JP2006053050A - Inspection method of particle for inspection, and inspection device of particle for inspection - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an inspection device capable of detecting accurately each relative order position of fine particles arrayed on a plate substrate. <P>SOLUTION: The particles 19 are inserted into a passage 14 provided on the plate substrate 12. The particles 19 comprise a particle 19a for inspection having a probe attached to its surface and a reference point particle 19b for setting an inspection start point. A relative moving distance of laser light 24 is reset at the point of time when the reference point particle 19b is detected, and the relative moving distance L of the laser light 24 is measured by using the position of the reference point particle 19b as an inspection start point, to thereby suppress deviation between the estimated position of the particle 19a for inspection from the reference point particle 19b and the actual position by suppressing accumulation of tolerances. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a simple inspection fine particle inspection method and inspection fine particle inspection apparatus capable of performing, for example, a blood test, a urine test, or a DNA test in a medical institution or an individual. The present invention relates to an inspection fine particle inspection method and an inspection fine particle inspection apparatus capable of accurately calculating the position from the reference point.

  2. Description of the Related Art In recent years, development of chips for testing specimens collected from the human body, such as blood and urine, has become active. For example, a DNA chip is a product in which many types of DNA fragments (referred to as probes or reagents) are pasted on a substrate such as glass, and a specific gene from genes collected from humans (referred to as specimens or targets). Multiple types of cancer genes (such as oncogenes) can be detected at a time.

  Conventionally, the detection of biomolecules, which has been performed with a test tube and a dropper, or a stirrer, can be performed at a high speed, and the inspection process can be simplified. It is considered and attracts attention.

  By the way, test chips are currently mainly developed as research chips for universities and research institutions, but in the future, simple test chips for medical institutions and individuals will be commercialized. There is expected.

  The inspection chip is composed of, for example, a plate substrate (inspection base) having a channel formed in a groove shape, and a lid body that is superimposed on the plate substrate and joined to the plate substrate.

  In the inspection method, a probe having a specific base sequence is arranged in the flow path, and a label such as a fluorescent dye is added to DNA (specimen) collected from a human body in the flow path. When DNA having a base sequence complementary to the probe is present in the sample, it is captured by a hybridization reaction with the probe.

  For example, the probe can be distinguished by a fluorescent dye having a different emission wavelength from the fluorescent dye added to the specimen, and is included in the specimen by detecting which probe has undergone a hybridization reaction with the DNA. The base sequence of DNA can be specified.

  FIG. 5 is a plan view of the plate substrate 2 in which a plurality of fine particles 1 are arranged in a row in the groove 2a. The groove 2a is a flow path through which a solution containing the specimen flows, and an inflow port 2b is formed upstream of the flow path, and an outflow port 2c is formed in a concave shape on the downstream side. A solution containing a sample such as DNA collected from a person is injected from the inflow port 2b and flows into the groove 2a. At this time, if DNA complementary to the probe fixed to the microparticle 1 is present in the specimen, the probe and DNA are fixed by causing a hybridization reaction. If the fluorescence intensity is measured for each fine particle 1, it can be easily determined whether or not DNA has been captured.

  FIG. 6 is a conceptual diagram showing a conventional fluorescence detection apparatus. The probe is fixed to fine particles 1 made of resin, glass or the like and having a diameter of about 100 μm. The fine particles 1 are arranged inside grooves 2 a formed in the plate 2. Laser light 4 emitted from the laser light source 3 is irradiated onto the fine particles 1 on the plate 2 through the mirror 5 and the lens 6 to excite the fluorescent dye for specimen labeling hybridized with the fluorescent dye or the probe in the fine particles 1. The excited fluorescent dye emits fluorescence R having a wavelength characteristic of the fluorescent dye, and this fluorescence R is detected by the detection device 8 via the lens 6, the mirror 5, and the filter 7. The mirror 5 and the lens 6 are fixed to the moving plate 9. The moving plate 9 moves at a constant speed in the horizontal direction as the cam 10 and the transmission member 11 rotate, and the laser beam 4 is arranged inside the groove 2a as the moving plate 9, the mirror 5 and the lens 6 move horizontally. The fine particles 1 are sequentially scanned.

