JP2012237711A - Optical analysis device, and optical analysis method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To enable the detection of the concentration of antigens and antibodies contained in a sample to be analyzed with high quantitativity in a wide range from low concentration to high concentration.SOLUTION: An optical analysis device 10 includes: a first track configured by a groove 107 or a land 108 provided in a disk 100 for sample analysis; a pickup 12 for irradiating second and third tracks adjacent across the first track with a laser beam to acquire reflected light; a reflected light detecting part 128 for simultaneously detecting reflected light of the first to third tracks acquired by the pickup 12; and a sample detecting part 18 for detecting the amount of biopolymers connected to a bead 110 for a marker fixed on the disk 100 for sample analysis by the fluctuation of the reflected light in the first track and the fluctuation of the reflected light in one between the second and third tracks detected by the reflected light detecting part 128.

Description

  The present invention relates to an optical analyzer and an optical analysis method for analyzing a biopolymer such as an antigen or an antibody contained in a biological sample such as blood by an optical method.

  In recent years, immunoassays for detecting antigens and antibodies from biological samples contained in blood or the like have been used in various situations for the purpose of early diagnosis of diseases in disease diagnosis and health checkup. An immunoassay is a method in which a measurable label is attached to an antibody (or antigen) that specifically binds to a specific antigen (or antibody) contained in a biological sample, and the result of the reaction of both is quantified. In this method, the concentration of the antigen (or antibody) contained in the sample is determined to determine the disease.

  Since this method is relatively sensitive and easy to operate, various labeling methods have been developed. For example, there are RIA method (radioinom assay) using radioisotope for labeling, EIA method (enzyme immunoassay) using enzyme, FIA method using fluorescent label, CLIA method using chemiluminescence.

  Among them, an ELISA method (Enzyme-Linked ImmunoSorbent Assay), which is one of the methods using an enzyme, is widely and widely used as a test method using a microplate. The method using a microplate can measure a large number of samples at a time using a 96-well plate and a general-purpose microplate reader, and is relatively inexpensive. In addition, compared to the RIA method that uses radioactive materials, there are advantages such as fewer restrictions on the place to use. However, pretreatment to fix the antibody on the plate, reaction time to react the antibody and antigen, B / F (bond / free) separation to wash away the unreacted label, enzyme reaction of the label, etc. And the total inspection time is from several hours to one day.

  For this reason, in order to shorten the inspection time, an analysis chip (Lab on a chip) has been developed by using microfabrication technology and replacing the operation procedure in each well of the microplate on a chip of several centimeters square. Yes. The processing time is shortened by preparing antibodies on the chip and immobilizing antibodies and preparing pre-treatments for general ELISA in advance. In addition, the reaction time is shortened by performing the antigen-antibody reaction in a narrow channel of several tens of micrometers to several millimeters. This makes it possible to shorten the inspection time by several tens of minutes by using a dedicated analysis chip for each inspection. Conventionally, after taking a patient sample, it took a long time to send it to the laboratory, but in some examinations such as myocardial markers, examinations in the examination room or the patient's bedside, so-called Devices capable of POCT (Point of Care Test) are being developed.

Special table 2004-533606 gazette JP-T-2006-521558 JP-T-2002-530786

  Patent Document 1 and Patent Document 2 disclose a disc-shaped analysis device having a feature that combines the advantages of a microplate capable of simultaneously testing multiple specimens and the advantage of an analysis chip that can obtain test results in a short time. Analytical devices have been proposed. Specifically, a plurality of microchannel structures are provided in a disk-shaped device, and a solid phase substance for reacting a substance contained in a sample is filled in a part of the channel. The measurement is performed by detecting the intensity of fluorescence obtained by exciting a label such as a fluorescent dye with a laser with a photodetector. Further, Patent Document 3 discloses a method using a structure of an optical disk by using a disk. A method is shown in which a biological sample or particles are attached to the same surface as the surface on which the track shape of an optical disk is formed, and a change in signal is detected using an optical pickup.

