JP6184292B2 - Analyzer using compact disc type microchip - Google Patents

Description

  The present invention relates to an analyzer using a compact disk type microchip.

  Many chemical substances created by mankind are harmful to the human body. In recent years, environmental pollution caused by harmful chemical substances such as dioxins has become a problem. For example, when looking at the eating habits, problems due to residual agrochemicals, bacteria such as O-157, or allergens such as buckwheat, which threaten food safety, are becoming serious. The epidemic of emerging and re-emerging infectious diseases such as new influenza and tuberculosis has become a social problem. In order to solve such problems, analysis of chemical substances, bacteria or viruses is indispensable. Furthermore, there is an increasing demand for analyzing stress markers in the human body in order to reduce health problems associated with high stress societies. However, conventional analyzers used in environmental measurements, food inspections, medical inspections, etc. are extremely expensive, take a lot of time for measurement, and are portable that can be easily measured on-site or bedside. It is not a simple analyzer. Under such circumstances, there is a need for the development of a new small analysis system that can be measured easily, quickly and with high sensitivity by anyone in the field.

  In recent years, a microchemical analysis system (μTAS) has attracted attention as an analysis system that can meet such demands. μTAS forms a fine channel (microchannel) on a palm-sized glass or plastic substrate (microchip), and performs a series of chemical analysis operations such as mixing, reaction, separation, and detection in the microchannel. System (see, for example, Patent Document 1). μTAS not only reduces the size of the analyzer, but also has many advantages such as reducing the amount of reagents and samples, reducing measurement time, and reducing costs, and is expected to develop in the future as an innovative new technology. ing. So far, research on integration of separation fields and reaction fields has been actively conducted, and enzyme sensors, electrophoresis, liquid chromatography, immunoassay methods using microchips, etc. have been developed. However, progress has been made in the integration of detection systems and liquid delivery systems, and in any μTAS, peripheral devices such as pumps and valves used for feeding sample solutions and reagent solutions, and lasers and microscopes used for detection are large. Have the common problem of being expensive and expensive.

  The present inventors have so far developed a surface plasmon resonance (SPR) sensor using a compact disk (CD) type microchip. This is because a large number of microchannels and reservoirs are formed on one CD, and the sample solution in the reservoir is introduced into the microchannel using the centrifugal force generated by the rotation of the CD, and the analyte substance in the sample solution is introduced. After separation by interaction with a receptor protein immobilized on the inner wall of the microchannel, detection is performed by a reflection-type SPR sensor of the grating type. This sensor uses the centrifugal force generated by the rotation of the CD-type microchip to send the sample solution, which eliminates the need for a pump or valve for feeding the liquid, and is useful for downsizing the liquid feeding system. . In addition, since detection is performed using the SPR phenomenon, there is an advantage that labeling of a sample with a dye or an enzyme is unnecessary.

JP 2004-156925 A

  However, the SPR sensor using the above-described CD type microchip still has problems to be improved. That is, in terms of sensitivity, the fluorescence method and the thermal lens method are still superior. The present inventors have developed a small and inexpensive fluorescence analysis system combining a CD-type microchip and an LED-induced fluorescence detection method, and succeeded in measuring immunoglobulin A (IgA) by enzyme immunoassay (ELISA). . However, in this system, since a commercially available LED is used as a light source, there is a further problem of further downsizing the apparatus and improving measurement reproducibility.

  The present invention has been made to solve the above-described problems, and an object of the present invention is to provide a smaller and more highly reproducible analyzer.

  One form of the present invention for achieving the above object is an analyzer used for analyzing a sample and capable of feeding the solution radially outward from the center using centrifugal force when rotating. One or two or more storage parts for storing the liquid, an analysis part that is arranged radially outside the storage part and serves as an analysis field for the sample sent from the storage part, and a liquid feed that connects the storage part and the analysis part A compact disc-type microchip including at least a flow path for use, a light-emitting device including organic electroluminescence as a light source that emits light toward the analysis unit, and a light-receiving device including a light-receiving unit that receives light from the analysis unit, This is an analyzer using a compact disk type microchip.

  Another embodiment of the present invention is an analyzer using a compact disk type microchip further comprising a first rotation mechanism for rotationally driving the compact disk type microchip.

  Another embodiment of the present invention uses a compact disc type microchip that further includes a second rotation mechanism that rotationally drives the organic electroluminescence, and that allows the organic electroluminescence to rotate in synchronization with the compact disc type microchip. It is an analysis device.

  Another embodiment of the present invention is an analyzer using a compact disk type microchip that further includes a third rotation mechanism that rotationally drives the light receiving part, and that allows the light receiving part to rotate in synchronization with the compact disk type microchip. .

  Another embodiment of the present invention is an analyzer using a compact disk type microchip having a drive unit common to the first rotation mechanism, the second rotation mechanism, and the third rotation mechanism.

  Another embodiment of the present invention is an analyzer using a compact disk type microchip in which organic electroluminescence is disposed below the compact disk type microchip.

  Another embodiment of the present invention is an analyzer using a compact disk type microchip in which organic electroluminescence is disposed above the compact disk type microchip.

  Another embodiment of the present invention is an analyzer using a compact disc type microchip for use in a detection unit to detect a substance based on specific binding between an antigen and an antibody in an analysis unit.

  Another embodiment of the present invention is an analyzer using a compact disk type microchip in which at least an antibody capable of binding to an antigen in a sample is disposed in an analysis unit.

  According to the present invention, it is possible to provide a smaller analyzer with high measurement reproducibility.

FIG. 1 shows a schematic diagram of an analyzer using a compact disk type microchip according to an embodiment of the present invention. FIG. 2 shows a plan view and a side view of the CD type microchip of FIG. 3 is a plan view (3A) of one block in the CD-type microchip of FIG. 2, a sectional view taken along line AA (3B) in the plan view, and a sectional view taken along line BB in the plan view (3C). Are shown respectively. FIG. 4 shows an example of a manufacturing process of the CD type microchip of FIG. FIG. 5 shows a manufacturing process of the CD type microchip of FIG. FIG. 6 shows a manufacturing process following the manufacturing process of FIG. FIG. 7 is a view for explaining the liquid feeding principle using the CD type microchip of FIG. FIG. 8 shows a perspective view of the organic EL of FIG. FIG. 9 shows a typical flow of analysis using the analyzer of FIG. 1. One is a method (9A) in which the analysis is performed after rotating the CD-type microchip and stopping the rotation. Shows a method (9B) for performing analysis while rotating a CD-type microchip. FIG. 10 shows a modification of the analyzer of FIG. 1, in which the light emitting device and the light receiving device can be rotated in synchronization with the rotation of the CD type microchip, and the light emitting device is arranged below the CD type microchip. A schematic diagram (10A) and a simplified configuration diagram (10B) of an analyzer of the type to be used are shown. FIG. 11 is a modified example different from the analysis apparatus of FIG. 10, in which the light emitting device and the light receiving device can be rotated in synchronization with the rotation of the CD microchip, and the organic EL is disposed above the CD microchip. A schematic diagram (11A) and a simplified configuration diagram (11B) of an analyzer of the type disposed in FIG. FIG. 12 shows a condition (12A) for introducing a fluorescent reagent by changing the concentration and volume in five reservoirs formed in one block in the CD-type microchip in FIG. 3, and fluorescence detection under that condition. The result (12B) which performed the performance evaluation of the system is shown. FIG. 13 is a diagram for explaining a fluorescence detection method for immunoglobulin A, which is positioned as a stress marker contained in saliva, using the analyzer of FIG. 1. A state (13B) of the analysis unit that changes in accordance with the liquid feeding is shown. FIG. 14 shows the measurement method (14A) and measurement result (14B) of immunoglobulin A using the analyzer of FIG.