Such a DNA detection method and detection apparatus are described in Patent Document 1 (Japanese Patent Laid-Open No. 2000-346842).
Japanese Unexamined Patent Publication No. 2000-346842 (FIGS. 1 and 2)

  The diameter of the fine particles 1 is about 100 μm, but actually, the fine particles 1 have a tolerance. For this reason, the following problems arise.

  When the sample DNA is detected by the fluorescence reaction described above, it is necessary to know what probe the sample DNA is bound to. The probe is fixed to the fine particle 1, and the fine particle 1 is labeled with a fluorescent dye (having a fluorescent wavelength different from that of the fluorescent dye for labeling the sample DNA), thereby recognizing which probe is fixed to which fine particle. can do. Accordingly, if the detected sample DNA is detected by which of the fine particles 1 arranged in the groove 2a, the probe DNA has a complementary sequence. .

  For this reason, when the laser beam 4 is scanning the fine particles 1 in order, it is necessary to always know which fine particles are being scanned.

  The moving plate 9 of the fluorescence detection apparatus shown in FIG. 6 is moving horizontally at a constant speed. When the moving distance is divided by the average diameter of the fine particles, it is possible to estimate the position of the fine particles currently scanned from the fine particles at the inspection start point. For example, when the horizontal moving speed of the moving plate 9 is v and the elapsed time after scanning the fine particles at the measurement start point is Δt, the moving distance L of the laser beam 4 after scanning the fine particles at the measurement start point is L = vΔt. It is. As shown in FIG. 7A, when the diameter of the fine particles is uniform and the average diameter AD is equal to the diameter of each fine particle, the laser beam 4 estimated by dividing the moving distance L by the average diameter AD is scanned. The actual position coincides with the position of the fine particle T (the number of the fine particle from the measurement starting point S).

  However, since there is a tolerance in the diameter of the fine particles, the estimated value may be different from the relative position of the actually scanned fine particles. When the diameters of the fine particles vary, for example, in FIG. 7B, the estimated position of the fine particles T during scanning calculated by dividing the moving distance L of the laser beam 4 by the average diameter of the fine particles is 6 from the fine particles S at the measurement start point. In FIG. 7B, since there are many fine particles having a diameter smaller than the average diameter AD between the fine particles S and T, the seventh fine particle is actually scanned. Conversely, when there are many fine particles having a diameter larger than the average diameter AD between the fine particles S and the fine particles T, the estimated position of the fine particles T becomes larger than the actual position.

  Conventionally, there is no index of fine particles at the measurement start point, and it is difficult to accurately grasp the measurement start point.

  When the estimated value of the position of the microparticle currently scanned by the laser beam is shifted from the actual position and the position of the microparticle scanned by the laser beam cannot be grasped, a probe in which the sample DNA is fixed to the microparticle by fluorescence Even if the binding is detected, it is not possible to know which probe the detected sample DNA is bound to, and the base sequence of the sample DNA cannot be specified.

  An object of the present invention is to solve the above-described conventional problems, and to provide an inspection fine particle inspection method and an inspection fine particle inspection apparatus capable of accurately calculating the position of the inspection fine particle from a reference point. It is said.

The inspection fine particle inspection method of the present invention includes a flow path, an inflow port positioned upstream of the flow path, and an outflow port positioned downstream of the flow path. And arranging reference point fine particles, and arranging a plurality of the inspection fine particles on the downstream side of the reference point fine particles,
After the reference point fine particles are identified and an inspection start point is set, the state of the inspection fine particles is examined by an inspection means.