  In the methods of Patent Document 1 and Patent Document 2, the result of developing a sample in a flow path provided on a disk and reacting with a reagent is detected as a change in fluorescence intensity or absorbance, and the amount of antigen or antibody contained in the sample is determined. Measuring. The detection method is the same in principle as the method using a well plate, and the concentration of the labeling substance contained in the solution is measured by fluorescence or absorbance. Therefore, it is necessary to prepare a plurality of samples diluted in several stages for a sample having a narrow concentration range in which quantitative measurement is possible and a high concentration. On the other hand, as a method for detecting a low-concentration sample with high sensitivity, Patent Document 3 discloses a method for detecting a latex bead or magnetic bead on the focus surface of the optical disc. Detection of changes in output when the beads are placed at unspecified positions is insufficient, and they are easily affected by variations in the reflectance on the surface, dust, scratches, etc. There was a problem that it was difficult.

  The present invention has been made in view of such problems, and is an optical analysis capable of highly quantitative detection in a wide range from the low concentration to the high concentration of the antigen or antibody contained in the sample to be analyzed. An object is to provide an apparatus and an optical analysis method.

  In order to achieve the above object, an optical analyzer (10) according to a first aspect of the present invention comprises a first (1) configured by a groove (107) or a land (108) provided on a sample analysis disk (100). A pickup unit (12) for irradiating a track, and a second track and a third track adjacent to each other across the first track, and acquiring the reflected light by the pickup unit (12) and the pickup unit (12) The obtained reflected light detection unit (128) that simultaneously detects the reflected light from the first track, the second track, and the third track, and the first detected by the reflected light detection unit (128). The sample analysis disc is caused by fluctuations in reflected light in one track and fluctuations in reflected light in either the second track or the third track. (100) sample detection unit for detecting the quantity of bound biopolymers immobilized labeling beads (110) on the (18), characterized in that it comprises a.

  The optical analyzer (10) according to a second aspect of the present invention is the optical analyzer according to the first aspect, wherein the sample detector (18) is a reflected light fluctuation in the first track, and the reflected light detector (128). The material analysis is performed by fluctuations of reflected light in the second track or the third track adjacent to the first track in a direction opposite to the reading direction in the radial direction of the sample analysis disk (100) by The number of biopolymers bound to the labeling beads (110) immobilized on the disc (100) is detected.

  In the optical analyzer (10) according to a third aspect of the present invention, in the first or second aspect, the first track is a track constituted by the groove (107), the second track and the The third track is a track constituted by the land (108).

  According to a fourth aspect of the present invention, in the optical analyzer (10) according to the third aspect, the reflected light detector (128) reads the track on the second track or the third track. When the position is the center of the track and when the reading position on the track is shifted by a predetermined amount from the center of the track, the reflected light is detected, and the sample detector (18) detects the fluctuation of the reflected light at the center of the track and the track. The number of biopolymers bound to the labeling beads fixed on the material analysis disk (100) is detected by the fluctuation of the reflected light in a state shifted from the center by a predetermined amount.

  An optical analysis method according to a fifth aspect of the present invention includes a first track constituted by a groove (107) or a land (108) provided on a sample analysis disk (100), and the first track sandwiched between the first track and the first track. A reflected light detecting step of irradiating laser light to adjacent second and third tracks and simultaneously detecting reflected light from each track, in the first track detected by the reflected light detecting step A living body bound to the labeling beads (110) immobilized on the sample analysis disk (100) due to fluctuations in reflected light and fluctuations in reflected light in either the second track or the third track. A sample detection step of detecting the quantity of the polymer.

  According to the optical inspection apparatus of the present invention, detection can be performed by an apparatus having the same configuration as the optical pickup and the optical disk drive for reproducing information on the optical disk, and the apparatus can be reduced in size and cost. In addition, the labeling beads can be accurately counted at a high speed one by one without any restrictions on the position of the grooves or lands on the disk, so quantitative measurement from low to high concentration samples is possible. An apparatus can be provided.

