JP2014006084A - Microchip and analyzer using same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a microchip high in detection sensitivity and detection accuracy of scattered light, and an analyzer using the same.SOLUTION: A microchip comprises a chamber 5 having at least a first light transmissive substrate 1 and a second substrate 2 for reflecting at least a part of light incident in the chamber 5 via the first substrate 1 toward the inside of the chamber 5.

Description

  The present invention relates to a microchip and an analyzer using the microchip.

  In recent years, various measuring devices based on scattered light generated when various particles are irradiated with light have been actively developed.

  For example, as a measuring device based on scattered light, a turbidimeter, a latex agglutination reaction measuring device, and the like can be cited (for example, see Patent Document 1 and Patent Document 2).

  The various devices described above are basically designed on the same principle. The principle of the various measuring apparatuses described above will be described with reference to FIG.

  As shown in FIG. 9, in the prior art, particles 103 are introduced into a space between the substrate 101 and the substrate 102. Since it is necessary to transmit light, both the substrate 101 and the substrate 102 are formed of a light transmissive substrate.

  Light is irradiated to the particle 103 from a light source (not shown), and as a result, backscattered light and forward scattered light traveling in different directions are generated from the particle 103.

  The backscattered light passes through the substrate 101 and scatters outside the space sandwiched between the substrates. On the other hand, the forward scattered light passes through the substrate 102 and is scattered outside the space between the substrates and opposite to the direction in which the backward scattered light is scattered.

  In the prior art, either one of the back scattered light and the forward scattered light is detected by a detector (not shown).

JP 11-230889 A (published August 27, 1999) JP 2002-82118 A (published on March 22, 2002)

  However, the conventional techniques as described above have a problem that the detection sensitivity and detection accuracy of scattered light are low, particularly when detection on a microchip scale is considered.

  As shown in FIG. 9, in the prior art, either back-scattered light or forward-scattered light is detected, so the amount of scattered light that can be detected is small. Therefore, the conventional technique has a problem that the detection sensitivity and detection accuracy of scattered light are low.

  Moreover, as shown in FIG. 9, in the prior art, the distance that the light from the light source passes through the flow path is extremely short. Specifically, in the prior art, the light from the light source passes through the space between the substrates only for the distance from the substrate 101 to the substrate 102. As a result, the total number of particles irradiated with the light is reduced. Therefore, the conventional technique has a problem in that the total amount of scattered light generated is small and the detection sensitivity and detection accuracy of the scattered light are low.

  The present invention has been made in view of the above-described conventional problems, and an object thereof is to provide a microchip with high detection sensitivity and detection accuracy of scattered light, and an analysis apparatus using the microchip. is there.

  In order to solve the above problems, the microchip of the present invention is a microchip provided with a chamber for containing fine particles, and the chamber includes at least a first substrate having optical transparency and the first substrate. And a second substrate that reflects at least part of the light incident into the chamber through the one substrate toward the inside of the chamber.

  According to the above configuration, light entering the chamber through the first substrate is irradiated to the fine particles in the chamber in the process of traveling from the first substrate toward the second substrate. At this time, backscattered light is generated from the fine particles toward the first substrate. Further, according to the above configuration, at least a part of the light incident into the chamber through the first substrate is reflected to the inside of the chamber by the second substrate, and the reflected light is irradiated to the fine particles in the chamber. Is done. At this time, forward scattered light is generated from the fine particles toward the first substrate. That is, according to the said structure, backscattered light and forward scattered light can be generated toward the same side. Then, by simultaneously measuring the back scattered light and the forward scattered light, it is possible to dramatically improve the detection sensitivity and detection accuracy of the scattered light.

  In the microchip of the present invention, it is preferable that the second substrate has a light reflectance greater than a light transmittance.

  According to the above configuration, the light reflected to the inside of the chamber by the second substrate can be increased, and as a result, the forward scattered light generated from the fine particles can be increased.

  In the microchip of the present invention, it is preferable that the second substrate has a specularly reflected light amount larger than a scattered light amount.

  According to the above configuration, the amount of scattered light from the second substrate, which becomes a background in the measurement of scattered light from the fine particles, is reduced, so that the detection sensitivity of the scattered light from the fine particles can be increased.

  In the microchip of the present invention, the second substrate includes a base substrate and a light reflection layer provided on a surface of the surface of the base substrate that is disposed toward the inside of the chamber. Preferably it is formed.

  Since the light reflection layer closer to the space inside the chamber can detect forward scattered light more effectively, according to the above configuration that satisfies the condition, the signal value (detected scattered light by a detector or the like) Signal range) can be increased.

  In the microchip of the present invention, the second substrate includes a base substrate and a light reflection layer provided on a surface of the surface of the base substrate that is disposed toward the outside of the chamber. Preferably, the base substrate is a light transmissive substrate.

