Since the fine particles 131 are not captured at the measurement position corresponding to the measurement gate signal GS11_i_1, the control unit 9 determines that the region corresponding to the measurement gate signal GS11_i_1 is the bubble region.

Since the fine particles 131 are not captured by the bubbles 113, 123, 133 in the bubble region 221, it is desirable to correct the measurement result. That is, the bubble region 221 is a measurement result correction target region for correcting the number of fine particles 131. The number of fine particles 131 obtained by the measurement gate signal GS1 is affected by the number of detection target substances 121 contained in the sample liquid 122. Therefore, it is desirable to determine the bubble region by the ratio rather than the absolute value of the number of fine particles 131.

In step S13, the control unit 9 compares the measurement results of adjacent tracks TR in the reaction region 210. Normally, in step S101, step S102, or step S103, the bubbles 113, 123, and 133 adhering to the track region 205 of the sample analysis disk 200 are formed over the plurality of track TRs. As shown in FIG. 13, the measurement positions of the measurement gate signal GS11_i-1_1 on the track TRi-1, the measurement gate signal GS11_i_1 on the track TRi, and the measurement gate signal GS11_i + 1_1 on the track TRi + 1 are continuous along the outer periphery of the reaction region 211. Has a positional relationship. FIG. 13 corresponds to FIG.

The control unit 9 compares the measurement results obtained by the measurement gate signals GS11_i-1_1, GS11_i_1, and GS11_i + 1_1 in the adjacent tracks TRi-1, track TRi, and track TRi + 1 in the reaction region 211, for example.

If the region corresponding to each measurement gate signal GS11_i-1_1, GS11_i_1, GS11_i + 1_1 is determined to be a bubble region in step S12, the control unit 9 controls the region corresponding to each measurement gate signal GS11_i-1_1, GS11_i_1, GS11_i + 1_1. Is identified as the bubble region 221 formed over track TRi-1, track TRi, and track TRi + 1.

In step S14, the control unit 9 corrects the measurement result of the fine particles 131 in the bubble region 221. The control unit 9 calculates, for example, the average value of the number of fine particles 131 obtained by each measurement gate signal GS11_i_10, GS11_j_1, GS11_j_10 that was not determined to be the bubble region 221 and is determined to be the bubble region 221. The number of fine particles 131 obtained by the gate signal GS11_i_1 is corrected to the calculated average value.

Similarly, the control unit 9 calculates the average value of the number of fine particles 131 obtained by each measurement gate signal GS11_i-1_10, GS11_j + 1_11, GS11_j + 1_10, and determines the number of fine particles 131 obtained by the measurement gate signal GS11_i-1_1. Correct to the calculated average value. Further, the control unit 9 calculates the average value of the number of fine particles 131 obtained by each measurement gate signal GS11_i + 1_10, GS11_j-1_11, GS11_j-1_10, and calculates the number of fine particles 131 obtained by the measurement gate signal GS11_i + 1_1. Correct to the average value.

That is, the area 232 corresponding to the measurement gate signal GS11_i-1_10, GS11_i_10, GS11_i + 1_10, the area 233 corresponding to the measurement gate signal GS11_j-1_1, GS11_j_1, GS11_j + 1_1, the measurement gate signal GS11_j-1_10, GS11_j_10, GS11_10 This is a comparison target region for correcting the number of fine particles 131 in the bubble region 221.

The control unit 9 executes steps S11 to S14 for all the tracks TRs to TRes in all the reaction regions 210 (211-218).

In step S15, the control unit 9 causes the display unit 11 to display the measurement result and the correction result of the fine particles 131 in each reaction region 210.

According to the analyzer 1 and the analysis method of the first embodiment, the bubble region 221 is specified by comparing the measurement results having a symmetrical positional relationship in the reaction region 210 and further comparing the measurement results of adjacent tracks. be able to. Further, among the measurement results having a symmetrical positional relationship, the measurement result determined to be the bubble region 221 can be corrected based on the measurement result not determined to be the bubble region.

[Second Embodiment]
The analyzer and analysis method of the second embodiment will be described with reference to FIGS. 8, 10, 12, 14, and 15. As shown in FIG. 8, the analyzer 1001 of the second embodiment is provided with the control unit 1009 and the storage unit 1010 instead of the control unit 9 and the storage unit 10, and the method for analyzing fine particles in the analyzer 1001 is an analyzer. It is different from the method for analyzing fine particles in 1. Therefore, a method for analyzing fine particles in the analyzer 1001 will be described. In addition, in order to make the explanation easy to understand, the same components as those of the analyzer 1 of the first embodiment are designated by the same reference numerals.