Configuration block diagram of an optical inspection apparatus according to the present invention 1 is a conceptual diagram of a configuration of a pickup unit of an optical inspection apparatus according to the present invention. Schematic diagram of sample analysis disk configuration Cross-sectional schematic diagram of the track area in the sample analysis disk Schematic diagram showing the cross-sectional structure of the sample analysis disk and the reading state of the beads Schematic diagram showing sample reaction on sample analysis disk Schematic diagram showing light beam irradiation on a sample analysis disk in the optical inspection apparatus according to the present invention. The schematic diagram showing the reading state of the bead for label | marker fixed on the disk for sample analysis in the optical inspection apparatus which concerns on this invention Schematic diagram showing the immobilization position of labeling beads on the sample analysis disk The schematic diagram showing the reading state by the detrack of the labeling beads fixed on the sample analysis disk by the optical inspection apparatus according to the present invention The flowchart showing the flow of sample analysis in the optical inspection apparatus according to the present invention.

  First, the configuration of the optical inspection apparatus according to the present invention will be described with reference to FIGS. FIG. 1 is a block diagram showing the configuration of an optical inspection apparatus according to the present invention, and FIG. 2 is a diagram illustrating the configuration of a pickup section of the optical inspection apparatus according to the present invention.

  The optical inspection apparatus 10 includes a spindle motor 11, a pickup unit 12, a servo signal detection unit 13, a servo circuit 14, a main RF signal detection unit 15, an address detection unit 16, a sub RF signal detection unit 17, and a marker bead detection unit 18. A control unit 19 and a memory unit 20. The configuration of the reading device 1 may include other elements as necessary in addition to the above.

  2A, the pickup unit 12 includes an objective lens 121, a wave plate 122, a polarizing prism 123, a diffraction grating 124, a condenser lens 125, a laser oscillator 126, a detection lens 127, and a light detection unit 128. Is provided.

  In FIG. 1, a sample analysis disk 100 on which labeling beads 110 are arranged is installed so as to be capable of rotation control by a spindle motor 11, and rotates at a predetermined rotational speed. The sample analysis disc 100 has a disk shape with the same diameter as, for example, a CD (Compact Disc), a DVD (Digital Versatile Disc), and the like, and a similar material such as polycarbonate is used.

  Although the sample analysis disk 100 will be described separately, the sample dropped on the inner circumference by the centrifugal force due to rotation is developed in the outer circumference direction, and the labeling beads 110 by the antigen-antibody reaction are bound in the flow path, and the sample analysis is performed. A labeling bead 110 is fixed at a predetermined position of the disk 100 for use.

  A pickup section 12 that irradiates a predetermined spot of the sample analysis disk 100 with a light spot and detects the reflected light is installed so as to be movable in the radial direction of the sample analysis disk 100. The reflected light from the sample analysis disk 100 is converted into an electric signal by the light detection unit 128 in the pickup unit 12, and a part of the converted signal is sent to the servo signal detection unit 13.

  The servo signal detection unit 13 generates an FE (focus error) signal of the light spot irradiated on the sample analysis disk 100 from the signal from the pickup unit 12. In addition, a TE (tracking error) signal for the track on the sample analysis disk 100 is generated.

  The FE signal and the TE signal generated in the servo signal detection unit 13 are sent to the servo circuit 14 to control an electromagnetic actuator that drives the objective lens 121 in the pickup unit 12 in two directions, thereby performing an appropriate focus control for the light spot. And tracking control.

  Of the signals from the pickup unit 12, the signal from the first light receiving unit 130 </ b> A is sent to the main RF signal detection unit 15. The main RF signal detection unit 15 adds and amplifies four signals from the first light receiving unit 130A sent from the pickup unit 12 by a wideband amplifier to generate a main RF signal.

  The main RF signal generated by the main RF signal detection unit 15 is sent to the address detection unit 16. In the sample analysis disk 100, a track formed by a groove 107 and a land 108 to which a labeling bead 110 is fixed is formed in a spiral shape or a concentric shape. The address information indicating the track position is further provided in the sample analysis disk 100. For example, this address information can be set by cutting a part of a track and molding a pit having a plurality of lengths so that a modulation component corresponding to the address can be obtained, and can be taken out as a recognition signal for each track. It has become.

