JP2006349594A - Sample analysis disk and sample analyzer - Google Patents

Abstract

Disclosed is a sample analysis disk and a sample analysis apparatus capable of performing high-speed and accurate measurement with fewer samples than before.
A sample injection port (6) for injecting a sample, a flow path (7) provided inside the disk through which the sample flows, an overflow region (9) through which excess sample flows, and the sample introduced into the flow path. A recess 12 for fixing the beads 11 contained in the disk to the disk is provided on a side surface of the flow path 7, the sample is developed into the flow path 7 by centrifugal force generated by the rotation of the sample analysis disk 1, and the beads 11 are The sample can be analyzed at high speed from the light emission of the beads 11 contained in the sample by fixing to the recess 12.
[Selection] Figure 2

Description

  The present invention relates to a sample analysis disk for analyzing a sample and a sample analysis apparatus using the sample analysis disk.

  Gene expression has been studied to discover new therapeutic methods for diseases with various genetic associations in order to clarify gene function.

  In recent years, gene expression analysis techniques using semiconductor nanocrystals have been proposed. This semiconductor nanocrystal has a property of emitting light having a different wavelength depending on the composition and size of the semiconductor nanocrystal when irradiated with excitation light having a specific short wavelength (see, for example, Patent Document 1).

  Utilizing this property, a plurality of semiconductor nanocrystals with different emission wavelengths can be attached to plastic beads at an arbitrary ratio and encoded with a specific spectral distribution, and further, a specific mRNA is bound to each code bead. The probe and the mRNA are made to correspond one-to-one. In this way, the type of mRNA expressed can be analyzed by observing and decoding the spectral distribution of beads emitted by irradiating excitation light (see, for example, Patent Document 2).

  As a method for measuring fluorescent substances having different emission wavelengths, there is a fluorescence microscope apparatus that performs measurement using a plurality of bandpass filters having different pass bands (see, for example, Patent Document 3).

  FIG. 15 shows a conventional spectrum decoding apparatus that decodes the encoded beads from the spectrum distribution observed using the conventional fluorescence microscope apparatus. In FIG. 15, the conventional spectrum decoding apparatus includes an LED light source 40, a well plate 41 having a plurality of wells, a well plate driving means 42, an objective lens 43, an excitation light source 44, and bandpass filters 45a to 45. It comprises a pass filter 45h, a rotation filter 46, a rotation filter drive means 47, a CCD camera 48, a dichroic mirror 49, an image processing means 50, and a decoding means 51.

  The well plate 41 in which a sample containing a plurality of beads is injected into one well is moved by the well plate driving means 42 so that the well into which the sample has been injected becomes a measurement position. Then, the LED light source 40 is turned on from the top of the well plate 41, the bead silhouette is photographed by the CCD camera 48, and the objective lens 43 is moved to focus the bead. Then, the position of the bead is grasped from the silhouette image of the focused bead. This is because multiple beads injected into a well are not arranged, but they can be measured because the beads are attached to each other in a variety of states, such as being attached to each other or alone. This is done to locate the bead. Next, the beads are irradiated with excitation light from the excitation light source 44 using the dichroic mirror 49. The light of each bead emitted by the excitation light is photographed by the CCD camera 48 through the band-pass filter 45a to the band-pass filter 45h one by one by rotating the rotation filter 46 by the rotation filter driving means 47, and in the image processing means 50 Luminance spectrum data of each bead is obtained from the photographed image. Then, in the decoding means 51, the code of each bead is specified from the spectrum data of each bead.

  In addition, there is a sample analysis disk and a sample analysis device that develops a liquid sample such as blood or urine in a flow path inside the disk and optically scans and analyzes the sample (see, for example, Patent Document 4).

FIG. 16 shows a cross-sectional view of a conventional sample analysis disk. As shown in FIG. 16, the analysis disc has an injection part 60 that is a cylindrical cavity formed near the axis, a flow path 61 that communicates with the injection part 60 and extends to the outer periphery, and this flow path. An analysis region 62 installed in the middle of 61 and an absorption member 63 that communicates with the flow path 61 and is disposed at the outer peripheral end portion are provided inside. Then, the sample injected from the opening 64 opened on the upper surface of the disk is stored in the injection portion 60, and is moved to the analysis region 62 through the flow path 61 by centrifugal force when being mounted on the apparatus and rotated around the axis. The reagent is reacted with a reagent that reacts with the analysis target component in the sample placed in the analysis region 62, and the reaction state is optically detected by a detection means provided in the apparatus.
Special Table 2002-544488 Special table 2004-500109 gazette Japanese Patent Laid-Open No. 10-309281 JP 2003-215133 A

  However, the conventional configuration of the fluorescence microscope apparatus has a problem in that the cost is high because the excitation light source and the CCD camera are expensive.

  In the conventional configuration of the fluorescence microscope apparatus, when the distance between the beads is too close or in contact with each other, since each light emission affects each other, the beads must be excluded from the detection target. Therefore, there is a problem that it is not possible to detect all the beads within the photographing range.

  Therefore, since the conventional configuration of the fluorescence microscope apparatus cannot detect all the beads in the imaging range, it has a problem that the beads cannot be detected at high speed with a small amount of sample.