When the substance 121 to be detected is an exosome, the size of the exosome is as small as about 100 nm, so that it is difficult to directly detect it optically. The analyzer 1001 of the second embodiment indirectly detects the detection target substance 121 specifically bound to the fine particles 131 by detecting and measuring the fine particles 131 captured in the reaction region 210. measure.

The control unit 1009 controls the turntable drive circuit 5 and the optical pickup drive circuit 8. The control unit 1009 controls the turntable drive circuit 5 to rotate or stop the turntable 2 at a constant linear speed, for example. The control unit 1009 controls the optical pickup drive circuit 8 to move the optical pickup 20 to a target position in the radial direction of the sample analysis disk 200, or an objective so that the laser beam 20a is focused on the track region 205. Adjust the vertical position of the lens 21. For example, a CPU may be used as the control unit 1009.

The control unit 1009 detects the reference position for each rotation cycle and track of the sample analysis disc 200 based on the reference position detection signal KS output from the reference position detection sensor 6. The control unit 1009 identifies the reaction region 210 based on the detected reference position.

The storage unit 1010 stores the measurement parameter SP2 for each track in each reaction region 210. The measurement parameter SP2 includes measurement information such as the number of reaction regions 210, the time corresponding to the distance from the notch 202 which is the reference position identification unit to each reaction region 210, and the timing of the measurement gate signal in each track. There is.

The control unit 1009 reads out the measurement parameter SP2 from the storage unit 1010, and continuously generates a plurality of measurement gate signals GS2 for each track for each reaction region 210 based on the measurement parameter SP2. The control unit 1009 extracts a fine particle pulse signal BS for each measurement gate signal GS2 from the light receiving level signal JS output from the optical pickup 20. The method of generating the measurement gate signal GS2 and the method of extracting the fine particle pulse signal BS will be described later.

The control unit 1009 measures the number of fine particles 131 that label the substance to be detected 121 from the extracted fine particle pulse signal BS. The control unit 1009 stores the number of fine particles 131 in each reaction region 210 in the storage unit 1010 for each measurement gate signal GS2. The control unit 1009 adds up the number of fine particles 131 for each reaction region 210 and displays them on the display unit 11. The number of fine particles 131 displayed corresponds to the number of substances 121 to be detected. A correction method for specifying the bubble region included in the reaction region 210 and correcting the measurement result of the fine particles 131 in the bubble region will be described later.

The control unit 1009 measures the number of fine particles 131 that label the substance to be detected 121 from the extracted fine particle pulse signal BS. The control unit 1009 stores the number of fine particles 131 in each reaction region 210 in the storage unit 1010 for each measurement gate signal GS2. The control unit 1009 adds up the number of fine particles 131 for each reaction region 210 and displays them on the display unit 11. The number of fine particles 131 displayed corresponds to the number of substances 121 to be detected. A correction method for specifying the bubble region included in the reaction region 210 and correcting the measurement result of the fine particles 131 in the bubble region will be described later.

A method of analyzing the detection target substance 121 by the analyzer 1001 and specifically, a method of analyzing the fine particles 131 that label the detection target substance 121 will be described with reference to FIGS. 10, 12, 14, and 15.

In the flowchart shown in FIG. 10, the control unit 1009 controls the turntable drive circuit 5 so that the sample analysis disk 200 rotates at a constant linear velocity in step S21, and the turntable 2 is connected to the turntable drive unit 4. Is driven to rotate.

In step S22, the control unit 1009 irradiates the detection light 6a from the reference position detection sensor 6 toward the sample analysis disk 200. In step S23, the control unit 1009 irradiates the laser beam 20a from the optical pickup 20 toward the sample analysis disc 200. In addition, step S23 may be executed after step S22, step S22 may be executed after step S23, or step S22 and step S23 may be executed at the same time.

FIG. 14 shows a reaction region 211 to which the laser beam 20a is first irradiated after the reference position detection sensor 6 detects the notch portion 202. The reaction regions 211 to 218 are irradiated with laser light 20a for each track from the tracks TRs located on the inner peripheral side of the sample analysis disk 200 toward the tracks TRe located on the outer peripheral side.