  Next, a preferred embodiment of an analyzer using a compact disk type microchip according to the present invention will be described with reference to the drawings.

<1. Outline of analyzer>
FIG. 1 shows a schematic diagram of an analyzer using a compact disk type microchip according to an embodiment of the present invention.

  An analyzer 1 (hereinafter simply referred to as an “analyzer”) 1 using a compact disk type microchip according to this embodiment is used for analyzing a sample and utilizes a centrifugal force when rotating from the center. It is a device that can feed liquid radially outward. The analyzer 1 includes a compact disc type microchip (hereinafter, referred to as “CD type microchip” as appropriate) 2 and organic electroluminescence (hereinafter, referred to as a light source) that emits light toward the analysis part of the CD type microchip 2. And a light receiving device 5 including a light receiving unit 16 that receives light from the analysis unit. The analysis apparatus 1 preferably further includes a first rotation mechanism 3 that rotationally drives the CD type microchip 2. In FIG. 1, the light receiving unit 16 is drawn so as to be opposed to the organic EL 13 with the analysis unit interposed therebetween. However, the light receiving unit 16 is not limited to this position, and is similar to the organic EL 13 and is a CD microchip. It may be arranged below 2. The light-emitting device 4, the light-receiving device 5, and the control device 6 described below are depicted as floating in the air in the figure, but are not actually floating in the air and may be arranged at any location. Interpreted to meaning.

  The light receiving device 5 is preferably connected to the control device 6 and can transmit information based on the received light to the computer under the control of the control device 6. The first rotating mechanism 3 includes a motor 10 as a driving unit and a rotating shaft 11 connected thereto. The rotating shaft 11 has a structure that can be connected to the approximate center of the CD-type microchip 2. For this reason, the CD type microchip 2 can be rotated by starting the motor 10 and rotating the rotating shaft 11 in a predetermined direction.

  The light emitting device 4 includes at least an organic EL 13 and is suitably provided with a power source 12 in this embodiment. In addition, the light emitting device 4 preferably includes a band pass filter 14 and a hemispherical lens 15 for sequentially filtering light from the organic EL 13 on the CD-type microchip 2 side from the organic EL 13. As the power source 12, a constant current power source can be suitably used. The band pass filter 14 is a filter for transmitting only a wavelength in a specific range, and for example, an excitation filter (560AF55) manufactured by Omega Optical can be used. When the band pass filter 14 is used, it is possible to effectively prevent the background signal from increasing due to the broad half-value width of the emission spectrum of the organic EL 13 and overlapping with the fluorescence of the sample. The hemispherical lens 15 is a lens for condensing light from the organic EL 13. Details of the organic EL 13 will be described later.

  The light receiving device 5 includes at least a light receiving unit 16, and in this embodiment, a band pass filter 17 is preferably provided on the CD microchip 2 side from the light receiving unit 16. As the light receiving unit 16, a CCD camera, a photomultiplier tube (PMT), a photodiode (PD), an organic photodiode (OPD), and the like can be exemplified. In particular, a CCD camera can be preferably exemplified. Hereinafter, an example using a CCD camera as the light receiving unit 16 will be described unless otherwise specified. The CCD camera enables recording and analysis of signals based on light reception by means of CCD camera software stored in the memory on the control device 6 side. For example, a CCD camera (TCN-1304-U) manufactured by Mightex is used. Can do. The bandpass filter 17 is a filter for transmitting only a wavelength in a specific range of the fluorescence spectrum from the sample. For example, a fluorescent filter (645AF75) manufactured by Omega Optical can be used.

  When the analysis apparatus 1 having the above configuration is used, the sample collected in the analysis unit of the CD-type microchip 2 can be illuminated from the organic EL 13 and the sample can be analyzed from information obtained by light reception from the sample. Here, what is important is that a suitable example of the surface emitting means called the organic EL 13 is used. Since light with excellent uniformity of in-plane light intensity can be applied to the sample in the analysis unit from the organic EL 13, analysis with high reproducibility is possible even if the illumination position in the analysis unit is not strictly set. . In addition, since the analysis apparatus 1 includes the first rotation mechanism 3, the analysis can be performed by emitting the organic EL 13 after the CD type microchip 2 is rotated and the sample is moved to the analysis unit. First, the organic EL 13 can emit light while rotating the CD-type microchip 2, and the change over time in the analysis value due to the movement of the sample can be examined.

<2. Structure of CD type microchip>
FIG. 2 shows a plan view and a side view of the CD type microchip of FIG.

  The CD-type microchip 2 used in this embodiment is used for analyzing a sample and has a structure capable of feeding liquid from the center to the outside in the radial direction using a centrifugal force when the sample is rotated. The CD-type microchip 2 is a disk-shaped microchip having a hole 25 in the center, and is divided into eight substantially fan-shaped blocks 30 in plan view. Each block 30 has a channel structure with the same shape. For this reason, the analysis up to N = 8 can be simultaneously performed using the CD type microchip 2. However, the number of blocks 30 is not limited to eight, and the number of blocks 30 is not limited as long as it is at least one. The CD type microchip 2 is preferably formed by laminating a first disk 34 and a second disk 35 made of a light-transmitting material. In this embodiment, the first disk 34 is made of polydimethylsiloxane (a kind of silicone resin). However, instead of the above-described silicone resin, another light-transmitting material can be selected as the material constituting the first disk 34. As the material, for example, acrylic resins, polycarbonate resins, cycloolefin resins, polystyrene resins, polyester resins, urethane resins, vinyl chloride resins and other synthetic resins, or glass can be used.

  In this embodiment, the second disk 35 is made of polycarbonate resin. However, as the material constituting the second disk 35, another light-transmitting material can be selected instead of the polycarbonate resin. As the material, for example, synthetic resin such as acrylic resin, silicone resin, polyester resin, urethane resin, vinyl chloride resin, or glass can be used. In the present application, “translucent” is broadly interpreted to mean that light can be transmitted regardless of whether it is colored or colorless and the light transmittance is large.

  3 is a plan view (3A) of one block in the CD-type microchip of FIG. 2, a sectional view taken along line AA (3B) in the plan view, and a sectional view taken along line BB in the plan view (3C). Are shown respectively.