  The address detection unit 16 generates address information by detecting and demodulating a signal change caused by an address pit from the main RF signal. Further, the main RF signal is also sent to the labeling bead detector 18.

  Of the signals from the pickup unit 12, the signal from the second light receiving unit 130 </ b> B is sent to the sub RF signal detection unit 17. In the sub RF signal detection unit 17, the two signals from the second light receiving unit 130B sent from the pickup unit 12 are added and amplified by a wideband amplifier to generate a sub RF signal. The sub RF signal generated by the sub RF signal detection unit 17 is also sent to the labeling bead detection unit 18.

  The labeling bead detection unit 18 detects the labeling beads 110 fixed on the sample analysis disk 100 from the change in the main RF signal or the sub RF signal.

  The address information generated by the address detection unit 16 is sent to the control unit 19 as position information of the track counting the labeling beads 110. In addition, a signal indicating that the labeling beads 110 immobilized on the groove 107 or the land 108 are detected in the labeling bead detection unit 18 is also sent to the control unit 19 to count the number of labeling beads 110 in the currently measured track. Then, it is stored in the memory unit 20 together with the track information.

  The control unit 19 is configured by, for example, a CPU (Central Processing Unit) or a DSP (Digital Signal Processor). The control unit 10 performs various controls according to operations by an operation unit (not shown), various controls based on detection signals output from the light detection unit 9, and the like.

  Next, the configuration of the pickup unit 12 will be described with reference to FIG. A laser oscillator 126 is used as the light source. The laser oscillator 126 is configured by a laser diode. The wavelength of the laser light emitted from the laser oscillator 126 is appropriately selected depending on the size of the labeling bead 110. For example, when the labeling bead 110 having a diameter of about 140 nm is used, the wavelength of the laser light is It is desirable to use a laser diode that emits a blue-violet laser of about 400 nm. As the laser oscillator 126, for example, a semiconductor laser oscillator having the same wavelength of 405 nm as that for reproducing a Blu-ray (BD) disc can be used.

  The laser light emitted from the laser oscillator 126 becomes a parallel light beam by the condenser lens 125. Further, the light passes through the diffraction grating 124, is divided into three light beams, and passes through the polarizing prism 123.

  The laser light that has passed through the polarizing prism 123 is converted from linearly polarized light into circularly polarized light by passing through the wave plate 122, and the first reading spot 150 </ b> A and the first reading on the sample analysis disk 100 by the objective lens 121. Three reading spots consisting of two reading spots 150B and a third reading spot 150C are irradiated.

  An appropriate NA (Numerical aperture) of the objective lens 121 is selected depending on the size of the labeling beads 110. For example, when the labeling bead 110 having a diameter of about 140 nm is similarly used, it is desirable to use the objective lens 121 having an NA of about 0.85.

  In addition, the diffraction grating 124 is rotationally adjusted so that the first to third reading spots are appropriately irradiated onto the groove 107 and the land 108 on the sample analysis disk 100.

  The reflected light from the sample analysis disk 100 is linearly polarized light whose polarization plane is rotated by 90 degrees from the circularly polarized light when passing through the wave plate 122, is reflected by the polarizing prism 123, and passes through the detection lens 127. Then, the light detection unit 128 receives the light.

  The detection lens 127 has a mechanism for generating a focus error by a combination of a condenser lens and a semi-cylindrical lens. If the detection of the track or address pits on the sample analysis disk 100 and the labeling beads 110 cannot be detected with sufficient quality due to spherical aberration, another lens is added to the condenser lens 125 (not shown). It can also be improved by correcting spherical aberration.

  The light receiving unit 130 constituting the light detecting unit 128 has the configuration shown in FIG. 2B, and the reflected light of the first reading spot 150A is applied to the first light receiving unit 130A divided into four at the center. Then, the reflected light of the second reading spot 150B and the third reading spot 150C is applied to the second light receiving unit 130B and the third light receiving unit 130C, which are divided into two parts.