FIG. 14 shows a state in which the bubble region 241 is formed in the reaction region 211 over the track TRi-1, the track TRi, and the track TRi + 1. The bubble region 241 is formed by adhering the bubble 113, the bubble 123, or the bubble 133 on the track region 205 of the sample analysis disk 200 in step S101, step S102, or step S103 shown in FIG. In FIG. 14, the tracks TRs, TRi-1, TRi, TRi + 1, TRj-1, TRj, TRj + 1, and TRe are shown by straight lines for the sake of clarity.

The case where the laser beam 20a is scanned on the track TRi in the reaction region 211 including the bubble region 241 will be described below.

In step S24, the control unit 1009 controls the optical pickup drive circuit 8 and moves the optical pickup 20 so that the track TRi of the sample analysis disk 200 is irradiated with the laser beam 20a. The reference position detection sensor 6 generates a reference position detection signal KS by detecting the notch portion 202 in step S25, and outputs the reference position detection signal KS to the control unit 1009.

The optical pickup 20 receives the reflected light from the sample analysis disc 200 in step S26. The optical pickup 20 detects the light receiving level of the reflected light, generates a light receiving level signal JS, and outputs the light receiving level signal JS to the control unit 1009.

In order to distinguish the measurement parameter SP2 in each track TR of each reaction region 210, for example, in the reaction region 211, the measurement parameter in the track TRi is SP21_i, and the measurement parameter in the track TRj is SP21_j.

In step S27, the control unit 1009 detects the reference position detection signal KSi and reads the measurement parameter SP21_i of the track TRi in the reaction region 211 from the storage unit 1010.

In step S28, the control unit 1009 measures gate signals GS21_i_1, GS21_i_2, GS21_i_3, GS21_i_4, GS21_i_5, which are pulse signals for measuring the fine particles 131 of the track TRi in the reaction region 211 for each section based on the measurement parameter SP21_i. , GS21_i_6, GS21_i_7, GS21_i_8, GS21_i_9, GS21_i_10 are generated.

The measurement parameter SP21_i is the number of measurement gate signals GS2 in the track TRi of the reaction region 211, the time TD21_i from the fall of the reference position detection signal KSi to the rise of the first measurement gate signal GS21_i_1, and the measurement gate signals GS21_i_1 to GS21_i_10. It contains measurement information such as each pulse width. The control unit 1009 generates the measurement gate signal GS21_i_1 that rises when the time TD21_i elapses from the fall of the reference position detection signal KSi, and further continuously generates the measurement gate signals GS21_i_2 to GS21_i_10.

The measurement gate signals GS21_i_1 to GS21_i_10 are the reaction regions in each track TR by n ellipses (2116, 2117, 2118, 2119) centered on the center C211 of the reaction region 211 and the dividing line SL211. It is generated at the timing of dividing 211 into {(2 × n) + 2} pieces.

The major axes of the ellipses 2116, 2117, 2118, and 2119 are arranged on the dividing line SL211. The major axis ends of the ellipses 2116, 2117, 2118, and 2119 are located on the outer circumference of the reaction region 210. For example, as shown in FIG. 14, if the minor axis of the ellipses 2116, 2117, 2118, 2119 is R11, R12, R13, R14, the major axis is R21, R22, R23, R24, and the diameter of the reaction region 210 is R5. , R11 <R12 <R13 <R14 <R5, and R21 = R22 = R23 = R24 = R5. That is, the ellipses 2116, 2117, 2118, and 2119 have the same major axis and different minor axis. Therefore, when n = 4, 10 measurement gate signals GS2 are generated for each of the tracks TR (TRs to TRe).

In step S101, step S102, or step S103 shown in FIG. 4, as described above, the bubble 113, the bubble 123, or the bubble 133 is a boundary portion between the inner surface and the bottom surface of the well 101, that is, a through hole of the cartridge 300. It easily adheres to the boundary between the inner peripheral surface of 301 and the track area 205 of the sample analysis disk 200. Therefore, the bubble region is likely to be formed on the outer peripheral portion of the reaction region 210. Therefore, it is preferable to set the measurement parameter SP2 so that the short axis ends of the ellipses 2116, 2117, 2118, and 2119 are located on the outer peripheral side of the reaction region 210.