  In addition, the conventional configuration of the fluorescence microscope apparatus has a problem in that the apparatus itself cannot be reduced in size because each component of the optical system is not small.

  Further, the conventional configuration of the sample analysis disk has a problem that the beads cannot be detected because the beads cannot be fixed in the flow path.

  An object of the present invention is to solve the above-mentioned conventional problems, and to provide a sample analysis disk and a sample analysis apparatus that can detect each bead, increase the measurement speed, reduce the cost, and reduce the size. To do.

  In order to solve the above-described conventional problems, the sample analysis disk of the present invention is provided on the inner peripheral side of the disk, and is formed with an inlet through which the sample is injected, and from the inner peripheral side to the outer peripheral side of the disk. And a channel through which the sample flows, and the channel utilizes the centrifugal force acting from the inner periphery to the outer periphery in the radial direction of the disk by rotating the disk. A plurality of indentations for fixing a solid inspection object mixed in the side surface, and the indentations are formed to protrude from the side surface of the flow path to the outside of the flow path so that the objects are accommodated one by one. It is characterized by that.

  Further, in the sample analysis disk, an inlet provided on the inner circumference side of the disk and formed from the inner circumference side to the outer circumference side of the disk and having one end connected to the inlet and flowing through the sample. A passage having a width larger than the diameter of the solid inspection object mixed in the sample and smaller than twice the diameter of the inspection object, and the diameter of the inspection object. A plurality of sub-paths having a flow path length sufficiently shorter than the flow path length of the flow path having an outlet having a smaller width, and the inspection object rotates in the disk radial direction by rotating the disk; It is characterized in that it is fixed at the entrance of the secondary path using a centrifugal force acting from the inner periphery toward the outer periphery.

  Further, in the sample analysis disk, an inlet provided on the inner circumference side of the disk and formed from the inner circumference side to the outer circumference side of the disk and having one end connected to the inlet and flowing through the sample. The channel has a plurality of indentations for fixing the inspection object mixed in the sample on the bottom surface, and the indentations are provided below the bottom surface of the channel so that the objects can be accommodated one by one. It is formed to protrude to the side.

  Further, the sample analysis disk further includes an overflow region that is provided on the outer peripheral side of the disk and is connected to the other end of the flow path and into which excess sample flows.

  Further, in the sample analysis disk, the secondary path is disposed in contact with the side surface of the flow path.

  Further, in the sample analysis disk, the channel is a radial channel or a linear channel having a predetermined angle with respect to the radial direction.

  Further, the sample analysis disk is characterized by a disc shape formed with the same diameter as that of the CD or DVD.

  Furthermore, in the sample analysis disk, at least one sample can be measured, and at least one sample is formed for each sample.

  Further, in the sample analysis disk, a track pattern layer for performing tracking and focusing on the fixed inspection object is provided at the upper or lower portion of the flow path, and the track pattern layer is spiral or concentric. The track pattern is a groove or pit arranged.

  Further, in the sample analysis disk, data or a program can be recorded in the track pattern layer at the time of manufacturing the disk or additionally recorded at the time of measurement.

  Further, in the sample analysis disk, when the additional recording can be performed on the track of the track pattern layer, a land or a groove is provided, and when recording is possible at the time of manufacturing the disk, the track pattern track includes only a pit. It is a feature.

  Further, in the sample analysis disk, the test object is a bead or a magnetic bead.

  Further, in the sample analysis disk, the indentation has a rectangular parallelepiped shape.

  Further, in the sample analysis disk, the recess has a predetermined angle between the recess wall surface on the upstream side of the flow channel and the flow channel wall surface, or on both walls of the recess and the flow channel wall surface so that the test object can be easily accommodated. It is characterized by having.

  Further, in the sample analysis disk, a magnet is installed at the bottom of the recess so that the recess stably fixes the inspection object.

  Further, in the sample analyzer, a disk holding means for holding the sample analysis disk, a disk rotating means for rotating the sample analysis disk, a light emitting means for irradiating the sample analysis disk with light, and the sample analysis Detection means for detecting the inspection object fixed to the disk, servo control means for controlling the disk rotation means and the detection means, disk control means for controlling the light emitting means and the servo control means The test object injected into the sample analysis disk is measured.

  According to the sample analysis disk and sample analysis apparatus of the present invention, the apparatus can be made cheap and small, the beads can be detected one by one, the measurement efficiency is improved, and the number of samples is smaller than in the prior art. Can be measured at high speed.

  Embodiments of a sample analysis disk and a sample analyzer of the present invention will be described below in detail with reference to the drawings.

  FIG. 1 shows a cross-sectional view of a sample analysis disk in Example 1 of the present invention. In FIG. 1, a disk 1 for sample analysis is obtained by bonding a disk-shaped substrate 2 such as polycarbonate and a substrate 3 with bonding means (not shown) such as an ultraviolet curable adhesive or double-sided tape, A shaft hole 4 is provided at the center. On the lower surface of the substrate 2 to be bonded to the substrate 3, tracks such as pits or grooves are formed by processing means (not shown) such as injection molding, on which gold, silver or A reflective film made of a material such as aluminum is deposited, and a track pattern layer 5 coated with a protective film with good transmittance is formed on the reflective film to protect the reflective film and flatten the joint surface with the substrate 3. is doing. Further, in the substrate 3 disposed on the upper side, a sample inlet 6 in the thickness direction is provided in the vicinity of the shaft hole 4 at intervals along the circumferential direction, and grooves are formed on the substrate 2 side. A channel 7 is formed between the substrate 3 and the track pattern layer 5. Between the sample inlet 6 and the channel 7, a sample injection region 8 having a deeper groove than the channel 7 is provided in the substrate 3 in order to store the injected sample. In addition, an overflow region 9 having a deeper groove than the flow path 7 is provided in the substrate 3 in order to collect an excessive sample at the outer peripheral end of each flow path 7.