  The main RF signal detected by the main RF signal detection unit 15 and the sub RF signal detected by the sub RF signal detection unit 17 are generated by the sum of the divided cell outputs of the respective light receiving units constituting the light receiving unit 130, The FE signal and the TE signal are generated by calculation based on the divided cell output of each light receiving unit. This calculation is performed by the servo signal detection unit 13, and the calculation of the FE signal is generated by the calculation of (A + C) − (B + D) when expressed by the symbol of the divided cell in the light receiving unit 130, and the TE signal is [(A + D) − (B + C)] − k · [(E−F) + (G−H)] (k is a coefficient).

  Next, the structure of the sample analysis disk 100 read by the optical analyzer 10 of the present invention will be described with reference to FIGS.

  The sample analysis disk 100 is provided with a plurality of injection holes 101 on the inner peripheral side as a rotation target with respect to the center of the sample analysis disk 100. The injection hole 101 is a hole for dropping a sample, and its volume is sized to measure a sample required for inspection.

  In addition, flow paths 102 through which the dropped sample flows from each of the injection holes 101 are provided in a radial pattern when viewed from the center of the sample analysis disk 100. Furthermore, a detection region 104 is provided which is connected to each of the flow paths 102 and located on the outer periphery of the sample analysis disk 100. In addition, a part of the channel 102 is provided with a bead filling unit 103 filled with a predetermined amount of labeling beads 110 modified with an antibody that binds to a specific antigen contained in the sample.

  Further, a track region 105 having a spiral groove 107 having the same structure as the signal surface of the optical disc is provided on the outer periphery of the sample analysis disc 100 so as to include the detection region. Since the groove 107 is formed in the track region 105, the groove 107 and the land 108 serving as a groove exist on the reading surface of the sample analysis disk 100 as shown in FIG.

  FIG. 5 is a schematic diagram showing the cross-sectional structure of the sample analysis disk 100 and the reading state of the labeling beads 110. A flow path 102 having a depth of 100 μm to 500 μm is formed on the inner peripheral side of the sample analysis disk 100, and for labeling in which a specific antibody is modified in a part of the flow path 102 through which the sample to be analyzed passes. A bead filling unit 103 filled with beads 110 is provided.

  When the sample analysis disk 100 is rotated, the sample dropped from the injection hole 101 is guided to the flow path 102 for antigen-antibody reaction by centrifugal force. A bead filling unit 103 connected to the flow channel 102 is filled with a predetermined amount of labeling beads 110 modified with an antibody that binds to a specific antigen contained in the sample.

  As the labeling beads 110, polymer particles having a diameter of about 100 nm to 1 μm, or magnetic beads containing a magnetic material such as ferrite inside are used. It is also possible to use metal fine particles such as gold colloid or silica beads. The diameter of the labeling bead 110 is determined by the optical system of the reader 1 used for detection, and the optimum size for detection is determined. The reader 1 includes components used for existing optical disc drives, particularly optical pickups. There is an advantage that can be used.

  Among the optical disk readers currently in widespread use, the highest resolution is Blu-ray Disc (BD), which collects laser light with a wavelength of 405 nm on the disk with an objective lens with a numerical aperture (NA) of 0.85. Can be lighted. The shortest pit length of the disc signal is about 0.14 μm. Therefore, when the BD optical system is used, the signal can be detected as a signal up to the diameter of the labeling bead 110 of about 140 nm.

  An antibody 210 that specifically binds to the antigen 200 contained in the sample is bound to the surface of the labeling bead 110 in advance. There are various types of antibodies 210. For example, in the case of testing for hepatitis B, an HBsAg monoclonal antibody that specifically binds to an HBs antigen contained in blood is modified. The type of antibody 210 to be modified is selected to specifically bind to the type of antigen 200 to be detected.

  When the sample is mixed with the labeling bead 110 modified with the antibody 210 in the channel 102, the antigen 200 contained in the sample binds to the antibody 210 modified on the surface of the labeling bead 110, and the labeling bead 110. , A complex of antibody 210 and antigen 200 is formed.