  The CD-type microchip 2 includes at least one or more storage units 41, 42, 43, 44, 45 (hereinafter referred to as “storage unit 41”) for storing a sample and a radial direction from the storage unit 41 or the like. There are provided at least an analysis unit 46 which is disposed outside and serves as an analysis field for a sample sent from the storage unit 41 and the like, and a liquid-feeding flow channel 48 which connects the storage unit 41 and the analysis unit 46. More preferably, in this embodiment, as shown in (3A), each block 30 of the CD-type microchip 2 includes five storage parts 41 and the like, one analysis part 46, and one A waste liquid part 47 and a plurality of liquid-feeding channels 48 are provided. The flow path 48 connects the storage units 41 and the like, the storage units 41 and the like, the analysis unit 46, and the waste liquid unit 47, and the analysis unit 46 and the waste liquid unit 47, respectively. The storage unit 41 and the like are regions for storing at least a sample to be measured, and are a thin, substantially cylindrical space. In this embodiment, the storage portion 41 and the like have the same shape, but some or all of them may have different shapes. In this embodiment, the storage part 41 and the like are arranged in the order of the storage part 41, the storage part 42, the storage part 43, the storage part 44, and the storage part 45 from the radially outer side to the inner side of the CD-type microchip 2. ing. For this reason, when the CD microchip 2 is rotated, as the number of rotations is increased, liquid is first fed from the radially outermost storage section 41, then from the storage section 42, and then from the storage section 43. Then, liquid can be fed from the reservoir 44 and finally from the radially innermost reservoir 45.

  However, a part or all of the storage unit 41 and the like can be arranged at the same distance from the center of the CD-type microchip 2. As a result, the solutions that have been separated and stored can be simultaneously fed and mixed in the analysis unit 46. In addition to the sample to be measured, the storage unit 41 and the like can store other solutions such as a cleaning solution, a solution containing an antibody, and a solution containing various reagents. Furthermore, the number is not limited as long as there is at least one storage unit 41 and the like, instead of five. Hereinafter, “radial direction” means the radial direction of the CD-type microchip 2.

  The analysis unit 46 is a region arranged radially outside the storage unit 41 and the like, and is a region for illuminating a solution such as a sample sent from the storage unit 41 and the like. The reason why the analysis unit 46 is arranged on the outer side in the radial direction from the storage unit 41 and the like is to send the solution to the analysis unit 46 using the centrifugal force when the CD microchip 2 is rotated. The waste liquid part 47 is an area for collecting a liquid to be finally discarded from the analysis part 46 or the storage part 41 or the like. When the waste liquid part 47 is arranged radially outside the analysis part 46, the CD-type microchip 2 can be rotated after the measurement, and all the solutions can be collected in the waste liquid part 47. However, the waste liquid part 47 may be provided radially inward of the analysis part 46 or at the same radial distance as the analysis part 46. In that case, the measurer can manually collect the solution in the waste liquid portion 47 by tilting the CD microchip 2. Further, the waste liquid part 47 is not an essential area for the CD-type microchip 2 and may not be provided. In that case, the waste liquid is preferably discharged from the storage unit 41 or the analysis unit 46 after the measurement. In this sense, the flow channel 48 may be at least connected from the storage unit 41 or the like to the analysis unit 46. Each of the channels 48 on the radially inner side and the radially outer side of the storage unit 41 and the like is preferably provided with one enlarged diameter portion 49 in which each channel 48 is slightly thicker at a location very close to the storage unit 41 and the like. . The enlarged diameter portion 49 has a kind of valve function. By changing the diameter of the enlarged diameter portion 49, it is possible to change the number of rotations when the solution flows out from the storage portion 41 or the like. However, the enlarged diameter portion 49 is not an essential configuration and may not be provided.

  As shown in (3B), the flow path 48 is formed inside the CD microchip 2 in the thickness direction. The reservoir 45 is formed in the thickness direction of the CD-type microchip 2 and has an opening 50 on the upper surface thereof, like the channel 48. Although not explicitly shown in (3B), the other reservoirs 41, 42, 43, and 44 also have an opening 50 on the upper surface thereof. The opening 50 is preferably formed in order to supply a solution (including a sample) to the reservoir 41 and the like. The opening 50 is preferably covered with a film or the like (not shown) after supplying the solution to the reservoir 41 or the like. This is because it is necessary to prevent the solution from jumping out of the storage portion 41 or the like or dust or dust from the outside when the CD microchip 2 is rotated for measurement. In this embodiment, the storage section 41, the analysis section 46, the waste liquid section 47, and the flow path 48 are formed by recesses that are recessed inwardly from the facing surface of the second disk 35 in the first disk 34. The recesses on the second disk 35 side may be formed, or the recesses on the first disk 34 side and the second disk 35 side may be formed together. Further, the first disk 34 is provided with a concave portion that forms any one of the storage unit 41, the analysis unit 46, the waste liquid unit 47, and the flow path 48, and the second disk 35 is provided with a portion other than the part. You may provide the recessed part to form. That is, the CD-type microchip 2 includes a first disk 34 in which at least one of the storage unit 41 and the like, the analysis unit 46, the waste liquid unit 47, and the flow channel 48 is formed as a concave portion that is recessed inward. By stacking with one disk 34, the storage section 41 and the like, the analysis section 46, the waste liquid section 47, and the second disk 35 capable of forming a flow path 48 are stacked. However, in order to further simplify the process of forming the recesses, any one of the first disk 34 and the second disk 35 is provided with the respective recesses for the storage part 41, the analysis part 46, the waste liquid part 47, and the flow path 48. Is more preferable. When the concave portions for the storage portion 41, the analysis portion 46, the waste liquid portion 47, and the flow path 48 are formed on the first disc 34, there is no limitation on the thickness of the first disc 34 and the depth of the concave portion. For example, the thickness of the first disk 34 can be set in the range of 1 to 3 mm, and the depth of the recess can be set in the range of 20 to 80 μm. The thickness of the second disk 35 is not limited at all, but can be set within a range of 1 to 3 mm, for example.

  As shown in (3C), the waste liquid part 47 is formed inside the thickness direction of the CD-type microchip 2 and has an opening 51 on the upper surface thereof, like the storage part 45 described above. The opening 51 is preferably always covered with a film or the like (not shown) except during waste liquid. In this embodiment, the analysis unit 46 and the flow path 48 become a closed space after the first disk 34 and the second disk 35 are stacked, and do not have a dedicated opening. In addition, an opening may be provided in at least one of the flow channel 48. At the time of measurement, the entire upper surface of the CD-type microchip 2 or the upper surface of each block 30 is covered with a film or the like (not shown), and the storage unit 41 and the like, the analysis unit 46, the waste liquid unit 47, and the flow path 48. All may be provided with openings.

  Both the first disk 34 and the second disk 35 are translucent. Therefore, the light that has entered the analysis unit 46 from the first disk 34 side of the CD-type microchip 2 passes through the analysis unit 46 and the second disk 35. However, as long as at least the light entrance window and the light exit window of the analysis unit 46 of the first disk 34 are translucent, other portions can be light-shielding.