  FIG. 2 shows a front view of the sample analysis disk and an enlarged view of the flow path, and a positional relationship between the track pattern and the flow path. In FIG. 2, each flow path 7 is in a direction opposite to the rotation direction of the sample analysis disk 1 (in the case of FIG. 2, the rotation direction is clockwise) (counterclockwise in the case of FIG. 2), and for sample analysis. It is provided with a predetermined angle with respect to the radial direction of the disk 1 (any angle as long as it is smaller than 90 °. Here, for example, an angle of 35 °) is provided, and one flow path per sample. It consists of A sample including a plurality of beads is injected from the sample injection port 6 and collected in the sample injection region 8. Each channel 7 in the vicinity of the sample injection region 8 is provided with a capillary valve 10 having a larger space than the channel 7 in order to prevent sample movement due to capillary action during sample injection. The capillary valve 10 stops the movement of the sample due to capillary action by taking a large space with respect to the flow path 7. The sample moves in the flow path 7 by the centrifugal force acting in the outer peripheral direction of the sample analysis disk 1 by the rotation of the sample analysis disk 1.

  Each channel 7 is provided with a recess 12 on the side surface of the channel 7 for fixing the detection target bead 11 contained in the sample. Although the basic shape of the recess is a rectangular parallelepiped, the shape may be partially changed, for example, the bottom portion of the rectangular parallelepiped is circular so that the beads 11 are easily fixed like the recess 12. Even when the beads 11 are guided and fixed in the recess, the centrifugal force acting in the outer circumferential direction of the sample analysis disk 1 due to the rotation of the sample analysis disk 1 is used. The recess 12 has a depth direction distance that is longer than the diameter of the bead 11 (for example, 1.2 times the diameter of the bead 11) so that the bead 11 can easily fall, and the wall surface of the recess 12 and the flow path 7. The wall surface may have a predetermined angle (any angle as long as it is smaller than 90 °, for example, an angle of 35 °) for the beads 11 to fall into the recess 12 on the outer peripheral side wall surface of the flow path 7. The beads 11 are moved along the vicinity of the outer peripheral side wall surface of the flow path 7 by centrifugal force, and fallen and fixed one by one in the recess 12. Or you may provide a hollow in the reverse direction to the rotation direction of the disk of a flow path. The beads 11 used in the examples include magnetic beads. Therefore, when the bead 11 to be detected is a magnetic bead, the bead 11 can be more reliably fixed by installing a magnet at the bottom of each recess 12.

  The recess 12 is arranged so that the beads 11 fixed to the recess 12 are on the track pattern 70, but the track pattern 70 passing through the flow path 7 and the recess 12 is not arranged on the disk. Has a configuration.

  FIG. 3 shows a block diagram of a schematic configuration of a sample analyzer using the above-described sample analysis disk. In FIG. 3, the sample analyzer includes a disk holding means 20, a disk motor 21, a disk rotating means 22, a pickup 23, a rail 24 on which a dip-up moves, a light emitting means 25, a pickup motor 26, and a detection. It comprises means 27, servo control means 28, and disk control means 29. The servo control means 28 controls the disk rotation means 22, the pickup motor 26, and the detection means 27. Further, the disk control means 29 controls the light emitting means 25 and the servo control means 28.

  FIG. 4 shows the track information area of the sample analysis disk. In FIG. 4, the track pattern 70 is arranged on the disc in a spiral or concentric manner, and the track is formed of pits. The information area includes a data area 71 and a bead detection area 72. Addresses and data are recorded in the data area 71, and information such as addresses and servo control is recorded in the bead detection area 72, but no data is recorded. Note that the track information of the sample analysis disk 1 in the above-described embodiment includes only the bead detection region 72.

  FIG. 5 shows the format of each sector in each information area in the track of the sample analysis disk. In FIG. 5, there is a difference in the format between the data area 71 and the bead detection area 72. In the data area 71, a plurality of sectors 73 having the same length and storing address information and data are continuously arranged. ing. On the other hand, in the bead detection area 72, a sector 74 of the same length in which the address information and the bead detection notification information for notifying that a bead detection portion follows this sector is recorded flows in the rotation direction. One each is arranged in front of the road 7. A variable-length sector 75 in which a preamble for clock extraction and servo control and encoded random data are recorded is arranged in the track portion other than the above. The preamble of the sector 75 is for making it easy to re-acquire the clock because the track pattern 70 is interrupted by the flow path 7.

  Here, the depth of the pit is ¼ of the wavelength of the laser beam used. The focus error is detected by detecting a servo error signal from the detection means 27 using, for example, an SSD (Spot Size Detection) method. The tracking error is detected by detecting a servo error signal from the detection means 27 using, for example, a three beam system.