  According to the said structure, the base substrate is arrange | positioned toward the space inside a chamber. That is, the base substrate is disposed between the space inside the chamber and the light reflecting layer. Therefore, with the above configuration, the light reflection layer can be protected (for example, surface oxidation prevention, contamination prevention, deterioration prevention, etc.), and thus the light reflection layer can more efficiently reflect light. Thus, the forward scattered light generated from the fine particles can be increased. Note that since the base substrate is a light-transmitting substrate, light can pass through the base substrate, and light after passing through the base substrate can be efficiently reflected by the light reflecting layer.

  In the microchip of the present invention, the first substrate is preferably formed of a light transmissive substrate.

  According to the above configuration, the loss of light when passing through the first substrate can be reduced, so that the signal value can be increased.

  In the microchip of the present invention, the second substrate has a light reflectance in the regular reflection direction of 10% or more and less than 100%, and the intensity of the light in the regular reflection direction is reflected in a direction other than the regular reflection direction. It is preferable that the intensity of light is greater than the intensity of light.

  According to the above configuration, the second substrate (more specifically, the light reflecting layer of the second substrate) can be easily and efficiently produced.

  In the microchip of the present invention, it is preferable that the light reflecting surface of the second substrate is formed of at least a metal.

  Furthermore, in the microchip of the present invention, the metal is preferably at least one selected from the group consisting of titanium, chromium, copper, aluminum, aluminum alloy, silver, platinum, and gold.

  According to the above configuration, the light reflected toward the inside of the chamber by the second substrate can be increased, and as a result, the forward scattered light generated from the fine particles can be increased. Moreover, according to the said structure, a 2nd board | substrate can be produced easily.

  In the microchip of the present invention, it is preferable that the light reflecting surface of the second substrate is formed of at least a dielectric film.

  According to the above configuration, the light reflectance of the second substrate can be increased.

  In the microchip of the present invention, it is preferable that a hole communicating with the chamber for injecting a sample into the space inside the chamber is provided.

  According to the above configuration, various samples including fine particles can be easily injected into the chamber through the holes. Therefore, a desired measurement using various samples can be easily performed.

  In the microchip of the present invention, the holes are preferably for introducing latex particles as the fine particles into the chamber.

  According to the said structure, the microchip of this invention can be used as a microchip (microchip for latex aggregation immunoturbidimetry) for implementing a latex agglutination immunoturbidimetry.

  In the microchip of the present invention, the chamber is connected to the microchannel, and the shape of the cross section of the chamber perpendicular to the liquid feeding direction and the shape of the cross section of the microchannel perpendicular to the liquid feeding direction are: The same shape is preferred.

  According to the said structure, while being able to make the shape of a microchip simple, liquid feeding of a sample can be performed easily.

  In order to solve the above-described problems, the analyzer of the present invention detects a light source for irradiating the first substrate of the microchip of the present invention with light and scattered light generated from the fine particles contained in the chamber. And a detector for doing so.

  According to the said structure, light is irradiated to a 1st board | substrate with a light source, and the said light injects into a chamber through a 1st board | substrate. Then, the forward scattered light and the back scattered light generated by the light incident into the chamber are detected by the detector. Therefore, according to the above configuration, a highly sensitive and highly accurate analyzer using the microchip of the present invention can be realized with a simple configuration.

  In the analyzer of the present invention, it is preferable that the detector is disposed on the same side as the light source with the first substrate of the microchip as a boundary.

  According to the above configuration, the forward scattered light and the back scattered light generated by the light incident on the chamber can be detected by a small number of detectors (for example, one).

  The analyzer according to the present invention includes a reflecting mirror that reflects scattered light generated from the fine particles toward the second substrate, and the detector is opposite to the light source with the first substrate of the microchip as a boundary. It is preferable to arrange on the side.

  According to the said structure, the highly sensitive and highly accurate analyzer using the microchip of this invention is realizable by simple structure.

  The analyzer according to the present invention preferably includes at least one of an optical slit and a condensing lens disposed between the light source and the first substrate of the microchip.

  According to the above configuration, light can be efficiently applied to the fine particles in the chamber, and as a result, the detection sensitivity and detection accuracy of the scattered light can be further improved.

  In the analyzer of the present invention, the detector is provided at a position that does not receive light in the regular reflection direction of the second substrate of the microchip out of the light emitted from the light source to the microchip. It is preferable.

  According to the above configuration, it is possible to prevent the detector from detecting light other than the forward scattered light and the back scattered light generated from the fine particles. That is, the background during measurement can be kept low. As a result, the detection sensitivity and detection accuracy of scattered light can be further improved.

  In the analyzer of the present invention, when the scattered light of the microchip that does not contain the fine particles is detected by the detector, the detection value is the intensity of light that the light source irradiates toward the first substrate. It is preferable that a determination unit for determining whether or not it is 5% or less is provided.