  In the present invention, the fine particles arranged in the flow path have two kinds of fine particles for inspection, which are the reference point fine particles serving as an index of the measurement start point, and inspection. By using the reference point fine particle as an index of the inspection start point, it is possible to accurately specify the relative position of the inspection fine particle arranged on the downstream side of the reference point fine particle from the reference point fine particle.

  In the present invention, it is preferable that a plurality of fine particle groups in which a plurality of the inspection fine particles are arranged on the downstream side of the reference point fine particles are arranged in the flow path from upstream to downstream.

  The deviation between the estimated position of the inspection fine particles and the actual position caused by the variation in the diameter of the fine particles is likely to occur as the relative distance from the reference point fine particles increases. According to the present invention, by inserting a plurality of the reference point fine particles between the arranged inspection fine particles, the deviation between the estimated position of the inspection fine particles and the actual position can be suppressed. Further, when different types of inspection fine particles are arranged in one flow path, the different inspection fine particles can be classified according to the reference point fine particles.

  In the present invention, the inspection means for examining the state of the fine particles is a spectroscopic means for detecting and analyzing, for example, fluorescence, phosphorescence, infrared rays, ultraviolet rays, and visible rays.

  In order to detect the inspection fine particles and the reference point fine particles, it is preferable to apply or contain a luminescent dye in one or both of the inspection fine particles and the reference point fine particles.

  At this time, the inspection fine particles and the reference point fine particles can be distinguished by making the emission wavelengths of the luminescent dyes applied or contained in the inspection fine particles different from the emission wavelengths of the light emitting dyes applied or contained in the reference point fine particles. .

  Furthermore, the inspection fine particles can be identified for each of the fine particle groups by applying or containing a light emitting dye having a different emission wavelength for each of the fine particle groups.

In the present invention, it is preferable to use a fluorescent dye as the luminescent dye.
It is preferable that a probe for capturing a specific specimen is bound to the test fine particles. A probe that captures a specific specimen is, for example, a DNA fragment having a complementary sequence when the specimen is DNA or RNA, and an antibody that specifically adsorbs when the specimen is a protein. Alternatively, using the principle of chromatography, it is also possible to configure inspection fine particles for detecting ionic molecules and sugar chains.

  In the present invention, after identifying the reference point fine particles and setting an inspection start point, the inspection means or the inspection base is moved in a direction parallel to the flow path, and the inspection means is relative to the inspection start point. By calculating the movement distance, it is possible to accurately measure the relative movement distance from the inspection start point.

  Further, it is preferable to calculate the number of inspection fine particles existing from the inspection start point to the current position of the inspection means by dividing the relative movement distance by the average diameter of the inspection fine particles.

  The relative movement distance is a relative distance from the reference point fine particle to the inspection fine particle being scanned. In the present invention, since the position of the inspection fine particle is estimated based on the relative distance from the reference point fine particle to the inspection fine particle being scanned, the accumulation of tolerances is suppressed, and the inspection during the scanning from the inspection start point is performed. It is possible to accurately estimate the position of the fine particles.

The inspection apparatus of the present invention includes a flow path, an inspection base having an inflow port located on the upstream side of the flow path, an outflow port located on the downstream side of the flow path, and an inside of the flow path. A plurality of fine particles arranged, the fine particles are composed of inspection fine particles and reference point fine particles, and a plurality of the inspection fine particles are arranged downstream of the reference point fine particles;
Furthermore, it has control means for identifying the reference point fine particles and setting an inspection start point, and inspection means for examining the state of the fine particles.

  In the flow path, it is preferable that a plurality of fine particle groups in which a plurality of the inspection fine particles are arranged on the downstream side of the reference point fine particles are arranged from upstream to downstream.

  Preferably, the inspection means detects the state of the fine particles by spectroscopic means for detecting and analyzing fluorescence, phosphorescence, infrared light, ultraviolet light, visible light, and the like.