  A sample analysis process in this sample analyzer will be described. A sample including a plurality of beads is injected into the sample analysis disk 1 in advance from the sample injection port 6 into the sample injection region 8, and the sample analysis disk 1 is transferred to a disk motor 21 by a disk holding means 20 such as a magnet. It is fixed. The sample in the sample injection region 8 is prevented from moving to the flow path 7 by the capillary valve 10 before starting the analysis.

  In this state, when analysis is started, the disk control means 29 sends a command to the disk rotation means 22 via the servo control means 28 in order to develop the sample in the sample injection region 8 into the flow path 7. Controls the disk motor 21 so that the sample analysis disk 1 is rotated by the disk motor 21 at a predetermined time and a predetermined rotation speed (for example, several minutes, 6,000 rotations / minute). The sample starts to move to the flow path 7 by the centrifugal force acting in the outer circumferential direction of the sample analysis disk 1 due to the rotation of the sample analysis disk 1. While the detection target beads 11 included in the sample are moving in the flow path 7, one bead 11 falls and is fixed to one recess 12 provided on the side surface of the flow path 7 on the outer peripheral side of the disk by centrifugal force.

  During the development of the sample in the flow path 7, the pickup 23 traces the track pattern 70 in the overflow area 9. At this time, the disk motor 21 is controlled by the disk rotating means 22 so that the linear velocity of the trace portion is constant. The space in the overflow region 9 before the sample is developed is filled with air, but when the sample development is completed, an excess sample is accumulated in the space in the overflow region 9. The difference between before and after the sample is developed is detected by the detecting means 27, and it is detected that the sample has been developed into the flow path 7. Alternatively, a method of applying a reagent that reacts with the liquid to the bottom surface of the overflow region 9 and detecting the reaction between the liquid and the reagent may be used.

  When the development of the sample in the flow path 7 is completed, the disk motor 21 next detects the bead 11 fixed to the recess 12 at the irradiation point of the laser beam irradiated from the pickup 23 to the sample analysis disk 1. Is controlled by the disk rotating means 22 so that the linear velocity is constant. Here, the laser light emitted from the pickup 23 is light having a short wavelength capable of exciting the beads 11. Servo detection and bead excitation are performed using laser light having a short wavelength. The pickup 23 moves on the rail 24 by the pickup motor 26, traces the track pattern 70 in the track pattern layer 5 of the sample analysis disk 1, and has a short wavelength that can excite the beads 11 on the sample analysis disk 1. Irradiate the laser beam. A part of the irradiated laser light is reflected on the track pattern layer 5, and the reflected light is detected by the detecting means 27 as servo control information. This detection information is sent to the disk control means 29, and the disk control means 29 has a best focus (for example, a spot diameter equal to or smaller than the diameter of the beads 11) with respect to the beads 11 fixed along the track. The light emitting means 25 is controlled so as to maintain the defocused state in the track pattern layer 5 so that the position at a predetermined distance from the track pattern layer 5 is in focus. Then, the disc control means 29 uses the detected information so that the laser light irradiated from the pickup 23 to the sample analysis disc 1 traces the track pattern 70, and the linear velocity is constant from the information detected from the track. Then, an instruction is sent to the servo control means 28. The servo control means 28 controls the disk rotation means 22, the pickup motor 26 and the detection means 27 in accordance with instructions from the disk control means 29.

  The detection means 27 optically detects the light emission from the beads 11 irradiated with the excitation light, and sends the obtained information to the disk control means 29. Since each bead 11 is coded with a specific spectral distribution for light emission, the detection means 27 detects the light emission from the bead 11 as luminance information for each predetermined spectrum and sends it to the disk control means 29 as spectral distribution data. . The spectrum distribution data of each bead 11 sent from the detection means 27 to the disk control means 29 is identified by the code of each bead 11 by a comparison operation unit (not shown) or an external comparison operation unit (not shown). Is done.

  In the above-described embodiment, one channel 7 is used for one sample, but a plurality of channels may be used for one sample. Thereby, it is possible to prevent dust or the like mixed in the sample from clogging the flow path 7 and failing in the analysis.

  Further, if the outer shape of the sample analysis disk 1 is equivalent to that of an existing optical disk medium, such as a CD or DVD, the existing optical disk manufacturing stamper, disk rotating means, and feeding mechanism can be shared, and the analyzer Can be configured at a low price.

  FIG. 6 shows a cross-sectional view of the sample analysis disk when the track pattern layer 5 is above the flow path 7. As shown in FIG. 6, a sample analysis disk 1 having a structure in which the track pattern layer 5 is provided on the upper part of the flow path 7 may be used. 1 and FIG. 6, when the laser beam having the same output is irradiated to the beads 11 fixed to the recess 12, the track pattern layer 5 in the disk structure of FIG. Is not attenuated, a strong emission intensity is obtained from the beads 11 and detection with higher accuracy is possible.

  If a program or data is recorded in the data area 71 of the track pattern layer 5 in the format shown above at the time of manufacturing the disk, parameters and programs required at the time of analysis are read from the disk at the start of analysis, so that A part of the program pattern describing a series of processes of sample analysis stored in is unnecessary, and the apparatus can be simplified.

