JP4673149B2 - Method of using microchip, microchannel, and microchip - Google Patents

Description

  The present invention relates to a method of using a microchip, a microchannel, and a microchip, and relates to a microchannel used for separating, mixing, and detecting a biological material, a method of using a microchip including the same, a microchannel, and Related to microchip.

  2. Description of the Related Art In recent years, a technique called Lab on Chip, which separates, reacts, mixes, measures, and detects biological substances such as blood on a substrate having a size of several centimeters, has attracted attention in the field of medicine and the like. This chip is provided with a micro flow channel of several nm to several mm, and movement, reaction, measurement, and the like of a small amount of sample are realized in the flow channel. Therefore, various measurements and detections can be performed easily in a short time.

  However, since the sample usually contains protein, fats and oils, these may adhere to the wall surface of the microchannel. In addition, bubbles may be generated due to temperature changes during sample operation, and this bubble cannot prevent accurate measurement by blocking the flow of the sample itself or obstructing the detection system. There is a problem.

On the other hand, there is a method of hydrophilizing the wall surface of the microchannel. For example, surface treatment such as corona treatment, plasma treatment, and ultraviolet treatment, and kneading of a hydrophilic substance into a molding material are well known.
However, in the surface treatment, the degree of hydrophilization is weak, and the productivity of the microchip decreases due to an increase in the number of steps. In addition, the kneading of the hydrophilic substance has problems such as deterioration of physical properties such as the microchip itself becoming brittle, loss of the hydrophilic substance from the microchip, and partial hydrophilicity.

Therefore, a method has been proposed in which a hydrophilic polymerizable compound is graft-polymerized on the surface of an uncured energy beam curable resin molded article to make the surface hydrophilic (for example, Patent Document 1). This method is intended to impart hydrophilicity to the microchannel and to prevent the loss of the hydrophilic substance from the microchip.
Japanese Patent Laid-Open No. 10-53658

  However, in such a hydrophilization method, steps such as application of a hydrophilic polymerizable resin, energy irradiation for curing, and removal of an uncured resin must be performed, and the manufacturing process becomes complicated. In addition, since the resin is cured by irradiating light, it is difficult to apply it to a portion that is difficult to irradiate light, such as a side wall of a microchannel.

  The present invention has been made in view of the above problems, and by performing an appropriate treatment such as simple hydrophilization over the entire surface of the microchannel, it is possible to prevent adhesion of a sample and generation of bubbles. An object of the present invention is to provide a microchip usage method, a microchannel, and a microchip capable of performing an accurate analysis with a small amount of sample.

Using the microchip of the present invention, the reagent holding part for holding a reagent, mixing tank, a detector and a method of using at least comprises a microchip micro flow path connecting the mixing tank and the detection unit, ( c) including a step of wetting the detection unit with a mixed solution of a reagent and a sample to be detected by the detection unit before the sample is detected by the detection unit.

  With such a configuration, it is not necessary to perform a special hydrophilization process on the microchip itself as in the prior art, and it is possible to prevent the productivity of the microchip from being lowered, and for the hydrophilization treatment. Without impairing the accompanying physical properties of the microchip, it is possible to prevent the additives and processing agents used in the processing from being mixed into the microchip or the sample or reagent to be measured. In addition, by passing the sample or reagent introduced into the microchip through the detection unit, the contact angle of the mixed solution of the reagent and the sample is as small as about 0 to 10 ° while the detection unit is wet. It is possible to eliminate the limitation due to the shape and dimensions of the detection unit, and all the inner surfaces of the upper surface, the side surface, and the bottom surface of the detection unit can be made hydrophilic simultaneously and uniformly. Furthermore, since the hydrophilic treatment can be performed only on a specific portion by flowing the liquid only on the detection portion, it is possible to sort the hydrophilic portion and the hydrophobic portion at an arbitrary portion. In addition, by reducing the contact angle as described above, adsorption of substances such as proteins and oils and fats to the detection unit surface can be effectively suppressed. At the same time, the generation of air pockets due to fine irregularities on the surface of the detection unit can be suppressed, and as a result, the generation of bubbles with the air pockets as growth nuclei can be suppressed. Analysis can be realized.

In the method of using the microchip of the present invention, (1) the microchip further includes a measuring unit, and further includes (a) a step of measuring a sample, (b) a step of mixing the measured sample with a reagent, (d) the mixture is introduced into the detection unit which has already wetted by the process of wetting the (c) viewed including the steps of detecting the sample, the step of wetting the (c), (b) after step (d) It is performed before the process . Thereby, the effect mentioned above can be realized more reliably.
Further, the microchip further includes a separation unit at the sample introduction port and the separation unit, and further includes (aa) a step of centrifuging the introduced sample. Thereby, only the substance to be measured in the introduced sample can be easily taken out and measured, thereby enabling more accurate detection.