  At this stage, depending on the amount of the antigen 200 contained in the sample, the labeling beads 110 of the complex in which the antigen 200 and the antibody 210 have reacted and the labeling beads 110 that have not reacted with the antigen 200 are constant. Present in proportion. Further, due to the centrifugal force of the sample analysis disk 100, the solution reacted in the flow channel 102 is developed in the detection region 104 in the outer peripheral portion.

  The detection area 104 has a spiral groove structure on the surface where the bottom surface of the flow path 102 contacts, like the signal surface of the optical disk.

  In these detection regions 104, an antibody 210 of the same type as that modified on the surface of the labeling bead 110 is fixed by a method such as silane coupling. The antigen 200 contained in the solution developed in the detection region 104, that is, the complex that has reacted with the labeling beads 110 modified with the antibody 210 in the flow channel 102 is further immobilized on the detection region 104. 210 is bound to form a sandwich-type complex of the labeling bead 110, the antibody 210, the antigen 200, and the immobilized antibody, and is immobilized on the substrate of the detection region 104. The labeling beads 110 that have not reacted with the antigen 200 are not limited to the detection area 104 but are further fed to the outer periphery of the disk by centrifugal force, and are discharged out of the detection area 104. A plurality of detection areas 104 are recorded with address information for specifying locations.

  Next, analysis of a sample using the sample analysis disk 100 will be described with reference to FIG.

  FIG. 6 is a diagram schematically showing the structure of the composite after the antigen 200 and the antibody 210 react on the sample analysis disk 100. This reaction is performed with a labeling bead 110, which is a measurable label, attached to the antigen 200 contained in the sample according to the principle of the sandwich method using the same antigen-antibody reaction as that performed in the well plate. It is fixed to the detection area 104. The sandwich method is often used as a method that facilitates quantitative measurement because an output is obtained linearly with respect to the absolute amount of a detection target. The immobilization method is not limited to this, and other methods such as a competition method can also be used.

  Next, the relationship between the track of the sample analysis disk 100 and the reading spot will be described. FIG. 7 shows the first reading spot 150A (B1), the second reading spot 150B (B2), and the third reading on the groove 107 (G) and the land 108 (L) forming the track of the sample analysis disk. It is the figure which showed a mode that the spot 150C (B3) was irradiated.

  Focus servo and tracking servo are performed on the disk track with reference to the first reading spot 150A arranged at the center. In the first reading spot 150A, the tracking can be performed using either the groove 107 as a reference or the land 108 as a reference. In this embodiment, the tracking is performed using the groove 107 as a reference. . However, the basics are the same when the land 108 is used as a reference.

  When tracking is performed using the groove 107 as a reference, when the tracking is adjusted with respect to the groove 107, the first reading spot 150A is positioned in the groove 107, and the second reading spot 150B and the third reading spot 150C. Is located on the adjacent land 108 across the first reading spot 150A.

  The measurement in this embodiment is usually performed for each track. Therefore, in the case of the sample analysis disk 100 in which concentric tracks are formed, the same track is traced as it is when tracking is applied. Therefore, when measuring the next track, the tracking servo is temporarily released and a kick pulse signal is input to jump to the next track.

  Even in this case, since the first reading spot 150A is controlled based on the groove 107, tracing is performed at positions B1 ′, B2 ′, and B3 ′ in FIG.

  When the track is formed in a spiral shape, the track sequentially moves, so that one track can be continuously traced by kicking back by adding a reverse kick pulse once during one rotation. .

  Here, when the labeling beads 110 are immobilized on the sample analysis disk 100 as shown in FIG. 8, each reading spot traces on each labeling bead 110. At this time, the outputs of the main RF signal (SB1) and the sub RF signals (SB2, SB3) change at the timing at which the labeling beads 110 are arranged as shown in FIG. When the output voltage of the main RF signal and the sub RF signal is equal to or lower than a predetermined threshold value, the labeling bead detection unit 18 counts the number of labeling beads 110 by counting by the labeling bead detection unit 18. Accurate measurement is possible.