<3. Manufacturing method of CD type microchip>
FIG. 4 shows an example of a manufacturing process of the CD type microchip of FIG. 5 and 6 show a manufacturing process of the CD type microchip of FIG.

Step 101
First, a circular flat plate 60 necessary for manufacturing the first disk 34 in a plan view is prepared (5A). As the flat plate 60, for example, a silicon wafer can be preferably used. In this embodiment, a silicon wafer having a diameter of 6 inches is preferably used.

Step 102
Next, a negative photoresist is supplied on one surface of the flat plate 60, and a coating 61 having a uniform thickness is formed by means such as spin coating (5B). As the negative photoresist, for example, an epoxy resin-based cationic polymer, an oxetane resin-based cationic polymer, or the like can be used. In this embodiment, a negative photoresist having a property of promoting the crosslinking of the exposed portion is preferably used. On the contrary, a positive photoresist capable of dissolving and removing the exposed portion may be used. However, as a constituent material of the first disk 34 of the CD type microchip 2, a negative photoresist excellent in resistance to chemicals, mechanical strength and heat resistance, particularly SU-8 (manufactured by Kayaku Microchem) is used. It is more preferable to use it. After forming the coating film 61 on the flat plate 60, it is preferable to heat at around 100 ° C. to remove the organic solvent in the negative photoresist, and then to cool to room temperature.

Step 103
Next, a photomask 62 is placed on the negative photoresist film 61 (5C). The photomask 62 and the coating film 61 are preferably adhered and fixed using, for example, a vacuum clamp. The photomask 62 has a channel region of the CD microchip 2 as a light-transmitting region and a region other than the channel region as a light-shielding region. In the case where the coating film 61 is formed with a positive photoresist, the light transmitting region and the light shielding region of the photomask 62 are opposite to the previous configuration.

Step 104
Next, ultraviolet rays (UV) are irradiated from above the photomask 62 (5C). In this embodiment, a light box (LIGHT BOX BOX-W9B, manufactured by Sanhayato) is preferably used for irradiation for about 1 minute.

Step 105
Next, after curing the exposed region, the photomask 62 is removed from the flat plate 60 with the coating film 61, and the uncured resist in the non-irradiated region of ultraviolet rays is removed using a developer (5D). Prior to the removal of the uncured resist, the flat plate 60 with the coating film 61 may be heated at around 100 ° C. to enhance the fixation of the exposed region. As the developer, for example, a developer (manufactured by Kayaku Microchem, SU-8 Developer) can be preferably used. After removal of the uncured resist, the flat plate 60 with the coating film 61 may be washed with an organic solvent such as isopropanol (IPA), and further heated at around 200 ° C. to completely remove the organic solvent. Through such a process, a template in a form in which the channel region 64 for the CD type microchip 2 is protruded and the outside of the channel region 63 is recessed is completed.

Step 106
Next, the liquid polymer 65 is applied on the template (5E). As the liquid polymer 65, a material that becomes the first disk 34 after curing is used. As the material, for example, a polyalkylsiloxane, more preferably a material mainly containing polydimethylsiloxane can be used. More specifically, the main agent of polydimethylsiloxane (Toray Dow Corning, SILPO T184 W / C BASE) and its curing agent (Toray Dow Corning, SILPO T 184 W / C CURING AGENT) in a weight ratio of 10: The prepolymer mixed in 1 can be suitably used as the liquid polymer 65. When applying the liquid polymer 65 on the template, from the viewpoint of forming the first disk 34 with high accuracy, the template is placed in a mold having a cylindrical space on the bottom surface that is substantially the same as the template, and the liquid polymer 65 is applied from above. Is preferred.

Step 107
Next, the liquid polymer 65 is cured (5E). Prior to curing, it is preferable to degas the liquid polymer 65 under normal pressure or reduced pressure. In curing, if the liquid polymer 65 is a thermosetting resin, heating is preferably performed. For example, when polydimethylsiloxane is used, it is preferable to heat at 50 to 70 ° C. On the other hand, when the room temperature curable liquid polymer 65 is used, heating is not required.

Step 108
Next, the cured light-transmitting polymer 34 ′ having a flow path formed is removed from the template (5F).

Step 109
Next, the openings 50 and 51 are formed in the reservoir 41 and the waste liquid portion 47 of the translucent polymer 34 ′ in which the flow path has been formed, respectively, and the hole 25a is formed in the center in the plane (5G). Thus, the first disk 34 is completed.

Step 110
Next, a circular translucent flat plate 35 necessary for manufacturing the second disk 35 in a plan view is prepared. For example, a plate made of polycarbonate resin can be used as the flat plate 35.

Step 111
Next, a hole 25b is formed in the in-plane center of the flat plate 35 (6A).

Step 112
Next, the first disk 34 and the flat plate (second disk) 35 are bonded together (6B). When pasting, any means such as an adhesive, a double-sided tape, and fitting may be used. Thus, the CD type microchip 2 is completed.

  Each step of forming the openings 50 and 51 and the holes 25a and 25b in the above-described steps can be performed in a different order from the above-described steps. For example, after the first disk 34 and the second disk 35 are bonded together, the openings 50 and 51 and the hole 25 (the state in which the hole 25a and the hole 25b are connected) may be formed.

  The following manufacturing method may be used instead of the process of manufacturing the first disk 34 in steps 101 to 109 described above. Silicone resin is molded into a CD shape, and the first disk 34 is produced by penetrating each position of the storage unit 41, the analysis unit 46, the waste liquid unit 47, and the flow path 48 using a laser beam or the like. Thereafter, the divided pieces constituting the first disk 34 may be fixed to the second disk 35. Alternatively, only the storage section 41 and the like, the analysis section 46 and the waste liquid section 47 may be made into a penetrating region using a laser beam or the like, and the flow path 48 may be processed into a concave shape. Further, each position of the storage unit 41, the analysis unit 46, the waste liquid unit 47, and the flow path 48 may be processed into a concave shape using a laser or the like.

<4. Liquid feeding principle of CD type microchip>
FIG. 7 is a view for explaining the liquid feeding principle using the CD type microchip of FIG.

  After the solutions 71, 72, and 73 are put in the storage portions 41, 42, and 43 located at different distances from the center of the CD-type microchip 2, respectively, the CD-type microchip 2 is rotated, and the number of rotations is increased. As a result, the solutions 71, 72, and 73 can be guided to the analysis unit 46 in the order of the storage unit 42 farthest from the center of the CD-type microchip 2, the storage unit 42 next to the next, and the storage unit 43 closest to the next. The principle of liquid feeding using such centrifugal force F will be described below.