  According to the above configuration, it is possible to determine whether or not the background during measurement is low, in other words, whether or not measurement can be performed with high detection sensitivity and high detection accuracy. Specifically, when the scattered light of the microchip that does not contain the fine particles is detected by the detector, the detection value is 5% or less of the intensity of the light emitted from the light source toward the first substrate. Therefore, it can be determined that the background during measurement is low (in other words, measurement can be performed with high detection sensitivity and high detection accuracy). Examples of factors that increase the background during measurement include the position of the detector and the quality of the microchip. Therefore, when the determination unit determines that the background during measurement is high, it is possible to adjust the position of the detector and replace the microchip used for measurement.

  In the analyzer of the present invention, the light source is such that the irradiation area of the light incident into the chamber in a direction perpendicular to the liquid feeding direction does not overlap with the wall surfaces other than the first substrate and the second substrate. It is preferable.

  According to the said structure, the light which injected into the chamber through the 1st board | substrate is not reflected by structures other than a 2nd board | substrate. That is, according to the above configuration, it is easy to control the direction in which light incident in the chamber is reflected (for example, limited to one direction). As a result, the background at the time of measurement generated by the detector detecting light other than the forward scattered light and the back scattered light generated from the fine particles can be kept low.

  Since the present invention enables detection of scattered light using a microchip, the scattered light can be measured more easily and inexpensively.

  Since the present invention can simultaneously detect forward scattering and backscattering, it is possible to improve the detection sensitivity and detection accuracy of scattered light.

  The present invention can maintain a long optical path length of light passing through a microchip even if the microchip is miniaturized (for example, thinned). That is, according to the present invention, the microchip can be reduced in size (for example, reduced in thickness) while maintaining high detection sensitivity and detection accuracy of scattered light.

  The present invention can realize simple measurement with a microchip based on various measurement methods (for example, latex agglutination immunoturbidimetry, turbidimeter, particle counter, etc.).

It is a schematic diagram which shows one Embodiment of the microchip of this invention. It is the figure which showed the relationship between the regular reflection light by the irradiation light irradiated to the 2nd board | substrate, scattered light, and absorption light in one Embodiment of the microchip of this invention. (A) And (b) is a schematic diagram which shows an example of embodiment of a chamber and a microchannel in the microchip of this invention. (A) And (b) is a schematic diagram which shows one Embodiment of the analyzer of this invention. It is a schematic diagram which shows one Embodiment of the analyzer of this invention. It is a schematic diagram which shows an example of embodiment of a light source in the analyzer of this invention. It is a graph which shows the test result in the Example of this invention. It is a graph which shows the test result in the Example of this invention. It is a schematic diagram which shows the concept of the conventional scattered light measuring apparatus.

  An embodiment of the present invention will be described below with reference to FIGS. 1 to 8, but the present invention is not limited to this.

[1. Microchip)
[1-1. Action and effect of microchip)
First, the operation and effect of the microchip of the present embodiment will be described with reference to FIG.

  The microchip of the present embodiment has a chamber 5 formed by the first substrate 1 and the second substrate 2, and various fine particles 3 can be accommodated in the chamber 5.

  Light from a light source (not shown) enters the chamber 5 through the first substrate 1. Then, light immediately after entering the chamber 5 with respect to the fine particles 3 (in other words, light traveling from the first substrate 1 side to the second substrate 2 side) and the second substrate 2 after entering the chamber 5. (In other words, light traveling from the second substrate 2 side to the first substrate 1 side) is irradiated.

  When the fine particles 3 are irradiated with light immediately after entering the chamber 5, backscattered light directed toward the first substrate 1 is generated. On the other hand, when the fine particles 3 are irradiated with light reflected by the second substrate 2 after entering the chamber 5, forward scattered light directed toward the first substrate 1 is generated.

  Since the microchip of the present embodiment can generate back scattered light and forward scattered light traveling in the same direction at the same time, the scattered light can be detected by detecting the scattered light using, for example, one detector. Sensitivity and detection accuracy can be dramatically increased.

  Further, as shown in FIG. 1, in the microchip of the present embodiment, since the light is reflected by the second substrate 2, the distance that the light passes through the chamber 5 can be increased. In this case, light passes back from the first substrate 1 to the second substrate 2 to generate backscattered light, is reflected, and travels from the second substrate 2 to the first substrate. Scattered light will be generated. That is, the microchip of the present embodiment can drastically increase the total amount of scattered light generated by light of the same intensity as compared with the conventional technique. Also by this, with the microchip of the present embodiment, the detection sensitivity and detection accuracy of scattered light can be dramatically increased.

  Hereinafter, each configuration will be described in detail.

[1-2. Microchip configuration)
The microchip according to the present embodiment includes a chamber 5 for accommodating the fine particles 3.

  The fine particles 3 are not particularly limited as long as they can generate scattered light when irradiated with light. For example, the fine particles 3 may be artificially produced fine particles (for example, metal beads, latex beads, micelles, etc.), or natural fine particles existing in nature (for example, dust or dust that becomes an allergen substance). Or microorganisms). The fine particles 3 may be an aggregate of the same fine particles or an aggregate of different fine particles.