It is preferable that a luminescent dye is applied or contained in the fine particles.
In particular, it is preferable that the luminescent dye coated or contained in the inspection fine particles and the luminescent dye coated or contained in the reference point fine particles have different emission wavelengths.

  Furthermore, it is preferable that the inspection fine particles are coated with or contain a luminescent dye having a different emission wavelength for each of the fine particle groups.

The luminescent dye is, for example, a fluorescent dye.
It is preferable that a probe for capturing a specific specimen is bound to the fine particle.

  It is preferable to have a moving means for moving the inspection means or the inspection substrate in a direction parallel to the flow path.

  At this time, after the inspection start point is set, the calculation means for calculating the relative movement distance of the inspection means in the flow path direction on the inspection substrate is connected to the control means, The relative distance from the reference point fine particles to the inspection fine particles being scanned can be calculated.

  In addition, the calculation means can calculate the number of inspection fine particles existing from the inspection start point to the current position of the inspection means by dividing the relative movement distance by the average diameter of the inspection fine particles.

  In the present invention, the fine particles arranged in the flow path have two kinds of fine particles for inspection, which are the reference point fine particles serving as an index of the measurement start point, and inspection. By using the reference point fine particle as an index of the inspection start point, it is possible to accurately specify the relative position of the inspection fine particle arranged on the downstream side of the reference point fine particle from the reference point fine particle.

  Further, a plurality of fine particle groups in which a plurality of the inspection fine particles are arranged on the downstream side of the reference point fine particles in the channel are arranged from upstream to downstream, that is, between the arranged inspection fine particles. By inserting a plurality of the reference point fine particles, it is possible to suppress the deviation between the estimated position of the inspection fine particles and the actual position due to the variation in the diameter of the fine particles. Further, when different types of inspection fine particles are arranged in one flow path, the different inspection fine particles can be classified according to the reference point fine particles.

  Further, in the present invention, since the position of the inspection fine particle is estimated based on the relative distance from the reference point fine particle to the inspection fine particle being scanned, accumulation of tolerances is suppressed, and It is possible to accurately estimate the position of the inspection fine particles.

  FIG. 1 is a partial perspective view of the appearance of an inspection plate substrate (inspection substrate), FIG. 2 is a plan view of the inspection plate shown in FIG. 1, and FIG. 3 is a conceptual diagram showing an embodiment of the inspection apparatus of the present invention. FIG. 4 and FIG. 4 are conceptual diagrams for explaining a method for detecting the relative position of the inspection fine particles from the reference point fine particles (inspection start point) using the inspection apparatus and inspection method of the present invention.

  The test plate P shown in FIG. 1 is for collecting blood or urine from a human body, for example, and performing a predetermined test by reacting these samples (specimens) with a predetermined reagent or the like. When the test plate is used as, for example, a DNA chip, the collected blood is used after being subjected to a predetermined treatment.

  The inspection plate P has a substantially rectangular shape having a predetermined thickness extending in the length direction (Y1-Y2 direction in the drawing) at right angles from both ends in the width direction (X1-X2 direction in the drawing), but other than the substantially rectangular shape. It may be in shape.

  The inspection plate P includes a plate substrate (inspection base) 12 and a lid body 13. The plate substrate 12 and the lid 13 are made of glass or resin. The plate substrate 12 and the lid 13 are made of a material having a predetermined fluorescence intensity. In particular, when the test plate P is used as a DNA chip, a protein chip, or the like, the test plate P is preferably made of a material that is almost transparent and has low fluorescence and excellent chemical resistance, such as quartz glass, It is formed of polydimethylsiloxane (PDMS), polymethyl methacrylate (PMMA) or the like.

  When the inspection plate P is formed of a resin, it is preferable to form the inspection plate P by injection molding. In some cases, the inspection plate P is formed on the upper surface 12a of the plate substrate 12 of the inspection plate P by hot pressing. The groove is formed with a high aspect ratio. When the inspection plate P is made of glass, it is formed by hot pressing.