When the CD-type microchip 2 is rotated at an angular velocity omega, the centrifugal force _{F c} applied to the reservoir 41, 42, 43 from the center at a distance R of the CD type microchip 2, be represented by the following formula (1) Can do.
F _c = Rω ² ρπr ² h (1)
Here, r is a circle-converted radius when it is assumed that the shape of the outlets of the storage portions 41, 42, and 43 is an arc that forms part of a certain circle. ρ is the density of the solutions 71, 72, 73. h is the depth of the solutions 71, 72, 73 in the centrifugal direction.
On the other hand, the force F _s for retaining the solutions 71, 72, 73 caused by the surface tension in the reservoirs 41, 42, 43 can be expressed by the following equation (2).
F _s = 2πrγ cos θ Equation (2)
Here, θ is a contact angle. γ is the surface tension.
When F _c > F _s , the solutions 71, 72, and 73 flow out from the storage portions 41, 42, and 43 toward the outlet, and the angular velocity ω at this time can be expressed by Expression (3).
ω = (2γ | cos θ | / Rρrh) ^1/2 Formula (3)
Therefore, the rotation frequency f (unit: rpm) at this time can be expressed by Expression (4).
f = (60 / 2π) · (2γ | cos θ | / Rρrh) ^1/2 Formula (4)

  As apparent from the equation (4), when the distance (R) from the center of the CD microchip 2 to the storage parts 41, 42, 43 is different, the solutions 71, 72, 73 are transferred from the storage parts 41, 42, 43. The number of rotations flowing out is also different. Therefore, by increasing the rotational speed stepwise, the solution can be sent to the analysis unit 46 in order from the storage unit 41 farthest from the center.

<5. Configuration of organic EL>
FIG. 8 shows a perspective view of the organic EL of FIG.

The organic EL 13 has a laminated structure in which five narrow electrodes 82 are disposed on a transparent substrate 81 such as glass, an organic layer 83 is disposed on the four electrodes 82, and an electrode 84 is disposed thereon. The electrode 82 is preferably a transparent electrode excellent in conductivity, and particularly preferably an electrode made of an ITO film. The electrode 84 is preferably a metal layer, particularly preferably an electrode made of an aluminum thin film. In this embodiment, the organic layer 83 is formed by sequentially stacking MoO ₃ , TPD, Ir (ppy) ₃ : CBP, Bphen, Alq ₃ , and LiF, but the stacked structure is merely an example, and the cathode and By applying a voltage between the anodes, electrons and holes are injected from each, and the injected electrons and holes are combined in the light emitting layer to excite the light emitting material of the light emitting layer. Any combination of layers may be used as long as light can be generated upon returning to the ground state.

  Hereinafter, a preferable example of producing the organic EL 13 will be described.

  First, an ITO electrode is prepared. A glass substrate on which ITO is uniformly formed is cut into a 30 mm square with a glass cutter and ultrasonically cleaned using an ultrasonic cleaner in the order of deer clean LX-II, pure water, and IPA. After drying the substrate in an incubator at about 70 ° C., a positive photoresist PMER is spin-coated on the ITO surface of the substrate in a dark room, left for about 20 minutes, and then about 15 on a hot plate at about 85 ° C. Bake for a minute. Next, a photomask on which an ITO electrode pattern is drawn is placed on the substrate, fixed with a vacuum clamp, and then exposed for about 3 minutes using an exposure machine. Next, the substrate is developed by immersing it in a developing solution, sufficiently rinsed with pure water, and then baked for about 15 minutes on a hot plate at about 85 ° C. Next, this substrate was immersed in hydroiodic acid (concentration 55.0 to 58.0%) for about 30 minutes to remove the ITO in the resist-free portion, rinsed thoroughly with pure water, and then washed with acetone and IPA. To remove the resist. Next, ultrasonic cleaning is performed for about 10 minutes each in the order of deer clean, pure water (preferably twice), acetone, and IPA, and the substrate is immersed in new IPA and stored at room temperature.

Next, an organic material and a metal material are set in the organic thin film deposition apparatus. Next, the ITO electrode substrate immersed in IPA and boiled and washed on a hot plate at about 200 ° C. is set on the substrate holder and carried into the glove box. This is transferred from the glove box to the organic vapor deposition chamber of the organic thin film vapor deposition apparatus and evacuated. After confirming that the pressure in the apparatus is 1.0 × 10 ⁻⁴ Pa or less, the substrate holder is set on a mask at the top of the organic chamber, and MoO ₃ , TPD, 6 wt% -Ir (ppy) ₃ : CBP , Bphen, Alq ₃ , LiF in this order. Thereafter, the substrate holder is transferred from the organic vapor deposition chamber to the metal vapor deposition chamber, set on a mask at the top of the metal vapor deposition chamber, and Al is vapor deposited. After all vapor deposition is completed, the atmospheric pressure in the vapor deposition chamber is returned to atmospheric pressure, and the substrate holder is carried into the glove box.

  Next, the organic EL 13 produced in the above process is sealed. A desiccant is attached to the sealing glass on which the ultraviolet prevention film is pasted, and the resulting organic EL 13 is pasted using an ultraviolet curable resin. The UV curable resin is cured by irradiating the device with black light in the glove box for about 2 minutes. Thus, the sealing process is completed.

<6. Analysis method>
FIG. 9 shows a typical flow of analysis using the analyzer of FIG. 1. One is a method (9A) in which the analysis is performed after rotating the CD-type microchip and stopping the rotation. Shows a method (9B) for performing analysis while rotating a CD-type microchip.

(1) Flow of analysis method after sample supply Step 201 (Sample supply step)
As shown in (9A), first, a sample or the like is supplied to the storage portion 41 or the like of the CD type microchip 2. Here, the sample or the like is interpreted to mean “at least the sample”, and may include only the sample or other types of solutions such as a cleaning solution in addition to the sample.

Step 202
Next, the opening part 50 such as the storage part 41 containing the sample or the like is covered with a film or the like. Here, you may cover the opening part 51 of the waste liquid part 47 simultaneously. The cover means such as a film may be a dedicated cover for the storage unit 41 or the like, or a cover for each block 30, or a cover that covers the entire surface of the CD type microchip 2. Note that this step is not an essential step and can be omitted. For example, when a solution is injected into the storage unit 41 or the like using an injection needle or the like, the opening 50 is very small, so that the solution may not easily jump out during analysis. In such a case, the opening 50 may not necessarily be blocked.

Step 203 (Liquid feeding step)
Next, the CD type microchip 2 is set on the rotating shaft 11 connected to the motor 10, and the motor 10 is rotated. When there is only one reservoir 41 or the like, liquid feeding is performed by a single operation. However, when the CD type microchip 2 is provided with five storage portions 41 and the like at different radial distances, in this step, the storage portion 41 on the radially outer side is changed while changing the rotation speed of the motor 10 to five levels. Then, liquid feeding is started, and then liquid feeding is performed from the radially inner storage section 42. Specifically, the motor 10 is rotated at a predetermined number of revolutions to supply liquid from the storage part 41 radially outermost, and then the number of rotations is increased to supply liquid from the storage part 42 radially inward. The number of revolutions is further increased, liquid is fed from the storage part 43 on the inner side in the radial direction, the number of revolutions is further increased, liquid is fed from the storage part 44 on the inner side in the radial direction, and the number of revolutions is further increased. Liquid is fed from the reservoir 45 located radially inside.