  The size of each reading spot is determined by the wavelength of the laser beam from the laser oscillator 126 and the NA of the objective lens 121, and appropriately sets the relative relationship between the diameter of the labeling beads 110 to be used. For this reason, the change in the output voltage of the main RF signal and the sub RF signal can be sufficiently large with respect to the maximum value when the light is totally reflected and the minimum value where the reflected light is eliminated. The beads 110 can be accurately measured.

  In addition, the output from the second reading spot 150B and the third reading spot 150C can be obtained while tracing a specific track, but actually when jumping to the next track and reading it, Since the output from the third reading spot 150C and the output from the second reading spot 150B in the measurement after the track jump will trace the same land 108, the second reading spot 150B or the third reading spot Either one of 150C may be measured.

  Which track is counted is determined by the address detection unit 16, and the count number of the labeling beads 110 arranged in the groove 107 and the land 108 in a certain measurement track is accurately calculated by the control unit 19.

  When the labeling bead 110 is immobilized on the groove 107 and the land 108, the labeling bead 110 immobilized on the groove 107 is set so that the width of the groove 107 is slightly larger than the diameter of the labeling bead 110. Reading is performed in a state where the center of the labeling bead 110 substantially coincides with the center of the reading spot. However, when it is immobilized on the land 108, there is no restriction on the position of the labeling bead 110, and the center of the labeling bead 110 may be deviated from the center of the land 108.

  Since each reading spot is controlled by the tracking servo, each reading spot and the center of the land 108 are in the same state. FIG. 9 is a diagram showing the positional relationship between the groove 107 and the land 108 of the labeling bead 110 immobilized on the land 108.

  For example, as shown in FIG. 9, the labeling beads 110 fixed in the groove 107 are regulated by the width of the groove 107 and arranged at the center of the groove 107. However, the labeling beads 110 immobilized on the land 108 may be disposed at the center of the land 108 or may be disposed at a position shifted from the center.

  For this reason, when the center of the labeling bead 110 coincides with the center of the reading spot, the change of the RF signal is maximized, and the change is reduced according to the deviation from the center. For this reason, as shown in FIG. 9, the RF signal change is different when two labeling beads 110 on the land 108 are detected, and there is a possibility that a counting error occurs when counting is performed with a certain threshold as a reference.

  Therefore, a means for applying an arbitrary amount of offset voltage to the tracking servo of the reading spot and detracking the reading spot on the track with respect to the track center is provided. By performing detracking, a state in which the center of the reading spot coincides with the center of the labeling bead 110 fixed on the land 108 is detected, and the output signal is detected to maximize the amount of change in the RF signal. Can be.

  FIG. 10A is a conceptual diagram when a track composed of the groove 107 and the land 108 is viewed from above. For example, a labeling bead 110 is fixed on the land 108 at the position of Bead 2 which is the center of the land 108. In the state fixed in the position of Bead 1 shifted from the center of the land 108 to the upper side of the drawing, and in the state fixed to the position of Bead 3 shifted from the center of the land 108 to the lower side of the drawing, FIG. 10B shows an output state when the reading spot 150B is traced after being detracked in the plus direction (upward direction in the drawing) and in the minus direction (downward direction in the drawing).

  In FIG. 10B, in a normal state where no detracking is performed, the maximum amplitude is shown at the position of Bead2 centered on the land 108, and then the amplitude of Bead1 with the smallest deviation is followed by Bead3 with the largest deviation. Becomes smaller. If the threshold value at this time is set as shown by a two-dot broken line in FIG. 10B, only Bead2 is counted in the trace not to be detracked.

  If the threshold is set near the maximum output level and set to a value that counts all outputs, the possibility of erroneous counting increases due to fluctuations in the maximum output level, errors in reflectance, and the like.

  Further, in the detrack state in the plus direction, Bead1 shows the maximum amplitude, and then the amplitude decreases in the order of Bead2, and then Bead3. Conversely, in the detrack state in the negative direction, the amplitude decreases in the order of Bead3, Bead2, and Bead1. In this way, since the maximum amplitude can be obtained according to the position of the labeling bead 110 in the detrack state, the output and the detrack state are checked, and an accurate count can be performed by setting a threshold value.