  A single flow path 14 is formed in a concave shape on the upper surface 12a of the plate substrate 12 shown in FIGS. The Y1 side of the flow path 14 is located on the upstream side where the liquid such as the specimen flows in the flow path 14, and the Y2 side is located on the downstream side where the liquid such as the specimen flows in the flow path 14.

  As shown in FIGS. 1 and 2, an inflow port 15 connected to the flow path 14 is formed in a concave shape on the upstream side (Y1 side in the drawing) of the flow path 14. As shown in FIG. 1, the lid 13 is provided with an opening 13 a penetrating from the upper surface to the lower surface of the lid 13 at a position facing the inflow port 15. A substance can be put into the inlet 15.

  As shown in FIGS. 1 and 2, an outlet 16 connected to the flow path 14 is formed in a concave shape on the downstream side (Y2 side in the drawing) of the flow path 14. As shown in FIG. 1, the lid body 13 is provided with an opening 13 b penetrating from the upper surface to the lower surface of the lid body 13 at a position facing the outflow port 16. Such a substance can be discharged from the opening 13b.

  As shown in FIGS. 1 and 2, the flow path 14 is formed in a rectangular shape having a predetermined width dimension in the illustrated X1-X2 direction and extending in the illustrated Y1-Y2 direction. On the other hand, the planar shape of the inlet 15 and the outlet 16 is formed in a substantially circular shape, and the maximum width dimension of the inlet 15 and the outlet 16 is formed larger than the width dimension of the flow path 14. However, the shapes of the flow path 14, the inlet 15 and the outlet 16 are not limited to those shown in FIGS. 1 and 2, and may be other shapes.

  Basically, the flow path 14 has a function of allowing a single substance or a plurality of substances to flow from the upstream side (Y1 side in the figure) to the downstream side (Y2 side in the figure) or to mix a plurality of substances while flowing. On the other hand, the inlet 15 has a function of injecting one or a plurality of substances and feeding them into the flow path 14 or mixing a plurality of substances, and the outlet 16 discharges one or a plurality of substances. Or has a function of mixing a plurality of substances, and the way in which the flow path 14, the inflow port 15 and the outflow port 16 are used differs depending on the use mode.

  In the test plate P shown in FIG. 1, the flow path 14 is a measurement region for flowing a sample and measuring whether or not the sample and the probe have reacted during the flow. This is a region where the probe is injected, and the outlet 16 functions as a region where the specimen is discharged.

  A large number of fine particles 19 are inserted from the opening 13a shown in FIG. The fine particles 19 are spherical particles made of glass or resin.

  The fine particles 19 are composed of inspection fine particles 19a having probes attached to the surface and reference point fine particles 19b for setting an inspection start point. In the present embodiment, a group of fine particles G1, G2, G3, G4, G5, G6, G7, G8, and G9 in which a plurality of fine particles for inspection 19a are arranged on the downstream side of the reference point fine particles 19b are flowed. It arrange | positions from the upstream in the path | route 14 to the downstream.

  A probe that captures a specific specimen is, for example, a DNA fragment having a complementary sequence when the specimen is DNA or RNA, and an antibody that specifically adsorbs when the specimen is a protein. Alternatively, using the principle of chromatography, it is also possible to construct inspection fine particles for detecting ionic molecules and sugar chains.

  It is preferable that the emission wavelength of the fluorescent dye applied or contained in the inspection fine particles 19a and the fluorescent dye applied or contained in the reference point fine particles 19b are different.

  Further, each inspection fine particle 19a and each reference point fine particle 19b belong to any one of the fine particle groups G1, G2, G3, G4, G5, G6, G7, G8, and G9. It is preferable that the inspection fine particles 19a and the reference point fine particles 19b are coated with or contain a fluorescent dye having a specific emission wavelength for each group of fine particles to which they belong. Thereby, it is possible to identify which fine particle group the fine particles for inspection 19a and the reference fine particle 19b belong to among the fine particle groups G1, G2, G3, G4, G5, G6, G7, G8, and G9.