Step 204 (Rotation stop step)
The liquid feeding necessary for the analysis is terminated, and it is confirmed that the analysis unit 46 has a sample for analysis, the motor 10 is turned off, and the rotation of the CD microchip 2 is stopped.

Step 205 (lighting step)
Next, the organic EL 13 is caused to emit light and illuminate the analysis unit 46.

Step 206 (measurement step)
Next, the fluorescence derived from the sample generated as a result of illuminating the sample or the like is received by the light receiving unit 16 and measured.

(2) Flow of analysis method in parallel with sample supply Steps 201 to 203 are the same as those described above.

Step 210 (illumination step)
Following step 203, the organic EL 13 is caused to emit light while rotating the CD type microchip 2, and the sample of the analysis unit 46 is illuminated. In this case, the analysis unit 46 comes to the position of the organic EL 13 only once per rotation of the CD type microchip 2. For this reason, it becomes analysis by intermittent illumination. However, as will be described later, the organic EL 13 may also be rotated in accordance with the rotation of the CD microchip 2 so that the sample of the analysis unit 46 is always illuminated.

Step 211 (measurement step)
Next, the fluorescence derived from the sample generated as a result of illuminating the sample or the like is received by the light receiving unit 16 and measured.

<7. Light emitting device / light receiving device synchronized rotation type analyzer>
(1) Lower light emission type FIG. 10 is a modification of the analyzer of FIG. 1, in which the light emitting device and the light receiving device can be rotated in synchronization with the rotation of the CD microchip, and the light emitting device is a CD type. A schematic diagram (10A) and a simplified configuration diagram (10B) of an analyzer of the type disposed below the microchip are shown.

  The analysis apparatus 1a shown in (10A) is a modification of the analysis apparatus 1 of FIG. 1, and is configured by arranging the light emitting device 4 on a turntable 90 positioned below the CD-type microchip 2. In (10A), the light receiving device 5 is also disposed on the turntable 90, but may be disposed on the opposite side of the turntable 90 with the CD-type microchip 2 interposed therebetween. The analyzer 1a according to this modification is different from the analyzer 1 of FIG. 1 in that the light emitting device 4 and the light receiving device 5 can be rotated in synchronization with the rotation of the CD microchip 2.

  The rotating shaft 11 and the motor 10 are driving units that rotationally drive the CD-type microchip 2 and constitute the main part of the first rotating mechanism 3. The turntable 90, the rotation shaft 11, and the motor 10 are drive units that enable the organic EL 13 to rotate in synchronization with the CD type microchip 2, and constitute a main part of the second rotation mechanism. At the same time, the turntable 90, the rotating shaft 11, and the motor 10 are driving units that allow the light receiving unit 16 to rotate in synchronization with the CD-type microchip 2, and also constitute the main part of the third rotating mechanism. . Here, the rotating shaft 11 and the motor 10 are drive units common to the first rotating mechanism, the second rotating mechanism, and the third rotating mechanism. That is, here, the first rotation mechanism, the second rotation mechanism, and the third rotation mechanism have a common drive unit. However, a motor common to each of the first rotation mechanism, the second rotation mechanism, and the third rotation mechanism or two of them and a rotation shaft connected thereto may be provided. Further, the turntable 90 is a component common to the second rotation mechanism and the third rotation mechanism, but the second rotation mechanism and the third rotation mechanism may include separate turntables. Further, the entire light emitting device 4 is not limited to being disposed on the turntable 90, and any element in the components of the light emitting device 4 may be disposed on the turntable 90 as long as at least the organic EL 13 is disposed on the turntable 90. good. Similarly, the entire light receiving device 5 is not limited to being arranged on the turntable 90, and as long as at least the light receiving unit 16 is arranged on the turntable 90, which elements of the light receiving device 5 are arranged on the turntable 90. Also good.

  Further, in the present application, “synchronized and rotatable” means that at least two rotation objects can be rotated at the same number of rotations regardless of whether the number of rotations can be changed. In the analyzer 1a shown in (10A), the CD type microchip 2 and the turntable 90 are connected to one rotating shaft 11, so that the analyzing unit 46, the organic EL 13 and the light receiving unit 16 are connected to the rotating shaft 11. By fixing at the same distance, the analysis unit 46, the organic EL 13 and the light receiving unit 16 can be analyzed under a condition in which relative displacement does not occur at both the radial direction and the length direction from the rotating shaft 11. As a result, analysis with high reproducibility becomes possible. In addition, by adopting a configuration in which the CD-type microchip 2, the organic EL 13 and the light receiving unit 16 can be rotated together, complicated wiring can be eliminated and the entire apparatus can be made compact. Furthermore, since the analysis can be performed while rotating the CD microchip 2, the reaction process can be measured in real time and the operation can be automated. When the measurement is completed, the next measurement can be started only by replacing the CD-type microchip 2. Further, since the CD type microchip 2 is arranged on the turntable 90 on which the organic EL 13 is arranged and analysis is performed, the setting of the CD type microchip 2 becomes extremely simple.

  (10B) is a block diagram for more easily explaining the configuration of the analyzer 1a of (10A), and also shows the configuration omitted in (10A). The dichroic mirror 91 is a component that changes the direction of light from the organic EL 13 to the hemispherical lens 15 side. The amplifier 92 is connected to the light receiving unit 16 and is a component that amplifies a signal from the light receiving unit 16. The data processor 93 is a component that performs analysis or data processing before analysis based on the signal amplified by the amplifier 92. The memory 94 is a component that can store data processed by the data processor 93, and may be a card-type memory chip that can be easily detached from the turntable 90. The battery 95 is a detachable power supply source common to the light emitting device 4 and the light receiving device 5. When a commercial power source is used, the light emitting device 4 and the light receiving device 5 can be driven from the outside via the power source 12 without using the battery 95.

  When the analyzer 1 a having such a configuration is used, the light reflected from the organic EL 13 by the dichroic mirror 91 and collected through the hemispherical lens 15 is illuminated onto the sample in the analysis unit 46. The sample receives this light and fluoresces. Light from the sample enters the light receiving unit 16 typified by a CCD camera or the like and is amplified by an amplifier 92. The data processor 93 performs data processing based on the amplified optical signal.

(2) Upper light emission type FIG. 11 is a modified example different from the analysis device of FIG. 10, in which the light emitting device and the light receiving device can be rotated in synchronization with the rotation of the CD type microchip, and the organic EL A schematic diagram (11A) and a simplified configuration diagram (11B) of an analyzer of the type disposed above a CD-type microchip are shown.

  The analysis apparatus 1b shown in (11A) is a further modification different from the analysis apparatus 1a in (10A), and includes an organic EL 13 arranged above the CD-type microchip 2. In (11A), the light receiving unit 16 is disposed below the CD-type microchip 2, but may be disposed on the same side as the organic EL 13. Similar to the analyzer 1a, the analyzer 1b according to this modification is different from the analyzer 1 of FIG. 1 in that the light emitting device 4 and the light receiving device 5 can be rotated in synchronization with the rotation of the CD type microchip 2. .