  However, since it is limited to use as a ROM disk, it is not convenient for users. In addition, the manufacturing side must manufacture a disc for each type of inspection, which is inefficient. Therefore, if a disc structure having a recordable track pattern layer in which an organic dye or the like is applied between the polycarbonate substrate and the reflective layer is used, the optimum measurement sequence program and parameters for the sample to be measured can be obtained by the user. Since it can write in the data area 71 before measurement, it can obtain a great effect as a highly versatile disc.

  Here, the track pattern 70 of the recordable recordable disc is composed of lands or grooves. The format of each sector in the information area and each information area is the same as that of the ROM disk described above. However, the address information, the bead detection notification information, and the servo control signal are obtained by periodically modulating the groove edge and obtaining the signal therefrom. The tracking error is detected by detecting the servo error signal from the detecting means 27 using, for example, a push-pull method. Note that all tracks of the recordable disk need not be composed of lands or grooves. For example, the track pattern 70 in the data area 71 may be composed of lands or grooves, and the track pattern 70 in the bead detection area 72 may be composed of pits.

  FIG. 7 shows a front view of a sample analysis disk and an enlarged view of a channel in Example 2 of the present invention, and a positional relationship between a track pattern and a channel. In FIG. 7, each flow path 7 is provided along the radial direction of the sample analysis disk 13, and is composed of one flow path per sample. A sample including a plurality of beads is injected from the sample injection port 6 and collected in the sample injection region 8. Each channel 7 in the vicinity of the sample injection region 8 is provided with a capillary valve 10 having a larger space than the channel 7 in order to prevent sample movement due to capillary action during sample injection. The sample moves in the flow path 7 by the centrifugal force acting in the outer peripheral direction of the sample analysis disk 13 by the rotation of the sample analysis disk 13. The difference from the configuration of Example 1 is that the main path 14 having a width sufficiently wider than the diameter of the beads 11 (for example, twice or more the diameter of the beads 11) and the entrance are wider than the diameter of the beads 11 in the flow path 7. A secondary path having a width (for example, 1.2 times the diameter of the bead 11) and a width smaller than the diameter of the bead 11 (for example, 0.7 times the diameter of the bead 11) capable of fixing the bead 11 at the outlet. 15 are each a channel length of a predetermined length sufficiently shorter than the channel length of the channel 7 (for example, 1 to 2 times the diameter of the bead 11) and parallel to the width direction of the channel 7 It is a point provided.

  Similar to the first embodiment, the track pattern 70 of the track pattern layer 5 is spirally or concentrically arranged on the disk, and the track is formed of pits. The track configuration and format are the same as in the first embodiment. The sub-path 15 is arranged so that the beads 11 fixed to the sub-path 15 are on the track pattern 70, but the track pattern 70 of the portion that passes through the flow path 7 is not written on the disk. I am doing.

  A sample analysis process in this sample analyzer will be described. The sample analysis process in the sample analyzer differs from the sample analysis process of the first embodiment in that the beads 11 contained in the sample moving through the flow path 7 are fixed to the sample analysis disk 13. .

  Similar to the sample analysis process of the first embodiment, a sample containing a plurality of beads is previously injected into the sample analysis disk 13 and is fixed to the disk motor 21 by the disk holding means 20 such as a magnet. When the analysis is started, the disk control means 29 sends a command to the disk rotation means 22 via the servo control means 28 in order to develop the sample in the sample injection region 8 into the flow path 7, and the disk rotation means 22 The disk motor 21 is controlled so that the analysis disk 13 is rotated by the disk motor 21 at a predetermined time and a predetermined rotation speed (for example, several minutes, 6,000 rotations / minute). The sample starts to move to the flow path 7 by the centrifugal force acting in the outer circumferential direction of the sample analysis disk 13 due to the rotation of the sample analysis disk 13. The bead 11 to be detected contained in the sample passes through either the main path 14 or the sub-path 15 provided in the flow path 7 in parallel with the width direction of the flow path 7 while moving in the flow path 7. try to. Here, the beads 11 cannot pass through the sub-path 15 and are fixed, but the main path 14 has a width that is sufficiently wider than the diameter of the beads 11, so that the beads 11 can pass through the main path 14. The sub-path 15 is provided so that the beads 11 fixed to the sub-path 15 are on the track of the sample analysis disk 13, and the distance between the tracks is sufficiently wider than the bead diameter (for example, the beads 11 4 times the diameter).

  Since the detection of the completion of the development of the sample in the flow path 7 and the subsequent processes are the same as those in the first embodiment, description thereof will be omitted.

  FIG. 8 shows a front view of the sample analysis disk and an enlarged view of the channel, and a positional relationship between the track pattern and the channel. In FIG. 8, each flow path 7 is in the opposite direction (counterclockwise in FIG. 8) to the rotation direction of the sample analysis disk 13 (in the case of FIG. 8, the rotation direction is clockwise), and for sample analysis. It is provided with a predetermined angle with respect to the radial direction of the disk 13 (any angle as long as it is smaller than 90 °. Here, for example, an angle of 35 °) is provided, and one flow path per sample. It consists of In this way, the flow path 7 has a predetermined angle with respect to the radial direction of the sample analysis disk 13 (any angle may be used as long as it is an angle smaller than 90 °, for example, an angle of 35 ° here). In the case of a straight flow path or a curved flow path such as an arc or a spiral, in order to efficiently use the centrifugal force acting in the outer peripheral direction of the sample analysis disk 13 due to the rotation of the sample analysis disk 13 for fixing the beads 11. The sub-path 15 is provided in the flow path side area in the disk outer peripheral direction. By arranging the sub-path 15 as described above, the beads 11 can be easily guided and fixed to the sub-path 15.