In step S29, the control unit 1009 outputs the fine particle pulse signal BS from the light receiving level signal JS output from the optical pickup 20 during the period (corresponding to the pulse width) from the rising point to the falling point of the measurement gate signal GS21_i_1. Detected, the number of fine particles 131 is measured and stored in the storage unit 1010. The control unit 1009 detects the fine particle pulse signal BS from the light receiving level signal JS output from the optical pickup 20 during each period from the rising point to the falling point of the measurement gate signals GS21_i_2 to GS21_i_10, and measures the number of fine particles 131. Then, the measurement result of the fine particles 131 is stored in the storage unit 1010.

The light receiving level signal JS may contain noise in addition to the fine particle pulse signal BS. Therefore, the control unit 1009 compares the pulse signal included in the light receiving level signal JS with the threshold value Vp, and determines that the pulse signal equal to or less than the threshold value Vp is the fine particle pulse signal BS.

The control unit 1009 measures the number of fine particles 131 for each measurement gate signal GS2 for each reaction region 210 for all the track TRs from the tracks TRs to the tracks TRe in which the reaction regions 211 to 218 are formed, and the fine particles The measurement result of 131 is stored in the storage unit 1010.

In step S30, the control unit 1009 controls the turntable drive circuit 5 to stop the rotation of the sample analysis disk 200. The control unit 1009 controls the reference position detection sensor 6 and the optical pickup 20 to stop the irradiation of the detection light 6a and the laser light 20a.

Next, a method of specifying the bubble region 241 by the analyzer 1001 and a method of correcting the measurement result of the fine particles 131 in the bubble region 241 will be described with reference to FIGS. 12, 14, and 15.

As shown in FIG. 14, a method for identifying the bubble region 241 by the analyzer 1001 when the bubble region 241 is formed over the track TRi-1, the track TRi, and the track TRi + 1 in the reaction region 211, and the bubble region. A method of correcting the measurement result of the fine particles 131 in 241 will be described.

In the bubble region 241 the fine particle 131 is not captured on the track region 205, so that the fine particle pulse signal BS is not detected. For example, in the track TRi, the fine particle pulse signal BS is not detected in the period from the rising point to the falling point of the measurement gate signal GS21_i_1 corresponding to the measurement position of the bubble region 241.

In the flowchart shown in FIG. 12, in step S31, the control unit 1009 reads out the measurement result for each measurement gate signal GS2 from the storage unit 1010 for each reaction region 210. The control unit 1009 reads, for example, the measurement result of the fine particles 131 for each measurement gate signal GS2 in the reaction region 211 from the storage unit 1010.

In step S32, the control unit 1009 compares the measurement results having a symmetrical positional relationship in the reaction region 210. Below, in the reaction region 211, the case where the measurement result by the measurement gate signal GS21_i_1 in the track TRi and the measurement result by the measurement gate signal GS2 at the measurement position symmetrical to the measurement position of the measurement gate signal GS21_i_1 are compared. explain.

The measurement gate signal GS21_i_1 (fifth measurement gate signal) and the measurement gate signal GS21_i_10 (sixth measurement gate signal) have a symmetrical positional relationship with respect to the dividing line SL211 and have the same pulse width. Further, the measurement gate signal GS21_i_1 and the measurement gate signal GS21_i_1 have the same measurement position from the center C211 of the reaction region 211. Therefore, the measurement result obtained by the measurement gate signal GS21_i_1 and the measurement result obtained by the measurement gate signal GS21_i_1 have a symmetrical positional relationship with respect to the dividing line SL211.

The track TRi and the track TRj are in a symmetrical positional relationship with respect to the axis of symmetry AS211 which passes through the center C211 of the reaction region 211 and is orthogonal to the dividing line SL211. The control unit 1009 is a pulse signal for measuring the fine particles 131 of the track TRj in the reaction region 211 for each section based on the measurement parameter SP21_j. Generates GS21_j_8, GS21_j_9, and GS21_j_10. The control unit 1009 generates the measurement gate signal GS21_j_1 that rises when the time TD21_j elapses from the fall of the reference position detection signal KSj, and further continuously generates the measurement gate signals GS21_j_2 to GS21_j_10.