  FIG. 11 is a flowchart showing the flow when counting the labeling beads 110. First, the tracking servo that traces a predetermined track is turned on (step S1). If the track has a spiral shape, a still pulse is added so that a predetermined track can be traced continuously.

  Next, the amplitude output 1 is measured in a normal state in which a predetermined track is not detracked (step S2). Next, detrack is generated in the + direction, and the amplitude output 2 is measured (step S3). Further, detrack is generated in the negative direction, and the amplitude output 3 is measured (step S4).

  Next, a threshold value is calculated and set from the measured amplitude outputs 1 to 3 (step S5). In addition, detrack amounts for the plus direction and the minus direction are also set. Next, an output that is equal to or lower than the threshold value in a normal state where no detracking is performed is counted to obtain a count number N1 (step S6). Next, in the set detrack + state, outputs that are similarly equal to or less than the threshold value are counted to obtain a count number N2 (step S7). Next, the number of outputs that are similarly equal to or less than the threshold in the set detrack state is counted to obtain a count number N3 (step S8). Finally, the obtained count numbers N1 to N3 can be added to obtain the number of labeling beads 110 immobilized on the land 108.

  As described above, the labeling beads 110 arranged on the track of the detection region using a plurality of reading spots can be accurately counted not only in the groove 107 but also on the land 108. This allows detailed quantification of the antibody or antigen contained in the biological sample. Also. By improving the accuracy of counting, it is possible to grasp low-concentration antibodies or antigens, and it is possible to expand the examination capability and application range.

  10: Optical analyzer, 11: Spindle motor, 12 pickup unit, 13: Servo signal detector, 14: Servo circuit, 15: Main RF signal detector, 16: Address detector, 17: Sub RF signal detector, 18: Labeling bead detection unit, 19: Control unit, 20: Memory unit, 121: Objective lens, 122: Wavelength plate, 123: Polarizing prism, 124: Diffraction grating, 125: Condensing lens, 126: Laser oscillator, 127 : Detection lens, 128: light detection unit, 130: light receiving unit, 130A: first light receiving unit, 130B: second light receiving unit, 130C: third light receiving unit, 200: antigen, 210: antibody

Claims (5)

Laser light is irradiated to a first track constituted by grooves or lands provided on a sample analysis disk, and a second track and a third track adjacent to each other across the first track. Pickup unit that acquires reflected light,
A reflected light detection unit that simultaneously detects reflected light from the first track, the second track, and the third track, acquired by the pickup unit;
Immobilized on the sample analysis disk by the fluctuation of the reflected light in the first track detected by the reflected light detector and the fluctuation of the reflected light in either the second track or the third track. A sample detector for detecting the number of biopolymers bound to the labeled beads,
An optical analyzer characterized by comprising: The sample detection unit is adjacent to the first track in a direction opposite to a variation in reflected light in the first track and a reading direction in the radial direction of the sample analysis disk by the reflected light detection unit. The number of biopolymers bound to the bead for label immobilized on the disk for analyzing the material is detected by fluctuation of reflected light in the second track or the third track,
The optical analyzer according to claim 1. The first track is a track configured by the groove, and the second track and the third track are tracks configured by the land,
The optical analyzer according to claim 1 or 2. The reflected light detection unit is configured such that the reading position on the track is the center of the track with respect to the second track or the third track, and the reading position on the track is shifted by a predetermined amount from the track center. Detecting the reflected light at
The sample detection unit has a living body bound to the bead for label immobilized on the disk for analyzing the material due to a change in reflected light at the center of the track and a change in reflected light in a state shifted from the center of the track by a predetermined amount. Detecting the quantity of macromolecules,
An optical analyzer according to claim 3. Laser light is irradiated to a first track constituted by grooves or lands provided on a sample analysis disk, and a second track and a third track adjacent to each other across the first track, A reflected light detection step for simultaneously detecting reflected light from the track;
Immobilization on the sample analysis disk by fluctuations in reflected light in the first track detected by the reflected light detection step and fluctuations in reflected light in either the second track or the third track. A sample detection step for detecting the number of biopolymers bound to the labeled beads,
An optical analysis method comprising the steps of:

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