  In the second embodiment, similarly to the first embodiment, a single sample may be configured with a plurality of flow paths. Thereby, it is possible to prevent dust or the like mixed in the sample from clogging the flow path 7 and failing in the analysis. Similarly to the first embodiment, if the outer shape of the sample analysis disk 13 is the same as that of an existing optical disk medium such as a CD or DVD, the existing optical disk manufacturing stamper, disk rotating means, and feeding mechanism are shared. The analyzer can be configured at a low price.

  FIG. 9 shows a cross-sectional view of the sample analysis disk when the track pattern layer 5 is located above the flow path 7. Similarly to the first embodiment, a sample analysis disk 13 having a structure in which the track pattern layer 5 as shown in FIG. Thus, in the disk structure of FIG. 9, the track pattern layer 5 does not attenuate the amount of laser light, so that a strong emission intensity can be obtained from the beads 11 and detection with higher accuracy is possible.

  If a program or data is recorded in the data area 71 of the track pattern layer 5 in the same format as described in the first embodiment at the time of manufacturing the disc as in the first embodiment, parameters and programs necessary for the analysis at the start of the analysis are stored. By reading from the disk, a part of the program pattern describing a series of processes of sample analysis stored in the sample analyzer becomes unnecessary, and the apparatus can be simplified.

  In addition, if the disc structure has a track pattern layer that can be additionally recorded and coated with an organic dye between the polycarbonate substrate and the reflective layer, the program and parameters of the measurement sequence that are optimal for the sample to be measured can be set by the user. Since it can write in the data area 71 before measurement, it can obtain a great effect as a highly versatile disc.

  Here, the track of the recordable disc is composed of lands or grooves, the details of which are the same as in the first embodiment.

  FIG. 10 shows a cross-sectional view of a sample analysis disk in Example 3 of the present invention. In FIG. 10, the difference from the configuration of the first embodiment is that the flow path 7 forms a substrate 76 provided with a recess 17 for fixing the detection target beads 11 included in the sample in the sample analysis disk 16. This is a point provided between the substrate 2 and the substrate 3 to be processed.

  FIG. 11 shows a front view of the sample analysis disk and an enlarged view of the flow path, and a positional relationship between the track pattern and the flow path. In FIG. 11, each flow path 7 is provided along the radial direction of the sample analysis disk 16, and is composed of one flow path per sample. A sample including a plurality of beads is injected from the sample injection port 6 and collected in the sample injection region 8. Each channel 7 in the vicinity of the sample injection region 8 is provided with a capillary valve 10 having a larger space than the channel 7 in order to prevent sample movement due to capillary action during sample injection. The sample moves in the flow path 7 by centrifugal force and capillary action acting in the outer peripheral direction of the sample analysis disk 16 by the rotation of the sample analysis disk 16. The shape of the recess 17 is a rectangular parallelepiped, and the size of the recess 17 is such that one bead is inserted (for example, vertical and horizontal are 1.1 times the diameter of the bead 11 and the depth is 1.2 times the diameter of the bead 11). ).

  Similar to the first embodiment, the track pattern 70 of the track pattern layer 5 is spirally or concentrically arranged on the disk, and the track is formed of pits. The track configuration and format are the same as in the first embodiment.

  The recess 17 is arranged so as to be on the track pattern 70, but the track pattern 70 in the portion passing through the flow path 7 is not written on the disk.

  FIG. 12 shows a block diagram of a schematic configuration of a sample analyzer using the above-described sample analysis disk. In FIG. 12, the sample analyzer includes a disk holding means 20, a disk motor 21, a disk rotating means 32, a pickup 23, a rail 24 on which a dip-up moves, a light emitting means 25, a pickup motor 26, and a detection. It comprises means 27, servo control means 30, and disk control means 31. The servo control means 30 controls the disk rotation means 32, the pickup motor 26, and the detection means 27. The disk control means 31 controls the light emitting means 25 and the servo control means 30.

  A sample analysis process in this sample analyzer will be described. A sample is previously injected into the sample analysis disk 16 from the sample injection port 6 into the sample injection region 8, and the sample analysis disk 16 is fixed to a disk motor 21 by disk holding means 20 such as a magnet. The sample in the sample injection region 8 is prevented from moving to the flow path 7 by the capillary valve 10 before starting the analysis.