The measurement gate signal GS21_i_1 and the measurement gate signal GS21_j_1 (seventh measurement gate signal) have a symmetrical positional relationship with respect to the axis of symmetry AS211 and have the same pulse width. Further, the measurement gate signal GS21_i_1 and the measurement gate signal GS21_j_1 have the same measurement position from the center C211 of the reaction region 211. Therefore, the measurement result obtained by the measurement gate signal GS21_i_1 and the measurement result obtained by the measurement gate signal GS21_j_1 have a symmetrical positional relationship with respect to the axis of symmetry AS211.

Similarly, the measurement gate signal GS21_i_10 and the measurement gate signal GS21_j_10 (eighth measurement gate signal) have a symmetrical positional relationship with respect to the axis of symmetry AS211 and have the same pulse width. Further, the measurement gate signal GS21_i_10 and the measurement gate signal GS21_j_10 have the same measurement position from the center C211 of the reaction region 211. Therefore, the measurement result obtained by the measurement gate signal GS21_i_10 and the measurement result obtained by the measurement gate signal GS21_j_10 have a symmetrical positional relationship with respect to the axis of symmetry AS211.

Therefore, the measurement results obtained by the measurement gate signal GS21_i_1, the measurement gate signal GS21_i_10, the measurement gate signal GS21_j_1, and the measurement gate signal GS21_j_10 have a symmetrical positional relationship. The control unit 1009 compares the measurement results obtained by each measurement gate signal GS21_i_1, GS21_i_10, GS21_j_1, GS21_j_10.

Specifically, the control unit 1009 calculates the average value of the number of fine particles 131 obtained by each measurement gate signal GS21_i_1, GS21_i_10, GS21_j_1, GS21_j_10. When the bubble region 241 is not provided, the number of fine particles 131 obtained by each measurement gate signal GS21_i_1, GS21_i_10, GS21_j_1, GS21_j_10 is usually about the same value.

The control unit 1009 determines that the region corresponding to the measurement gate signal GS2 in which the number of fine particles 131 having a predetermined ratio or less with respect to the average value is obtained is the bubble region. Since the fine particles 131 are not captured at the measurement position corresponding to the measurement gate signal GS21_i_1, the control unit 1009 determines that the region corresponding to the measurement gate signal GS21_i_1 is the bubble region.

Since the fine particles 131 are not captured by the bubbles 113, 123, 133 in the bubble region 241, it is desirable to correct the measurement result. That is, the bubble region 241 is a measurement result correction target region for correcting the number of fine particles 131. The number of fine particles 131 obtained by the measurement gate signal GS2 is affected by the number of detection target substances 121 contained in the sample liquid 122. Therefore, it is desirable to determine the bubble region by the ratio rather than the absolute value of the number of fine particles 131.

In step S33, the control unit 1009 compares the measurement results of adjacent tracks TR in the reaction region 210. Normally, in step S101, step S102, or step S103, the bubbles 113, 123, and 133 adhering to the track region 205 of the sample analysis disk 200 are formed over the plurality of track TRs. As shown in FIG. 15, the measurement positions of the measurement gate signal GS21_i-1_1 on the track TRi-1, the measurement gate signal GS21_i_1 on the track TRi, and the measurement gate signal GS21_i + 1_1 on the track TRi + 1 are continuous along the outer periphery of the reaction region 211. Has a positional relationship. FIG. 15 corresponds to FIG.

The control unit 1009 compares the measurement results obtained by the measurement gate signals GS21_i-1_1, GS21_i_1, and GS21_i + 1_1 in the adjacent tracks TRi-1, track TRi, and track TRi + 1 in the reaction region 211, for example.

In step S32, when the region corresponding to each measurement gate signal GS21_i-1_1, GS21_i_1, GS21_i + 1_1 is determined to be a bubble region, the control unit 1009 determines the region corresponding to each measurement gate signal GS21_i-1_1, GS21_i_1, GS21_i + 1_1. Is identified as the bubble region 241 formed over track TRi-1, track TRi, and track TRi + 1.

In step S34, the control unit 1009 corrects the measurement result of the fine particles 131 in the bubble region 241. The control unit 1009 calculates, for example, the average value of the number of fine particles 131 obtained by each measurement gate signal GS21_i_10, GS21_j_1, GS21_j_10 that is not determined to be the bubble region 241 and measures determined to be the bubble region 241. The number of fine particles 131 obtained by the gate signal GS21_i_1 is corrected to the calculated average value.