Further, (bb) a step of passing the reagent through the detection unit to wet the detection unit is included. As a result, the detection unit can be wetted with a reagent that does not contain solids, etc., so even if the sample contains solids, it prevents more solids from adhering to the detection unit. In addition, the contact angle of the mixed liquid can be further reduced.
In the method of using the microchip of the present invention, at least one step is performed using centrifugal force. Thereby, the process of wetting the detection unit as described above can be performed very simply, quickly, and more reliably.

  In particular, (b) mixing is performed using a force in a predetermined direction, and (a) weighing, (c) wetting of the detection unit, and (d) introduction into the detection unit are performed in a direction different from the predetermined direction. Or (b) mixing, (c) wetting of the detection unit and (d) introduction into the detection unit using a force in a predetermined direction, and (a) weighing is performed in the predetermined range. It is preferable to use a force in a direction different from the direction, or (aa) perform centrifugation using a force in a predetermined direction. Thereby, the process of wetting a detection part and another process can be performed appropriately at an appropriate time.

Further, (a) weighing and (bb) wetting of the detection unit with the reagent are performed simultaneously, or (b) mixing and (bb) wetting of the detection unit with the reagent are performed simultaneously. Thereby, it is not necessary to perform a separate process to wet the detection unit, and the detection unit can be wetted more easily and quickly.
When the angle θ between the predetermined direction and the different direction is 0 <θ <180 °, or the microchip is configured by connecting the measuring unit, the mixing tank, the micro flow channel, and the detecting unit in series in this order. In this case, the microchip to be used can be easily designed, and a series of processes can be performed smoothly.
In addition, when the microchip is configured by arranging at least one reagent holding part upstream of the mixing tank or in the mixing tank or downstream of the detection part, the reagent and the sample are easily and reliably mixed. However, it is possible to perform the wetting process of the detection unit in a timely manner, which is advantageous.

Further, in the method of using the microchip of the present invention, (1) the microchip moves to the mixing vessel by the force in a predetermined direction, and the force in a direction different from the predetermined direction moves to the mixing tank. Or (2) a microchip, wherein the measuring section, the mixing tank, the microchannel, and the detection section are arranged so that the sample and / or reagent moves from the mixing tank to the detection section through the microchannel. However, it has a second mixing tank, and the measured sample and / or reagent moves to the mixing tank by a force in a direction different from the predetermined direction to form a mixed solution, and the force in the predetermined direction The metering unit, the mixing tank, so that the mixed liquid moves to the second mixing tank, and further, the mixed liquid moves from the second mixing tank to the detection unit through the microchannel by a force in a direction different from the predetermined direction. , 2nd mixing tank, micro flow path and Or detection portion is constituted is arranged, (3) the microchip is further coupled to both the mixing tank and the metering unit has a separating section you centrifugation the sample, and the predetermined direction or by different directions of the force, but the sample is moved to the metering unit from the separation unit, so as not to move in the mixing tank, the separation unit, the metering unit and the mixing vessel is constructed are arranged, (4) micro The chip moves the sample and / or reagent weighed to the mixing tank by a force in a predetermined direction, and the sample and / or reagent is detected from the mixing tank through the microchannel by a force in a direction different from the predetermined direction. The metering unit, the mixing tank, the microchannel, and the detection unit are arranged and configured to move to the unit, so that the mixing tank is configured to be arranged in a predetermined direction with respect to the metering unit. 5) The detection unit is arranged in a predetermined direction with respect to the mixing tank. Is, the detection unit is disposed in a predetermined direction relative to the mixing tank, microchannel connected to the detection section, and a mixing tank and a detection unit coupled to reach the detector from the mixing tank of the microchannel orientation However, in the case where the entire length of the microchannel is arranged and configured to be greater than −90 ° and less than + 90 ° with respect to a predetermined direction , the above-described effects can be more reliably realized. Can do.

In particular, the detection unit is arranged in a direction different from the predetermined direction with respect to the mixing tank, and the microchannel connected to the detection unit connects the mixing tank and the detection unit and detects from the mixing tank of the microchannel. When the direction to reach the portion is arranged so that it is greater than −90 ° and less than + 90 ° with respect to a direction different from the predetermined direction in the entire length of the microchannel, it is one direction on the microchip. The entire detection unit can be reliably wetted by a simple method of loading the force.
The microchannel of the present invention is a microchannel that is formed on a microchip and connects the mixing tank and the detection unit, and the direction from the mixing tank to the detection unit of the microchannel is the microchannel. in the total length of, for a given direction, greater than -90 °, + 90 formed in ° below become shape Ri Na, prior to detecting by the detecting unit and (c) samples, detection by the detection unit The detection unit is wetted with a mixed solution of a target reagent and a sample .

  By providing such a microchannel, a liquid substance can be smoothly moved from the mixing tank to the detection unit by using a simple method of applying a predetermined unidirectional force to the microchannel. It is possible to wet the detection unit in a dry state, and the contact angle of the liquid substance to the inner surface of the detection unit can be reduced. As a result, when a liquid substance is detected by the detection unit, it is possible to prevent detection interference due to generation of bubbles or adhesion of solid components to the inner surface of the detection unit, and accurate measurement can be performed. Furthermore, by loading forces in different directions, it is more advantageous that the sample and / or reagent detection section can be easily reciprocated.