  A sample such as DNA collected from a person is flowed from the opening 13a shown in FIG. If DNA having a base sequence complementary to the probe is present in the sample, it is captured by a hybridization reaction with the probe.

  For example, the probe can be distinguished by a fluorescent dye having a different emission wavelength from the fluorescent dye added to the specimen, and the DNA contained in the specimen is detected by detecting to which probe the specimen DNA has undergone a hybridization reaction. Can be specified.

  The fluorescence intensity can be measured with a small CCD camera (detection means) 28 shown in FIG. 3, and it can be diagnosed whether the viewer has developed a specific disease (for example, cancer) from the fluorescence intensity. .

  Here, each dimension will be described. The width dimension T1 and the height dimension H of the flow path 14 are formed to be about 50 μm to 100 μm. The average diameter of the inspection fine particles 19a and the reference point fine particles 19b is 50 μm to 100 μm, and the tolerance is ± 5 to 10%.

  FIG. 3 is a conceptual diagram showing an embodiment of the detection apparatus of the present invention. The detection device according to the present embodiment irradiates the fine particles 19 arranged in the flow path 14 of the plate substrate 12 with laser light.

  A laser beam 24 emitted from a laser light source 23 is irradiated onto the fine particles 19 on the plate substrate 12 via a mirror 25 and a lens 26, and is bound to a sample hybridized with a fluorescent dye or a probe in the fine particles 19. Excites the fluorescent dye. The plate substrate 12 is the one shown in FIGS. 1 and 2, and the fine particles 19 are the fine particles G1, G2, G3, G4, G5, G6, G7, G8, and G9 as shown in FIG. It is arranged from upstream to downstream in the passage 14. In the fine particle groups G1, G2, G3, G4, G5, G6, G7, G8, and G9, a plurality of the inspection fine particles 19a are arranged on the downstream side of the reference point fine particles 19b.

  The excited fluorescent dye emits fluorescent light R having a wavelength specific to the fluorescent dye, and this fluorescent light R is detected by the CCD camera 28 through the lens 26, the mirror 25, and the filter 27. The mirror 25 and the lens 26 are fixed to a moving plate (moving means) 29. The moving plate 29 moves at a constant speed in the horizontal direction as the cam 30 and the transmission member 31 rotate, and the laser beam 24 is arranged inside the flow path 14 as the moving plate 29, the mirror 25, and the lens 26 move horizontally. The formed fine particles 19 are sequentially scanned. In this embodiment, the mirror 25 and the lens 26 of the inspection means are moved in the direction parallel to the flow path 14 by the moving plate 29, but the plate substrate 12 may be moved in the direction parallel to the flow path 14. Good.

  In the inspection apparatus of the present invention, the control means 32 for identifying the reference point fine particles 19b from the fine particles 19 and setting the inspection start point is connected to the CCD camera 28.

  The control means 32 detects the difference in fluorescence wavelength between the reference point fine particles 19b and the inspection fine particles 19a and distinguishes the reference point fine particles 19b from the inspection fine particles 19a. In addition, the calculation means 33 is connected to the control means 32. The calculation means 32 recognizes the reference point fine particles 19b and sets the inspection start point, and then the relative movement distance in which the mirror 25, the lens 26, and the laser light 24 move on the plate substrate 12 in the flow path direction (Y2 direction in the drawing). Is calculated.

  In the present invention, by using the reference point fine particle 19b as an index of the inspection start point, the position of the inspection fine particle 19a arranged on the downstream side of the reference point fine particle 19b from the reference point fine particle 19b can be accurately specified. become.