  As described above, latex beads can be used as the fine particles 3. In addition, latex beads whose surface is covered with a desired antibody can be used as the fine particles 3. According to the said structure, the microchip of this Embodiment can be utilized as a microchip for implementing a latex agglutination immunoturbidimetry.

  The chamber 5 includes at least a first substrate 1 having optical transparency, and a second substrate 2 that reflects at least part of light incident into the chamber 5 through the first substrate 1 toward the inside of the chamber 5. And is formed by.

  The shape of each surface (for example, surfaces facing each other) of the first substrate 1 and the second substrate 2 is not particularly limited, and may be a flat surface, a curved surface, or uneven. It may be a flat surface or a curved surface, or any combination thereof.

  The positional relationship between the first substrate 1 and the second substrate 2 is not particularly limited, and the second substrate 2 may be provided at a position where light incident into the chamber through the first substrate 1 can be irradiated.

  For example, it is also possible to provide both so that the first substrate 1 and the second substrate 2 face each other. More specifically, it is possible to provide both the first substrate 1 and the second substrate 2 so that they face each other in parallel. When both are provided so that the first substrate 1 and the second substrate 2 face each other in parallel, more specifically, the surfaces of the first substrate 1 and the second substrate 2 facing each other are parallel to each other. What should I do. Of course, the present invention is not limited to such an arrangement relationship of the first substrate 1 and the second substrate 2.

  The space of the chamber 5 can be formed as a space between the first substrate 1 and the second substrate 2. For example, another member (for example, a third substrate and a fourth substrate) is sandwiched between the first substrate 1 and the second substrate 2, and the first substrate 1 and the second substrate 2 are interposed via the another member. It is also possible to form a space of the chamber 5 by adhering. As said another member, the tape etc. which can be bonded on both sides are preferable. It is also possible to form the space of the chamber 5 by integral processing by injection molding or by directly bonding the first substrate 1 and the second substrate 2 with a microchannel pattern.

  The first substrate 1 only needs to be a substrate (light transmissive substrate) that allows at least a part of light irradiated to the first substrate 1 from a light source or the like to enter the chamber 5. Is not particularly limited.

  For example, the first substrate 1 is at least 50%, preferably at least 60%, more preferably at least 70%, more preferably at least 80%, most preferably light emitted from the light source to the first substrate 1. It may be a substrate that allows at least 90% of the light to enter the chamber 5.

  More specifically, as the first substrate 1, a glass substrate or a light-transmitting resin (for example, PMMA (Poly Methyl Methacrylate), PET (Poly Ethylene Terephthalate), COC (Cyclo Olefin Copolymer), etc.) or the like may be used. Is possible.

  The second substrate 2 may be any substrate that can reflect at least part of the light incident into the chamber 5 through the first substrate 1 toward the inside of the chamber 5, and its specific configuration is particularly limited. Not.

  Here, first, how the light applied to the second substrate 2 is decomposed and absorbed will be described with reference to FIG.

  As shown in FIG. 2, the light applied to the second substrate 2 is divided into specularly reflected light, scattered light (scattered reflection), and transmitted light, and a part of the light is absorbed by the second substrate 2.

The intensity of light emitted to the second substrate 2 is “I _irradiation ”, the intensity of light reflected by the second substrate 2 is “I _reflection ”, and the intensity of scattered light generated on the second substrate 2 _Is “I _scattering ”, the intensity of light transmitted through the second substrate 2 is “I _transmission ”, and the intensity of light absorbed by the second substrate 2 is “I _absorption ”. Holds. That means
I _irradiation = I _reflection + I _scattering + I _transmission + I _absorption (Relational formula 1).

  The specific configuration of the second substrate 2 can be determined based on the parameters and relational expression 1 described above. An example is shown below.

For example, the second substrate 2 may have a light reflectance greater than a light transmittance. In other words, the second substrate 2 may be a substrate that satisfies the condition “I _transmission <I _reflection ”.

In addition, the second substrate 2 may have a greater regular reflection light amount than a scattered light amount. In other words, the second substrate 2 may be a substrate that satisfies the condition of “I _scattering <I _reflection ”.

The second substrate 2 has a light reflectance in the regular reflection direction of 10% or more and less than 100%, and the intensity of light in the regular reflection direction is greater than the intensity of light reflected in other than the regular reflection direction. Can also be large. In other words, the second substrate 2 may be a substrate that satisfies the conditions of “0.1 × I _irradiation ≦ I _reflection <1.0 × I _irradiation ” and “I _scattering + I _transmission + I _absorption <I _reflection ”.

  Any of the above configurations can increase the amount of light reflected by the second substrate. As a result, it is possible to increase the forward scattered light generated as a result of the reflected light being applied to the fine particles in the chamber.