Similarly, the control unit 1009 calculates the average value of the number of fine particles 131 obtained by each measurement gate signal GS21_i-1_10, GS21_j + 1_1, GS21_j + 1_10, and determines the number of fine particles 131 obtained by the measurement gate signal GS21_i-1_1. Correct to the calculated average value. Further, the control unit 1009 calculates the average value of the number of fine particles 131 obtained by each measurement gate signal GS21_i + 1_10, GS21_j-1_11, GS21_j-1_10, and calculates the number of fine particles 131 obtained by the measurement gate signal GS21_i + 1_1. Correct to the average value.

That is, the area 252 corresponding to the measurement gate signal GS21_i-1_10, GS21_i_10, GS21_i + 1_10, the area 253 corresponding to the measurement gate signal GS21_j-1_1, GS21_j_1, GS21_j + 1_1, the measurement gate signal GS21_j-1_10, GS21_j_10, GS21_10 This is a comparison target region for correcting the number of fine particles 131 in the bubble region 241.

The control unit 1009 executes steps S31 to S34 for all tracks TRs to TRes in all reaction regions 210 (211-218).

In step S35, the control unit 1009 causes the display unit 11 to display the measurement result and the correction result of the fine particles 131 in each reaction region 210.

According to the analyzer 1001 and the analysis method of the second embodiment, the bubble region 241 is specified by comparing the measurement results having a symmetrical positional relationship in the reaction region 210 and further comparing the measurement results of adjacent tracks. be able to. Further, among the measurement results having a symmetrical positional relationship, the measurement result determined to be the bubble region 241 can be corrected based on the measurement result not determined to be the bubble region.

In the analyzer 1 of the first embodiment, the number of measurement gate signals GS differs depending on the track TR, whereas in the analyzer 1001 of the second embodiment, the number of measurement gate signals GS is the same in all track TRs. Since there is a possibility that the measurement accuracy of the fine particles 131 and the correction accuracy in the bubble region may decrease in the track TR having a small number of measurement gate signal GS, it is preferable that the number of measurement gate signal GS is the same in all track TRs.

The present invention is not limited to the above-described embodiment, and various modifications can be made without departing from the gist of the present invention.

For example, in steps S13 and S33, the measurement results are not compared for all adjacent tracks, but when it is determined in steps S12 and S32 that the measurement results are in the bubble region, the track is determined to be in the bubble region. The measurement results may be compared for adjacent tracks.

The number of concentric circles for generating the measurement gate signal GS1 and the diameter of each concentric circle are not limited to the above-described first embodiment, and may be set to any value.

For example, the measurement parameter SP1 is set so that the intervals between the concentric circles 2111 to 2114 with respect to the reaction region 210 are equal to (R2-R1) = (R3-R2) = (R4-R3) = (R5-R4). May be good. Further, in order to improve the determination accuracy (resolution) of the bubble region formed on the outer peripheral portion of the reaction region 210, the intervals between the concentric circles 2111 to 2114 are (R2-R1)> (R3-R2)> (R4-R3)>. The measurement parameter SP1 may be set so as to narrow toward (R5-R4) and the outer circumference.

The number of ellipses for generating the measurement gate signal GS2 and the minor axis of each ellipse are not limited to the second embodiment described above, and may be set to any value.

1,1001 Analyzer 2 Turntable 4 Turntable drive 5 Turntable drive 8 Optical pickup drive circuit 9,1009 Control 20 Optical pickup 20a Laser light 121 Detectable substance 131 Fine particles 200 Sample analysis disk 210 Reaction area 221, 241 Bubble area (area to be corrected for measurement results)
C2 rotation axis JS light receiving level signal GS1, GS2 measurement gate signal

Claims (11)