  When the sample DNA captured by the probe is detected by a fluorescent reaction, it is necessary to know what probe the sample DNA is bound to. For example, the inspection fine particles 19a can be identified by labeling with a fluorescent dye (having a fluorescence wavelength different from that of the fluorescent dye for labeling the sample DNA), and the type of probe fixed to the inspection fine particles 19a can be recognized. it can. Therefore, if the detected sample DNA is detected by which of the test fine particles 19a among the test fine particles 19a arranged in the flow path 14, the sample DNA has a complementary sequence to which probe. You can see that

  For this reason, when the laser beam 24 scans the fine particles 19 in order, it is necessary to always know which fine particles 19 are being scanned.

  The moving plate 29 of the fluorescence detection apparatus shown in FIG. 3 is moving horizontally at a constant speed. As shown in FIG. 4, for example, when the horizontal movement speed of the moving plate 29 is v and the elapsed time after scanning the fine particles at the measurement start point is Δt, the laser light 24 after scanning the fine particles at the measurement start point The relative movement distance L is L = vΔt. In the present invention, after the reference point fine particle 19b is recognized and the inspection start point is set, the relative movement distance L from the inspection start point (the position of the reference point fine particle 19b) is divided by the average diameter AD of the fine particle 19, It is estimated what position the inspection fine particle 19a currently scanned is positioned from the inspection start fine particle (reference point fine particle 19b).

  The deviation between the estimated position and the actual position of the inspection fine particles 19a caused by the variation in the diameter of the fine particles 19 tends to occur as the distance from the reference point fine particles 19b increases. In the present embodiment, the relative movement distance of the laser beam 24 is reset when the reference point fine particle 19b is detected, and the relative movement distance L of the laser beam 24 is measured using the position of the reference point fine particle 19b as the inspection start point. As in the present embodiment, by inserting a plurality of reference point fine particles 19b between the inspection fine particles 19a, it is possible to reset and measure again before the relative movement distance L of the laser light 24 increases. In addition, it is possible to suppress the difference between the estimated position and the actual position of the inspection fine particles 19a from the reference point fine particles 19b by suppressing the accumulation of tolerances. Further, the reference point fine particles 19b can be classified according to the types of probes to which the inspection fine particles 19a are coupled.

  In the present invention, since it is possible to accurately grasp the position of the fine particle 19 currently scanned by the laser beam, it is detected by fluorescence that the sample DNA is bound to the probe fixed to the test fine particle 19a. It is possible to accurately grasp which probe the detected sample DNA has bound to. Therefore, the base sequence of the sample DNA can be accurately specified.

  In this embodiment, in order to identify the reference point fine particles 19b and the inspection fine particles 19a, the reference point fine particles 19b and the inspection fine particles 19a are fluorescently labeled, and the fine particles are identified by the difference in the respective fluorescence wavelengths. However, the reference point fine particles 19b and the inspection fine particles 19a may be identified using a label such as phosphorescent light, infrared light, ultraviolet light, or visible light.

Further, the reference point fine particles 19b and the inspection fine particles 19a can be identified not by the wavelengths of fluorescence, phosphorescence, infrared rays, ultraviolet rays, and visible rays but by the intensity of the rays. In this case, for example, fluorescent dyes are blended in different proportions in the inspection fine particles 19a and the reference point fine particles 19b.
The sample DNA can also be labeled with phosphorescence or ultraviolet light.

A perspective view of an inspection plate used in the present invention, A plan view of an inspection plate in which fine particles used in the present invention are arranged, The conceptual diagram which shows embodiment of the test | inspection apparatus of this invention, Schematic diagram showing the state of fine particles under inspection, A plan view of an inspection plate in which fine particles of the conventional invention are arranged, Conceptual diagram of a conventional inspection device, (A) is a schematic diagram showing the state of an ideal fine particle under inspection with no tolerance in diameter, (b) is a schematic diagram showing the state of an actual fine particle under inspection having a tolerance in diameter,