  A substrate that satisfies the above-described conditions can be easily selected by measuring each parameter with a known optical device (for example, a spectrophotometer manufactured by Hitachi High-Technologies Corporation). In addition, at the time of filing this application, the data of each parameter described above is well known (see, for example, Chemical Handbook (edited by the Chemical Society of Japan)). Therefore, it is possible to select a substrate that satisfies the above-described conditions based on the document.

  The said 2nd board | substrate 2 may be formed by one structure, and may be formed by the some structure.

  When the second substrate 2 is formed with a plurality of structures, for example, the second substrate 2 is formed on the base substrate and the surface of the base substrate that is disposed toward the inside of the chamber 5. And a light reflection layer provided on the substrate. In the case of this configuration, at least a part of the light that enters the chamber 5 through the first substrate 1 is reflected toward the inside of the chamber 5 by the light reflecting layer.

  The structure of the base substrate is not particularly limited, and a desired structure can be used as appropriate. As the base substrate, for example, a light transmissive substrate can be used, and a light non-transmissive substrate can also be used. In the case of the present embodiment, the light reflecting layer is arranged toward the space inside the chamber 5 and light can be efficiently reflected by the light reflecting layer, so the specific structure of the base substrate is not particularly limited.

  Further, when the second substrate 2 is formed by a plurality of configurations, for example, the second substrate 2 has a base substrate and a surface of the surface of the base substrate that is disposed toward the outside of the chamber. Further, the base substrate may be a light-transmitting substrate. In the case of this configuration, at least a part of the light that enters the chamber 5 through the first substrate 1 is reflected toward the inside of the chamber 5 by the light reflecting layer.

  In this case, since the light after passing through the base substrate is reflected by the light reflection layer, the base substrate needs to be a light transmissive substrate. The specific configuration of the light transmissive substrate is not particularly limited. For example, a glass substrate or a light transmissive resin (for example, PMMA, PET, COC, or the like) is used in the same manner as the first substrate 1 described above. Is possible.

  In the case where the second substrate 2 is formed by a single configuration, the specific configuration is not particularly limited. For example, a material obtained by processing the material for forming the light reflecting layer into a substrate shape is the second substrate. 2 can be used.

  The light reflecting surface of the second substrate 2 (for example, the surface of the second substrate 2 or the light reflecting layer described above) can be formed of a dielectric film (for example, a dielectric multilayer film) and / or a metal film. Although it does not specifically limit as a metal which comprises the said metal film, For example, it may be at least 1 selected from the group which consists of titanium, chromium, copper, aluminum, an aluminum alloy, silver, platinum, and gold | metal | money. Of course, the present invention is not limited to these.

It is also possible to coat a dielectric film as a reflection-enhancing film on the metal film. According to the above configuration, the reflectance of light can be increased. The No particular limitation is imposed on the material of the dielectric film, for _example, a _{TiO 2, Ta 2 O 3,} Si 2 or MgF _2.

  For example, the second substrate 2 is formed using the above-described metal by a known method (evaporation or sputtering), or at least a part of the surface of the second substrate 2 is coated with the above-described metal, thereby reflecting the light reflecting surface. Can be formed.

  The microchip of the present embodiment may be provided with a hole communicating with the chamber 5 in order to inject a sample into the space in the chamber 5.

  More specifically, the microchip of this embodiment is provided with one or a plurality of microchannels, and at least one of the microchannels has one end connected to the chamber 5 and the other end connected to the other end. It may be connected to the hole.

  If it is the said structure, the various samples containing microparticles | fine-particles (for example, latex particle) etc. can be easily inject | poured in the chamber 5 through the said hole. Therefore, desired measurement using various samples can be easily performed.

  In the case where a plurality of microchannels are provided in the microchip of this embodiment, at least one of the microchannels may have one end connected to the chamber 5 and another end connected to a hole different from the hole described above. Good. If it is the said structure, the said another hole can be utilized as a hole for discharge | releasing a sample out of the chamber 5. FIG.

  If both the hole for injecting the sample into the space in the chamber 5 and the hole for discharging the sample to the outside of the chamber are provided, the sample can be continuously extracted using the microchip of this embodiment. Can be measured.

  The chamber 5 and the microchannel may have the same or different cross-sectional shapes in a direction perpendicular to the liquid feeding direction.

  3A, the microchip 10 in the case where the cross-sectional shape in the direction orthogonal to the liquid feeding direction of the chamber 5 and the microchannel 6 (the direction from the left to the right in the paper surface of FIG. 3A) is different, An example of the situation when viewed from above is shown.

  The microchip 10 shown in FIG. 3A is formed so that the cross-sectional area in the direction perpendicular to the liquid feeding direction of the chamber 5 is larger than the cross-sectional area in the direction perpendicular to the liquid feeding direction of the microchannel 6. Has been. Although not shown, the microchip 10 is formed so that the cross-sectional area in the direction perpendicular to the liquid feeding direction of the chamber 5 is smaller than the cross-sectional area in the direction perpendicular to the liquid feeding direction of the microchannel 6. It is also possible to form.