  When analysis is started in this state, the disk control means 31 sends a command to the disk rotation means 32 via the servo control means 30 in order to develop the sample in the sample injection region 8 into the flow path 7. The disk rotating means 32 controls the disk motor 21 so that the sample analysis disk 16 is rotated by the disk motor 21 at a predetermined time and a predetermined rotation speed (for example, 6,000 rotations / minute for several tens of seconds). The capillary passes through the capillary valve 10 by the centrifugal force acting in the outer circumferential direction of the sample analysis disk 16 by the rotation of the sample analysis disk 16. Thereafter, the disk rotating means 32 is rotated at a rotational speed at which the beads 11 can fall into the depressions 17 by gravity by the disk motor 21 in response to a command from the disk control means 31 via the servo control means 30 (for example, , 60 rotations / minute), the disk motor 21 is controlled to rotate for a predetermined time (for example, several tens of minutes). The sample is developed into the flow path 7 by capillary action and centrifugal force acting in the outer peripheral direction of the sample analysis disk 16. The beads 11 to be detected contained in the sample are moved in the flow path 7, and one bead is provided for each depression 17 provided on the bottom surface of the flow path 7 due to gravity acting toward the bottom surface of the sample analysis disk 16. 11 is depressed and fixed. The depression 17 is arranged so that the beads 11 are fixed on the track of the track pattern layer 5. The beads used in the examples include magnetic beads. Therefore, when the detection target bead 11 is a magnetic bead, the bead 11 can be more reliably fixed by installing a magnet at the bottom of each recess 17. Alternatively, the magnetized ferromagnetic material 33 may be provided below the sample analysis disk 16 while the sample is being developed in the flow path 7. As a result, the beads 11 are attracted toward the bottom surface of the disk by gravity and magnetic force, and can be easily dropped into the recess 17.

  Since the detection of the completion of the development of the sample in the flow path 7 and the subsequent processes are the same as those in the first embodiment, description thereof will be omitted.

  In the third embodiment, similarly to the first embodiment, a plurality of flow paths may be configured per sample. Thereby, it is possible to prevent dust or the like mixed in the sample from clogging the flow path 7 and failing in the analysis.

  Similarly to the first embodiment, if the outer shape of the sample analysis disk 16 is the same as that of an existing optical disk medium, such as a CD or DVD, the existing optical disk manufacturing stamper, disk rotating means, and feeding mechanism are shared. The analyzer can be configured at a low price.

  FIG. 13 shows an enlarged view and a cross-sectional view of the flow path of the sample analysis disk. In FIG. 13, (a) is the flow path 7 in the embodiment 3 described so far in which the bottom face of the flow path 7 is flat, and (b) is inclined toward the recess 17 at the bottom face of the flow path 7. . Here, as shown in (b), the bottom surface of the flow channel 7 may be a flow channel having an inclination toward the recess 17. This makes it easier for the beads 11 to fall into the recess 17 than in the flow path 7 of FIG.

  FIG. 14 is a cross-sectional view of the sample analysis disk when the track pattern layer 5 is above the flow path 7. Similarly to the first embodiment, a sample analysis disk 16 having a structure in which the track pattern layer 5 as shown in FIG. Thereby, in the disk structure of FIG. 14, the light quantity of the laser light is not attenuated by the track pattern layer 5, so that a strong emission intensity can be obtained from the beads 11, and detection with higher accuracy is possible.

  If a program or data is recorded in the data area 71 of the track pattern layer 5 in the same format described in the first embodiment at the time of manufacturing the disc as in the first embodiment, the parameters and programs necessary for the analysis at the start of the analysis are stored. By reading from the disk, a part of the program pattern describing a series of processes of sample analysis stored in the sample analyzer becomes unnecessary, and the apparatus can be simplified.

  In addition, if the disc structure has a track pattern layer that can be additionally recorded and coated with an organic dye between the polycarbonate substrate and the reflective layer, the program and parameters of the measurement sequence that are optimal for the sample to be measured can be set by the user. Since it can write in the data area 71 before measurement, it can obtain a great effect as a highly versatile disc.

  Here, the track of the recordable disc is composed of lands or grooves, the details of which are the same as in the first embodiment.

  The sample analysis disk and sample analysis apparatus according to the present invention are useful for bioanalysis apparatuses and the like, and are particularly suitable for use as gene expression analysis apparatuses.

Sectional drawing of the disk for sample analysis in Example 1 of this invention The front view of the sample analysis disk in Example 1 of this invention, the enlarged view of a flow path, and the figure showing the positional relationship of a track pattern and a flow path 1 is a block diagram of a sample analyzer in Embodiment 1 of the present invention. The figure showing the information area of the track | truck of the sample analysis disk in Example 1 of this invention The figure showing the format of each sector in each information area in the track | truck of the disk for sample analysis in Example 1 of this invention. Sectional drawing of the disk for sample analysis in Example 1 of this invention The front view of the sample analysis disk in Example 2 of this invention, the enlarged view of a flow path, and the figure showing the positional relationship of a track pattern and a flow path The front view of the sample analysis disk in Example 2 of this invention, the enlarged view of a flow path, and the figure showing the positional relationship of a track pattern and a flow path Sectional drawing of the disk for sample analysis in Example 2 of this invention Sectional drawing of the disk for sample analysis in Example 3 of this invention The front view of the sample analysis disk in Example 3 of this invention, the enlarged view of a flow path, and the figure showing the positional relationship of a track pattern and a flow path Block diagram of a sample analyzer in Embodiment 3 of the present invention The enlarged view of the flow path of the sample analysis disk in Example 3 of this invention, and sectional drawing of a flow path Sectional drawing of the disk for sample analysis in Example 3 of this invention Block diagram of a coding device with conventional spectrum Sectional view of a conventional sample analysis disk