Explanation of symbols

12 Plate substrate 14 Flow path 15 Inlet 16 Outlet 19 Fine particle 19a Fine particle 19b for inspection Reference point fine particle 23 Laser light source 24 Laser light 28 CCD camera 29 Moving plate 32 Control means 33 Calculation means

Claims (21)

A reference point fine particle is disposed inside the flow path of the inspection substrate having a flow path, an inlet located upstream of the flow path, and an outlet located downstream of the flow path, and the reference A plurality of the inspection fine particles are arranged downstream of the point fine particles,
An inspection method in which after the reference point fine particles are identified and an inspection start point is set, the state of the inspection fine particles is checked by an inspection unit.   2. The inspection method according to claim 1, wherein a plurality of fine particle groups in which a plurality of the inspection fine particles are arranged on the downstream side of the reference point fine particles are arranged in the flow channel from upstream to downstream.   3. The inspection method according to claim 1, wherein the inspection means for examining the state of the fine particles is a spectroscopic means.   The inspection method according to claim 3, wherein a luminescent dye is applied or contained in one or both of the inspection fine particles and the reference point fine particles.   The inspection method according to claim 4, wherein the emission wavelength of the luminescent dye applied or contained in the inspection fine particles and the luminescent dye applied or contained in the reference point fine particles are made different.   The inspection method according to claim 4 or 5, wherein the luminescent dye having a different emission wavelength is applied or contained in the inspection fine particles for each of the fine particle groups.   The inspection method according to claim 4, wherein a fluorescent dye is used as the luminescent dye.   The inspection method according to claim 1, wherein a probe that captures a specific specimen is bound to the inspection microparticles.   After identifying the reference point fine particles and setting the inspection start point, the inspection means or the inspection substrate is moved in a direction parallel to the flow path, and the relative movement distance of the inspection means from the inspection start point is calculated. The inspection method according to any one of claims 1 to 8.   The inspection method according to claim 9, wherein the number of inspection fine particles existing from the inspection start point to the current position of the inspection means is calculated by dividing the relative movement distance by the average diameter of the inspection fine particles. A test substrate having a flow path, an inlet positioned upstream of the flow path, and an outlet positioned downstream of the flow path, and a plurality of fine particles arranged inside the flow path. The fine particles are composed of inspection fine particles and reference point fine particles, and a plurality of the inspection fine particles are arranged downstream of the reference point fine particles,
The inspection apparatus further comprises control means for identifying the reference point fine particles and setting an inspection start point, and inspection means for examining the state of the fine particles.   The inspection apparatus according to claim 11, wherein a plurality of fine particle groups in which a plurality of the inspection fine particles are arranged on the downstream side of the reference point fine particles are arranged in the flow path from upstream to downstream.   The inspection apparatus according to claim 11 or 12, wherein the inspection means detects the state of the fine particles by spectroscopic means.   The inspection apparatus according to claim 13, wherein a luminescent dye is applied or contained in the fine particles.   The inspection apparatus according to claim 14, wherein the luminescent dye applied or contained in the inspection fine particles and the luminescent dye applied or contained in the reference point fine particles have different emission wavelengths.   The inspection device according to claim 14 or 15, wherein the inspection fine particles are coated with or contain the luminescent dye having a different emission wavelength for each of the fine particle groups.   The inspection apparatus according to claim 14, wherein the luminescent dye is a fluorescent dye.   The inspection apparatus according to claim 11, wherein a probe for capturing a specific specimen is bound to the fine particles.   The inspection apparatus according to claim 11, further comprising a moving unit that moves the inspection unit or the inspection substrate in a direction parallel to the flow path.   An arithmetic means for calculating a relative movement distance that the inspection means moves in the direction of the flow path on the inspection substrate after the inspection start point is set is connected to the control means. The inspection apparatus in any one of.   21. The calculation means calculates the number of inspection fine particles existing from the inspection start point to the current position of the inspection means by dividing the relative movement distance by the average diameter of the inspection fine particles. Inspection equipment.

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