  In FIG. 3B, the microchip 10 in the case where the cross-sectional shape in the direction orthogonal to the liquid feeding direction of the chamber 5 and the microchannel 6 (the direction from the left to the right in the paper surface of FIG. An example of the state when viewed from above is shown. In this case, a partial region of the microchannel 6 functions as the chamber 5.

  According to the above configuration, since the complexity of the configuration of the microchip 10 is reduced, the microchip 10 can be manufactured easily and at low cost.

  In FIGS. 3A and 3B, the chamber 5 and the microchannel 6 are described as separate structures, but these structures can be formed as a single structure. For example, the microchannel 6 and the chamber 5 illustrated in FIG. 3B are configured as a single chamber 5 and the chamber 5 has both a function as a chamber and a function as a microchannel. Is also possible. In this case, the entire upper side of the microchannel 6 and the chamber 5 in FIG. 3B may be the first substrate, and the entire lower side of the microchannel 6 and the chamber 5 in FIG. 3B may be the second substrate. The same applies to the microchannel 6 and the chamber 5 described in FIG.

[2. Analysis equipment〕
The analyzer of the present embodiment is a light source for irradiating light to the first substrate of the microchip of the present invention, and for detecting scattered light generated from fine particles contained in the chamber of the microchip of the present invention. And a detector.

  Since the microchip of the present invention has already been described, the description thereof is omitted here.

  As shown in FIG. 4A and FIG. 4B, the analyzer 20 of the present embodiment can include an insertion portion 15 for inserting the microchip 10. One insertion section 15 may be provided for the analyzer 20, or a plurality of insertion sections 15 may be provided.

  It is preferable that the insertion portion 15 can attach and detach the microchip 10 to and from the analyzer 20. According to the said structure, a desired sample can be measured simply and rapidly. In particular, when there are a plurality of samples, the measurement time can be dramatically shortened. Further, according to the above configuration, when the microchip 10 loses desired performance due to deterioration or the like, the microchip 10 can be easily replaced, so that the detection sensitivity and accuracy of measurement can be maintained high. it can.

  A specific configuration of the light source is not particularly limited, and a known light source can be used as appropriate. In addition, the number of light sources provided in the analyzer according to the present embodiment is not particularly limited, and may be one or plural.

  The analyzer according to the present embodiment may include at least one of an optical slit and a condenser lens arranged between the light source and the first substrate of the microchip.

  According to the above configuration, it is possible to irradiate the microchip with light having high intensity, and thus it is possible to generate stronger scattered light from the fine particles. As a result, the detection sensitivity and detection accuracy of scattered light can be improved.

  Moreover, according to the said structure, light can be irradiated to the local of a microchip. More specifically, only the chamber can be irradiated with light. And since generation | occurrence | production of the reflected light other than a 2nd board | substrate can be suppressed, as a result, it becomes easy to prevent that reflected light is directly detected by a detector. As a result, the detection sensitivity and detection accuracy of scattered light can be improved.

  The configuration of the optical slit and the condensing lens is not particularly limited, and a known optical slit and condensing lens can be used as appropriate.

  Although an example of arrangement | positioning of the optical slit 16 and the condensing lens 17 in the analyzer of this embodiment is shown in FIG. 5, this invention is not limited to this.

  As shown in FIG. 5, in the analyzer of this embodiment, an optical slit 16 and a condenser lens 17 are arranged between a light source 18 and a microchip (chamber 5). More specifically, the optical slit 16 is disposed closer to the light source 18, and the condenser lens 17 is disposed closer to the microchip. The light from the light source 18 enters the condenser lens 17 after passing through the optical slit 16. And the light which passed the said condensing lens 17 is irradiated to a microchip (chamber 5), and, thereby, many scattered light (back scattered light and forward scattered light) is emitted from microparticles | fine-particles.

  The position of the focal point of the light condensed by the condensing lens 17 is not particularly limited as long as it is in the space in the chamber 5. However, it is necessary to make the focus position constant every time. This is because the signal value increases or decreases because the effect of reflecting the irradiation light changes depending on the focus position. For example, the position of the focal point is the first substrate surface in the chamber 5 or the second substrate surface in the chamber 5. Of these focal points, the second substrate surface is preferred. The reason is that the closer the focusing position is to the reflecting surface, the greater the effect.

  The light source 18 travels in a direction in which light incident on the chamber 5 travels directly in the liquid feeding direction (the direction from the left to the right of the paper in the microchip shown in FIG. 5 (shown by the chamber 5 and the microchannel 6)). It is preferable that the irradiation area does not overlap with the wall surfaces of the chamber 5 other than the first substrate and the second substrate.

  Further, as shown in FIG. 6, in the light source 18, the horizontal component of the light emitted from the light source 18 and the extending direction of the microchannel 6 (in other words, the liquid feeding direction) are substantially parallel. preferable. In FIG. 6, the horizontal direction component is indicated by direction A, and the extension direction of the microchannel 6 is indicated by direction B. In this case, the optical path width of the light irradiated by the light source 18 is preferably shorter than the width of the chamber 5.