  Most of the components of the light emitting device 4 except for the power source 12 are arranged on a first turntable 96 above the CD type microchip 2. On the other hand, all the components of the light receiving device 5 including the power source 12 are arranged on the second turntable 97 below the CD type microchip 2. The first turntable 96, the rotation shaft 11 and the motor 10 are drive units that enable the organic EL 13 to rotate in synchronization with the CD type microchip 2, and constitute the main part of the second rotation mechanism. The second turntable 97, the rotating shaft 11, and the motor 10 are driving units that allow the light receiving unit 16 to rotate in synchronization with the CD-type microchip 2, and constitute the main part of the third rotating mechanism. The rotating shaft 11 and the motor 10 are drive units common to the first rotating mechanism, the second rotating mechanism, and the third rotating mechanism. That is, here, the first rotation mechanism, the second rotation mechanism, and the third rotation mechanism have a common drive unit. However, a motor common to each of the first rotation mechanism, the second rotation mechanism, and the third rotation mechanism or two of them and a rotation shaft connected thereto may be provided. Further, the entire light emitting device 4 may be disposed on the first turntable 96. Alternatively, the power supply 12 may be disposed on the first turntable 96 and a part of the light receiving device 5 including at least the light receiving unit 16 may be disposed on the second turntable 97.

  In the analyzer 1b, the CD type microchip 2, the first turntable 96, and the second turntable 97 are connected to one rotating shaft 11, so that the analyzer 46, the organic substance, and the organic turntable are the same as the analyzer 1a. By fixing the EL 13 and the light receiving unit 16 at the same distance from the rotating shaft 11, the analysis unit 46, the organic EL 13 and the light receiving unit 16 are relatively displaced in both the radial direction and the length direction from the rotating shaft 11. Analysis is possible under conditions that do not occur. As a result, analysis with high reproducibility becomes possible. In addition, by adopting a configuration in which the CD-type microchip 2, the organic EL 13 and the light receiving unit 16 can be rotated together, complicated wiring can be eliminated and the entire apparatus can be made compact. Furthermore, since the analysis can be performed while rotating the CD microchip 2, the reaction process can be measured in real time and the operation can be automated. When the measurement is completed, the next measurement can be started only by replacing the CD-type microchip 2.

  (11B) is a block diagram for more easily explaining the configuration of the analyzer 1b of (11A), and also shows a configuration omitted in (11A). Unlike the analyzer 1a, the analyzer 1b has the light receiving unit 16 and the organic EL 13 arranged opposite to each other with the analyzer 46 interposed therebetween. Other components of the analyzer 1b are the same as those of the analyzer 1a. When the analyzer 1b having such a configuration is used, the light from the organic EL 13 can be condensed by the hemispherical lens 15 and can be illuminated onto the sample in the analysis unit 46. The sample receives this light and fluoresces. Light from the sample enters the light receiving unit 16 typified by a CCD camera or the like and is amplified by an amplifier 92. The data processor 93 performs data processing based on the amplified optical signal.

<8. Performance Evaluation of Fluorescence Detection System Using CD Type Microchip>
FIG. 12 shows a condition (12A) for introducing a fluorescent reagent by changing the concentration and volume in five reservoirs formed in one block in the CD-type microchip in FIG. 3, and fluorescence detection under that condition. The result (12B) which performed the performance evaluation of the system is shown.

Using resorufin, which is a fluorescent reagent, as a model reagent, performance evaluation of a fluorescence detection system using the analyzer 1 including the organic EL 13 and the CD microchip 2 was performed. Storage part 41 (referred to as “No. 1”), storage part 42 (referred to as “No. 2”), storage part 43 (referred to as “No. 3”), storage part 44 (referred to as “No. 4”) of the CD-type microchip 2. ) And the reservoir 45 (referred to as “No. 5”) were introduced with resorufin aqueous solutions having different concentrations and volumes. Specifically, 25 μL of 10 μM resorufin aqueous solution is introduced into the reservoir 41, 20 μL of 20 μM resorufin aqueous solution is introduced into the reservoir 42, and 15 μL of 40 μM resorufin aqueous solution is introduced into the reservoir 43. Then, 10 μL of a 80 μM resorufin aqueous solution was introduced into the reservoir 44, and 5 μL of a 160 μM resorufin aqueous solution was introduced into the reservoir 45. After the introduction of each aqueous solution, the CD-type microchip 2 was rotated to sequentially introduce resorufin aqueous solutions having different concentrations into the analysis unit 46. Thereafter, the rotation of the CD microchip 2 was stopped, the excitation light collected from the organic EL 13 by the hemispherical lens 15 was applied to the center of the analysis unit 46, and the fluorescence generated thereby was detected by a CCD camera. At this time, the current density of the organic EL 13 was 21 mA / cm ² , and the integration time of the signal from the CCD camera was 500 ms. As a result, the graph (12B) plotting the relationship between the resorufin concentration and the fluorescence intensity showed good linearity with a regression equation y = 134.24X + 1333.5 and a correlation coefficient R ² = 0.993. This means that the concentration of resorufin can be uniquely specified by measuring the fluorescence intensity. Moreover, the detection limit (S / N = 3) of resorufin in this system was estimated to be 9.9 μM.

<9. Immunofluorescence detection method for immunoglobulin A>
FIG. 13 is a diagram for explaining a fluorescence detection method for immunoglobulin A, which is positioned as a stress marker contained in saliva, using the analyzer of FIG. 1. A state (13B) of the analysis unit that changes in accordance with the liquid feeding is shown.

  (13A) shows a situation in the analysis unit 46 immediately before illumination using the organic EL 13 and a reaction formula in which a substrate in the substrate solution 104 reacts with hydrogen peroxide in the presence of HRP to generate a fluorescent substance. An anti-immunoglobulin A antibody (anti-IgA antibody) 100 is adsorbed in advance to the analysis unit 46 of the CD-type microchip 2 (13B (a)), and then blocking is performed on the non-adsorbed portion of the anti-IgA antibody 100. A reagent (hereinafter referred to as “BSA”) 101 is adsorbed (13B (b)), and then in this state, immunoglobulin A (IgA) 102 is introduced into the analysis unit 46, and the anti-IgA antibody 100 and IgA102 are introduced. Then, the HRP-labeled anti-IgA antibody 103 is introduced into the analyzer 46 (13B (d)). Finally, the substrate solution 104 is sent to the analysis unit 46 (13B (e)). As a result, an environment in which IgA102 binds to anti-IgA antibody 100, HRP-labeled anti-IgA antibody 103 binds thereon, and substrate solution 104 exists around it is formed in analysis unit 46. Under such an environment, as shown in the reaction formula (13A), the substrate in the substrate solution 104 reacts with hydrogen peroxide in the presence of HRP to generate a fluorescent substance. When excitation light from the organic EL 13 is illuminated onto the analysis unit 46, fluorescence resulting from the presence of the fluorescent material is emitted from the analysis unit 46. Here, the anti-IgA antibody is an antibody that specifically binds to IgA, and in this antigen-antibody reaction, the anti-IgA antibody is an antibody and IgA is an antigen.