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Sample analysis disk 2 Substrate 3 Substrate 4 Shaft hole 5 Track pattern layer 6 Sample injection port 7 Flow path 8 Sample injection area 9 Overflow area 10 Capillary valve 11 Detection target beads 12 Indentation 13 Sample analysis disk 14 Main path 15 Sub path 16 Sample analysis disk 17 Recess 20 Disk holding means 21 Disk motor 22 Disk rotating means 23 Pickup 24 Rail 25 Light emitting means 26 Pickup moving motor 27 Detection means 28 Servo control means 29 Disk control means 30 Servo control means 31 Disk control means 32 disc rotating means
DESCRIPTION OF SYMBOLS 33 Ferromagnetic material 40 LED light source 41 Well plate 42 Well plate drive means 43 Objective lens 44 Excitation light source 45a-45h Band pass filter 46 Rotation filter 47 Rotation filter drive means 48 CCD camera 49 Dichroic mirror 50 Image processing means 51 Decoding means 60 Injection Section 61 Flow path 62 Analysis area 63 Absorbing member 64 Opening 70 Track pattern 71 Data area 72 Bead detection area 73 Sector 74 Sector 75 Sector 76 Substrate

Claims (16)

An inlet provided on the inner circumferential side of the disc, into which the sample is injected,
Formed from the inner peripheral side of the disc to the outer peripheral side, with one end connected to the inlet and a flow path through which the sample flows,
The flow path has a plurality of indentations that fix a solid test object mixed in the sample by utilizing a centrifugal force that works from an inner periphery to an outer periphery in the radial direction of the disk by rotating the disk. Have
The indentation is formed to protrude from the side surface of the flow path to the outside of the flow path so that the objects are accommodated one by one.
A sample analysis disc characterized by the above. An inlet provided on the inner circumferential side of the disc, into which the sample is injected,
Formed from the inner peripheral side of the disc to the outer peripheral side, with one end connected to the inlet and a flow path through which the sample flows,
The flow path has an inlet having a width larger than the diameter of the solid inspection object mixed in the sample and smaller than twice the diameter of the inspection object, and an outlet having a width smaller than the diameter of the inspection object. Having a plurality of sub-channels with a channel length sufficiently shorter than the channel length of the channel,
The inspection object is fixed to the entrance of the sub-path using a centrifugal force that works from the inner periphery to the outer periphery in the radial direction of the disk by rotating the disk.
A sample analysis disc characterized by the above. An inlet provided on the inner circumferential side of the disc, into which the sample is injected,
Formed from the inner peripheral side of the disc to the outer peripheral side, with one end connected to the inlet and a flow path through which the sample flows,
The flow path has a plurality of depressions on the bottom surface for fixing an inspection object mixed in the sample,
The indentation is formed to protrude downward from the bottom surface of the flow path so that the objects are accommodated one by one.
A sample analysis disc characterized by the above. Provided on the outer peripheral side of the disk, further comprising an overflow region connected to the other end of the flow path and into which excess sample flows.
4. The sample analysis disk according to claim 1, wherein the sample analysis disk is a disk. 3. The sample analysis disk according to claim 2, wherein the sub-path is disposed in contact with the side surface of the flow path. 4. The sample analysis disk according to claim 1, wherein the flow path is a radial flow path or a straight flow path having a predetermined angle with respect to the radial direction. 5. 4. The sample analysis disk according to claim 1, wherein the sample analysis disk has a disk shape having the same diameter as that of a CD or DVD. 4. The sample analysis disk according to claim 1, wherein at least one sample is measurable, and at least one sample is formed for each sample. 5. A track pattern layer for performing tracking and focusing on the fixed inspection object is provided at the upper or lower portion of the flow path, and the track pattern layer is a groove or pit arranged in a spiral or concentric shape. 4. The sample analysis disk according to claim 1, further comprising a certain track pattern. 10. The sample analysis disk according to claim 9, wherein data or a program can be recorded on the track pattern layer at the time of manufacturing the disk or additionally recorded at the time of measurement. 11. The track of the track pattern layer includes a land or a groove when the additional recording is possible, and the track of the track pattern includes a pit only when recording is possible at the time of manufacturing the disc. Disc for sample analysis as described. 4. The sample analysis disk according to claim 1, wherein the inspection object is a bead or a magnetic bead. 4. The sample analysis disk according to claim 1, wherein the indentation has a rectangular parallelepiped shape. The indentation is characterized in that the indentation wall surface and the inflow channel wall surface on the upstream side of the channel or the indentation wall surfaces and the inflow channel wall have a predetermined angle so that the inspection object is easily accommodated. The disk for sample analysis according to claim 13. The sample analysis disk according to claim 12, wherein a magnet is installed at the bottom of the recess so that the recess stably fixes the inspection object. A disk holding means for holding the sample analysis disk according to any one of claims 1 to 3, a disk rotating means for rotating the sample analysis disk, and light for irradiating the sample analysis disk with light. An emission means, a detection means for detecting the inspection object fixed to the sample analysis disk, a servo control means for controlling the disk rotation means and the detection means, the light emission means and the servo control means. A sample analysis apparatus comprising: a disk control means for controlling, and measuring an inspection object injected into the sample analysis disk.

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