  According to the said structure, it can prevent that light is reflected by structures other than the 2nd board | substrate of the chamber 5. FIG.

  The detector is not particularly limited as long as it can detect scattered light. As the detector, a known detector may be used as appropriate.

  The detector may be disposed on the same side as the light source with the first substrate of the microchip as a boundary. The position of the detector may be on the same side as the light source with the first substrate of the microchip as a boundary, and the specific position is not particularly limited. Of course, the detector may be arranged on the opposite side of the light source with the first substrate of the microchip as a boundary. In this case, for example, a reflecting mirror may be disposed on the same side as the light source, and a detector that detects light from the reflecting mirror may be disposed on the opposite side of the light source.

  According to the above configuration, the forward scattered light and the back scattered light generated by the light incident on the chamber can be detected simultaneously by a small number of detectors (for example, one). That is, according to the above configuration, the configuration of the analyzer according to the present embodiment can be simplified.

  The detector may be provided at a position that does not receive light in the specular reflection direction of the second substrate of the microchip out of the light emitted from the light source to the microchip. Generally, when the angle between the light beam of the light source and the irradiation surface in the chamber is θ, the position of the detector can take an angle other than 180−θ.

  Whether or not the detector is provided at the above-described position is determined by, for example, detecting the scattered light of the microchip that does not contain the fine particles by the detector, and the detection value is emitted from the light source toward the first substrate. It can be confirmed by determining whether or not the light intensity is 5% or less, more preferably 3% or less, more preferably 1% or less, and most preferably 0%.

  In other words, when the scattered light of the microchip that does not contain the fine particles is detected by the detector, the detection value is 5% or less, more preferably 3% or less, of the intensity of the light emitted from the light source toward the first substrate. If it is more preferably 1% or less, most preferably 0%, it can be determined that the detector is provided at the position described above.

  Therefore, in the analyzer of the present embodiment, when the scattered light of the microchip that does not contain the fine particles is detected by the detector, the detected value is emitted from the light source toward the first substrate. A determination unit for determining whether the light intensity is 5% or less, 3% or less, 1% or less, or 0%. It is preferable to provide.

  The configuration of the determination unit is not particularly limited, and a known configuration can be used as appropriate. Further, the determination unit and the detector can have the same configuration.

<1. Production of Chip (Example)>
As a chip substrate (corresponding to the second substrate), (1) a mirror-finished silicon substrate, (2) a Ti thin film (50 nm) substrate formed on a glass substrate by sputtering, and (3) a thin film on the glass substrate by sputtering. Ti (50 nm) -Au (100 nm) thin film (surface is gold) substrate, (4) Ti (50 nm) -Pt (100 nm) thin film (surface is platinum) substrate formed on a glass substrate by sputtering, (5) Ti (50 nm) -Al (100 nm) thin film (surface is aluminum) substrate formed thin film on glass substrate by sputtering, (6) Ti (50 nm) -Ag (thin film formed on glass substrate by sputtering) A 100 nm) thin film (surface is silver) substrate was used. The (2) Ti thin film substrate had a light transmittance of 20%, but the other substrates did not transmit light.

  Further, a double-sided tape (thickness 50 μm) cut into an I-shape (length 2 cm, width 600 μm) with a laser processing machine (manufactured by GCC) is attached to each of the above-described substrates, and the above-mentioned double-sided tape is attached to the substrate A 1.5 cm square glass substrate (corresponding to the first substrate) was bonded as shown in FIG. 1 to produce a chip in which a flow path was formed between the chip substrate and the glass substrate.

<2. Chip Fabrication (Comparative Example)>
A (0) glass substrate was used as the chip substrate (corresponding to the second substrate). As in the example, a double-sided tape (thickness 50 μm) cut into an I-shape (length 2 cm, width 600 μm) with a laser processing machine (manufactured by GCC) was attached to the glass substrate, and another 1.5 cm square A glass substrate (corresponding to the first substrate) was bonded as shown in the drawing to produce a chip in which a flow path was formed between the glass substrate and the glass substrate.

<3. Evaluation method>
A suspension of latex beads (φ0.1 μm) modified with an anti-CRP monoclonal antibody and a CRP solution (0 mg / dL or 3.5 mg / dL) were mixed at a volume ratio of 1: 1.

  The mixed solution was injected into the prepared chip, and a dark field image was taken 3 minutes after mixing. Since the luminance information of the captured dark field image depends on the size of the scattered light of the latex particles, the luminance information of each chip was compared.

  The above-described dark field image was captured using a dark field microscope (Olympus). However, the light hitting the flow path wall is cut by masking. The wavelength of the irradiation light was 550 ± 50 nm (however, the test result is not influenced by the wavelength of the irradiation light).