<10. Measurement of immunoglobulin A>
FIG. 14 shows the measurement method (14A) and measurement result (14B) of immunoglobulin A using the analyzer of FIG.

The following method relates to detection of a substance based on specific binding between an antigen and an antibody in the analysis unit 46. The anti-IgA antibody 100 was immobilized on the inner wall of the analysis unit 46 in advance, and a blocking process was performed with BSA101. At this stage, at least an antibody capable of binding to the antigen in the sample is arranged in the analysis unit 46. In addition, although the blocking process using BSA101 is a process required in order to prevent the nonspecific adsorption | suction of the coexisting protein contained in a sample, it is not an essential process. Next, the following liquids 1), 2), 3), 4) and 5) were introduced into the reservoir 41, the reservoir 42, the reservoir 43, the reservoir 44 and the reservoir 45 of the CD microchip 2. .
1) 25 μL of 0-1800 ng / mL IgA solution (solution containing IgA102) diluted with 0.1 M phosphate buffer (pH = 7.4)
2) Tris-HCl buffer (washing solution 110) 20 μL
3) 15 μL of 5 μg / mL HRP-labeled anti-IgA antibody solution (solution containing HRP-labeled anti-IgA antibody 103) diluted with Tris-HCl buffer
4) 10 μL of Tris-HCl buffer (washing solution 110)
5) 5 μL of 0.1 mM Amplex® Red solution (substrate solution 104) containing 1 mM H ₂ O ₂ diluted with 0.1 M phosphate buffer (pH = 7.4)

Next, a sealing film for microplate (1-6774-02, manufactured by ASONE Co., Ltd.) was pasted on the CD-type microchip 2, and all the storage portions 41 and the like were sealed. Next, the CD microchip 2 was rotated, and the number of rotations was increased stepwise so that each solution in the storage unit 41 and the like was sequentially introduced into the analysis unit 46. In addition, about 10 minutes after introducing the solution containing IgA102 and the solution containing the HRP-labeled anti-IgA antibody 103 into the analysis unit 46, and about 3 minutes after introducing the substrate solution 104, respectively, the CD type The microchip 2 was allowed to stand to secure reaction time for antigen-antibody reaction and enzyme-substrate reaction. The fluorescence intensity of the fluorescent substance 111 (resorufin) produced by the enzyme-substrate reaction was measured using the analyzer 1 equipped with the organic EL 13. At this time, the current density of the organic EL 13 was 21 mA / cm ² , and the integration time of the signal from the light receiving unit (CCD camera) 16 was 500 ms.

  As shown in the measurement result (14B) of the fluorescence intensity, it was found that a calibration curve in which the fluorescence intensity increases as the concentration of IgA102 increases can be obtained. The limit of detection of IgA (S / N = 3) in this system was estimated at 35.9 ng / mL. It is known that about 110 to 220 μg / mL of IgA is contained in human saliva, and its concentration increases due to stress. Therefore, it is considered that an objective human stress evaluation can be performed by using an analysis system using the analyzers 1, 1a, 1b.

  The conventionally known ELISA method using a 96-well microplate requires complicated operations such as addition of reagents and washing. However, in the present embodiment, since the measurement can be performed simply by setting the sample solution on the CD type microchip 2 and rotating it, the operation becomes very simple. Further, in the conventional ELISA method, the reaction of anti-IgA and IgA requires 60 minutes, the reaction of IgA and HRP-labeled anti-IgA requires 60 minutes, and the enzyme reaction requires 150 minutes in total. However, in the method using CD type microchip 2, measurement is possible in 23 minutes in total, 10 minutes for reaction of anti-IgA and IgA, 10 minutes for reaction of IgA and HRP-labeled anti-IgA, and 3 minutes for enzyme reaction. Was shortened to about 1/6. The size and weight of the fluorescence analysis system using the organic EL 13 are about 200 mm × 275 mm × 100 mm and 1.9 kg, respectively, and a commercially available microplate reader (manufactured by TECAN, SPECTRA FLUOR, 480 mm × 440 mm × 280 mm, 28 kg) and Compared to this, a significant reduction in size and weight was achieved. Such a system is considered to have the potential to revolutionize on-site environmental measurement and bedside medical testing.

  The present invention can be used for analysis in the environment, food, medical care, quality control, and the like.

1,1a, 1b Analyzer (Analyzer using a compact disk type microchip)
2 CD type microchip (compact disc type microchip)
3 First Rotation Mechanism 4 Light Emitting Device 5 Light Receiving Device 10 Motor (Driver for First Rotation Mechanism, Second Rotation Mechanism, and Third Rotation Mechanism)
11 Rotating shaft (Drive section of first rotating mechanism, second rotating mechanism, and third rotating mechanism)
13 Organic EL (Organic Electroluminescence)
16 Light receiver (CCD camera, etc.)
41, 42, 43, 44, 45 Storage section 46 Analysis section 48 Flow path 90 Turntable (drive section for second rotation mechanism and third rotation mechanism)
96 1st turntable (drive part of 2nd rotation mechanism)
97 Second turntable (drive part of the third rotation mechanism)
100 Anti-IgA antibody (a kind of antibody)
102 IgA (antigen)

Claims (6)

An analyzer that can be used to analyze a sample and can send liquid from the center to the outside in the radial direction using centrifugal force when rotating,
At least one or more reservoirs for storing the sample,
An analysis unit that is arranged on the outer side in the radial direction from the storage unit and serves as an analysis field for the sample sent from the storage unit, and a flow path for liquid connection that connects the storage unit and the analysis unit,
A compact disc type microchip comprising at least
A light-emitting device including organic electroluminescence as a light source that emits light toward the analysis unit;
A light receiving device including a light receiving unit that receives light from the analysis unit;
A first rotation mechanism for rotationally driving the compact disc type microchip;
A second rotation mechanism for rotating the organic electroluminescence,
A third rotation mechanism that rotationally drives the light receiving unit,
The organic electroluminescence and the light receiving portion you can rotate in synchronization with the compact disc microchip analysis device using a compact disc microchip. The analyzer using a compact disk type microchip according to claim 1 , further comprising a drive unit common to the first rotation mechanism, the second rotation mechanism, and the third rotation mechanism. The analyzer using the compact disk type microchip according to claim 1 or 2 , wherein the organic electroluminescence is disposed below the compact disk type microchip. The analyzer using the compact disc type microchip according to claim 1 or 2 , wherein the organic electroluminescence is disposed above the compact disc type microchip. The analyzer using the compact disc type microchip according to any one of claims 1 to 4 , which is used for detecting a substance based on a specific binding between an antigen and an antibody in the analysis unit. 6. The analyzer using a compact disc type microchip according to claim 5 , wherein at least an antibody capable of binding to an antigen in a sample is disposed in the analysis unit.

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