A turntable that holds a sample analysis disc with a reaction region in which fine particles that bind to the substance to be detected are captured for each track.
The turntable drive unit that rotates the turntable and
A turntable drive circuit that controls the turntable drive unit,
An optical pickup that is driven along a direction orthogonal to the rotation axis of the turntable, irradiates the reaction region with laser light, receives reflected light from the reaction region, and generates a light receiving level signal.
An optical pickup drive circuit that controls the drive of the optical pickup,
A control unit that controls the turntable drive circuit and the optical pickup drive circuit is provided.
The control unit
A plurality of measurement gate signals for measuring the number of the fine particles captured in the reaction region are continuously generated for each track, and the number of the fine particles is measured for each measurement gate signal from the light receiving level signal. compares the number of each of the particles measured before each Symbol measurement gate signal which is in a symmetrical position relationship with respect to the dividing line passing through the center of the reaction region, the measurement result for correcting the number of the fine particles Identify the area to be corrected
Analysis apparatus. The control unit divides the reaction region in each track by a plurality of concentric circles centered on the center of the reaction region and a dividing line passing through the center of the sample analysis disk and the center of the reaction region. analyzer according to請Motomeko 1 that generates a plurality of measurement gate signal. The control unit corresponds to the first measurement gate signal in the plurality of measurement gate signals and the measurement position having a symmetrical positional relationship with the measurement position corresponding to the first measurement gate signal with respect to the dividing line. Corresponds to the second measurement gate signal and the measurement position having a symmetrical positional relationship with the measurement position corresponding to the first measurement gate signal with respect to the axis of symmetry passing through the center of the reaction region and orthogonal to the dividing line. Each measurement gate signal of the third measurement gate signal and the fourth measurement gate signal corresponding to the measurement position having a symmetrical positional relationship with the measurement position corresponding to the second measurement gate signal with respect to the axis of symmetry. analyzer according to請Motomeko 2 that identifies the measurement result correction target region by comparing the number of each of the particles measured in the period. Wherein the control unit, analyzer according to請Motomeko 3 that identifies the measurement result correction target region by comparing the number of each of the fine particles measured during the measurement gate signal of adjacent tracks. When the measurement position corresponding to the first measurement gate signal is specified as the measurement result correction target region, the control unit determines the second measurement gate signal, the third measurement gate signal, and the measurement gate signal. fourth calculating the average number of each of the particles measured in the period of each measurement gate signal of the measuring gate signal, the number of the particles measured in the period of the first measurement gate signal spectrometer according to請Motomeko 3 or 4 correct the average value. The control unit has a center point of the reaction region, and each track is formed by a plurality of ellipses having the same major axis and a different minor axis, and a dividing line passing through the center of the sample analysis disk and the center of the reaction region. analyzer according to請Motomeko 1 that generates a plurality of measurement gate signal at a timing that divides the reaction zone in. Wherein the plurality of major axis of the ellipse is located on the dividing line, the major axis end of said plurality of ellipse analyzer according to請Motomeko 6 that is located on the outer periphery of the reaction region. The control unit corresponds to the fifth measurement gate signal in the plurality of measurement gate signals and the measurement position having a symmetrical positional relationship with the measurement position corresponding to the fifth measurement gate signal with respect to the dividing line. Corresponds to the sixth measurement gate signal and the measurement position having a symmetrical positional relationship with the measurement position corresponding to the fifth measurement gate signal with respect to the axis of symmetry passing through the center of the reaction region and orthogonal to the dividing line. Each measurement gate signal of the seventh measurement gate signal and the eighth measurement gate signal corresponding to the measurement position having a symmetrical positional relationship with the measurement position corresponding to the sixth measurement gate signal with respect to the axis of symmetry. analyzer according to請Motomeko 6 or 7 that identifies the measurement result correction target region by comparing the number of each of the particles measured in the period. The analyzer according to claim 8, wherein the control unit specifies the measurement result correction target region by comparing the number of the fine particles measured during the period of the measurement gate signals of adjacent tracks. .. When the measurement position corresponding to the fifth measurement gate signal is specified as the measurement result correction target region, the control unit determines the sixth measurement gate signal, the seventh measurement gate signal, and the measurement gate signal. calculates an average value of the number of each of the particles measured in the period of each measurement gate signal of the eighth measurement gate signal, the number of the fifth measurement gate signal the particles measured in a period of analyzer according to請Motomeko 8 or 9 you correction to the average value. Rotate the sample analysis disc, which has a reaction region in which fine particles bound to the substance to be detected are captured for each track.
The reaction region is irradiated with laser light for each track.
The reflected light from the reaction region is received to generate a light receiving level signal.
A plurality of measurement gate signals for measuring the number of the fine particles captured in the reaction region are continuously generated for each track.
From the light receiving level signal, the number of the fine particles is measured for each measurement gate signal, and the number of the fine particles is measured.
Measurement result correction target for comparing the number of each of the fine particles measured for each measurement gate signal having a symmetrical positional relationship with respect to the dividing line passing through the center of the reaction region and correcting the number of the fine particles. Identify the area
Minute析方method.

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