<4. Result>
FIG. 7 shows the test results in the case of 0 mg / dLCRP concentration, and FIG. 8 shows the test results in the case of 3.5 mg / dLCRP concentration. When the actual sensitivity (= (luminance at 3.5 mg / dL) − (luminance at 0 mg / dL)) is derived, the sensitivity increases by a factor of 40 when the chip substrate is a silicon substrate. When the Ti substrate is a Ti thin film substrate, the sensitivity is increased 42 times, when the chip substrate is a Ti—Au thin film substrate, the sensitivity is increased 100 times, and when the chip substrate is a Ti—Pt thin film substrate, the sensitivity is increased 180 times. The sensitivity increased by 290 times when the chip substrate was a Ti—Al thin film substrate, and the sensitivity improved by 300 times when the chip substrate was a Ti—Ag thin film substrate.

  The present invention is not limited to the configurations described above, and various modifications can be made within the scope of the claims, and technical means disclosed in different embodiments and examples are appropriately used. Embodiments and examples obtained in combination are also included in the technical scope of the present invention.

  The present invention can be used for a microchip for performing various measurement methods (for example, latex agglutination immunoturbidimetry, turbidimeter, particle counter, etc.). More specifically, the present invention can be used for a turbidimeter or a particle counter.

DESCRIPTION OF SYMBOLS 1 1st board | substrate 2 2nd board | substrate 3 Fine particle 5 Chamber 6 Microchannel 10 Microchip 15 Insertion part 16 Optical slit 17 Condensing lens 18 Light source 20 Analyzer 30 Detector 40 Judgment part 50 Moving apparatus

Claims (20)

A microchip having a chamber for containing fine particles,
The chamber includes at least a first substrate having optical transparency, and a second substrate that reflects at least part of light incident into the chamber through the first substrate toward the inside of the chamber. A microchip characterized by being formed.   The microchip according to claim 1, wherein the second substrate has a light reflectance larger than a light transmittance.   3. The microchip according to claim 1, wherein the second substrate has a specular reflection light amount larger than a scattered light amount.   The second substrate is formed by a base substrate and a light reflection layer provided on a surface of the surface of the base substrate that is disposed toward the inside of the chamber. The microchip according to any one of claims 1 to 3. The second substrate is formed by a base substrate and a light reflecting layer provided on a surface of the surface of the base substrate that is disposed toward the outside of the chamber.
The microchip according to claim 1, wherein the base substrate is a light transmissive substrate.   The microchip according to claim 1, wherein the first substrate is formed of a light transmissive substrate. The second substrate has a reflectance of light in the regular reflection direction of 10% or more and less than 100%,
The microchip according to claim 1, wherein the intensity of light in the regular reflection direction is greater than the intensity of light reflected in other than the regular reflection direction.   The microchip according to claim 1, wherein the light reflecting surface of the second substrate is formed of at least a metal.   9. The microchip according to claim 8, wherein the metal is at least one selected from the group consisting of titanium, chromium, copper, aluminum, aluminum alloy, silver, platinum, and gold.   The microchip according to claim 1, wherein the light reflecting surface of the second substrate is formed of at least a dielectric film.   The microchip according to any one of claims 1 to 10, wherein a hole communicating with the chamber is provided for injecting a sample into a space inside the chamber.   The microchip according to claim 11, wherein the hole is for introducing latex particles as the fine particles into the chamber. The chamber is connected to a microchannel,
13. The micro of claim 1, wherein the shape of the cross section of the chamber perpendicular to the liquid feeding direction and the shape of the cross section of the microchannel perpendicular to the liquid feeding direction are the same shape. Chip. A light source for irradiating light to the first substrate of the microchip according to any one of claims 1 to 13,
And a detector for detecting scattered light generated from the fine particles contained in the chamber.   The analyzer according to claim 14, wherein the detector is arranged on the same side as the light source with the first substrate of the microchip as a boundary. A reflecting mirror for reflecting scattered light generated from the fine particles toward the second substrate;
The analyzer according to claim 14, wherein the detector is disposed on the opposite side of the light source with the first substrate of the microchip as a boundary.   17. The apparatus according to claim 14, further comprising at least one of an optical slit and a condenser lens disposed between the light source and the first substrate of the microchip. Analysis equipment.   The detector is provided at a position that does not receive light in the regular reflection direction of the second substrate of the microchip out of the light emitted from the light source to the microchip. The analyzer according to any one of claims 14 to 17.   Whether the detection value is 5% or less of the intensity of light emitted from the light source toward the first substrate when the detector detects the scattered light of the microchip that does not contain the fine particles. The analyzer according to any one of claims 14 to 18, further comprising a determination unit that determines whether or not.   2. The light source according to claim 1, wherein an irradiation area in a direction perpendicular to the liquid feeding direction of light incident into the chamber does not overlap with a wall surface other than the first substrate and the second substrate. The analyzer according to any one of 14 to 19.

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