The microchip of the present invention is a microchip including at least a mixing tank formed on a substrate, a detection unit, and a microchannel connecting the mixing tank and the detection unit, and the detection unit is predetermined with respect to the mixing tank. And the direction from the mixing tank of the microchannel to the detection unit is larger than −90 ° and less than + 90 ° with respect to the predetermined direction over the entire length of the microchannel. Ri Na is characterized by wetting said detector by a mixture of the reagent and the sample is (c) prior to detecting the sample by the detecting unit, detected in the detection unit.
Thereby, as described above, the entire detection unit can be reliably wetted by a simple method of applying a force in one direction to the microchip, and further, by applying a force in a different direction, the sample and / or Or it is more advantageous by being able to perform the reciprocation in the detection part of a reagent simply.

In particular, the microchip (1) further has a weighing unit,
When a force in a predetermined direction is applied, the weighed sample and / or reagent moves to the mixing tank,
When the force in a direction different from the predetermined direction is applied, the measuring unit, the mixing tank, the microchannel, and the detection unit are configured so that the sample and / or the reagent move from the mixing tank to the detection unit through the microchannel. (2) Furthermore, it has a measuring section and a second mixing tank,
When a force in a predetermined direction is applied, the weighed sample and / or reagent moves to the mixing tank,
When a force in a direction different from the predetermined direction is loaded, the moved sample and / or reagent moves to the second mixing tank, and when a force in the predetermined direction is loaded, the sample and / or reagent is moved to the second mixing tank. The metering unit, the mixing tank, the second mixing tank, the microchannel and the detection unit are arranged so as to move to the detection unit through the microchannel, or (3) the mixing tank is further metered Or the detector is arranged in a direction different from the predetermined direction with respect to the mixing tank, or (4) the mixing tank is in a predetermined direction with respect to the measuring unit. (5) The second mixing tank is arranged in a direction different from a predetermined direction with respect to the mixing tank, and the detection unit is arranged in a predetermined direction with respect to the second mixing tank. There are few chips in the mixing tank or in the mixing tank or downstream of the detection unit. (6) The microchip further has a separation unit, and the separation unit is loaded with a force in a direction different from the predetermined direction. In addition, in the case where the sample is connected to both the weighing unit and the mixing vessel so that the sample moves from the separation unit to the weighing unit but not to the mixing vessel, the above-described microchip usage method is used. Can be reliably applied, and more accurate and quick sample detection can be realized.

  According to the present invention, by performing an appropriate treatment such as hydrophilization over the entire surface of the microchannel regardless of the size of the microchannel, even if the amount of sample is small, sample adhesion and bubbles Can be prevented, and as a result, the flow of the sample to be measured is not disturbed, and the microchip can perform accurate analysis by introducing the sample appropriately into the measurement system. It is an object of the present invention to provide a method for using the above, a microchannel and a microchip.

(1) Micro-channel The micro-channel of the present invention connects a mixing tank formed on a microchip and a detection unit, and can pass a sample and / or a reagent (preferably a liquid). As described above, the microchip is formed in a desired shape two-dimensionally and / or three-dimensionally. In the microchannel, the direction from the mixing tank to the detection unit is greater than −90 ° and less than + 90 ° with respect to a predetermined direction (that is, a predetermined direction) over the entire length of the microchannel. (Within a range of ± 90 °, where −90 ° and + 90 ° are excluded, hereinafter the same meaning). In other words, when a force in a predetermined direction, for example, a centrifugal force in one direction, is applied, almost all of the liquid substance contained in the mixing tank is surely passed through the microchannel to the detection unit. It is formed in a shape that can be moved.

Specifically, in the microchip 10 as shown in FIG. 1A, when the micro flow path 4 that connects the mixing tank 2 and the detection unit 3 is formed in one linear shape, Usually, the straight line is constituted by the shortest distance connecting the mixing tank 2 and the detection unit 3. Therefore, if the predetermined direction is the direction indicated by G1 in FIG. 1 (a), the shortest distance described above and the direction from the mixing tank 2 toward the detection unit 3 is ± 90 with respect to G1. It means within the range of °.
Further, when formed with two or more straight lines, for example, as shown in FIG. 1B, the direction (b1 and b2) from the mixing tank 2 of the microchannel 4 to the detection unit 3 is This means that the entire length of the microchannel 4 is formed within a range of ± 90 ° with respect to the predetermined direction G1.

  Furthermore, as shown in FIG. 1C, when the micro flow path 4 connecting the mixing tank 2 and the detection unit 3 is formed by a curve, the micro flow from the mixing tank 2 to the detection unit 3 The direction of the path 4 (c1 to c4), that is, the tangent at an arbitrary point of the microchannel 4 from the mixing tank 2 to the detection unit 3 is ± 90 with respect to the predetermined direction G1 over the entire length of the microchannel 4. It means that it is formed within the range of °.

In a microchannel having such a shape, preferably a microchip including the microchannel, a liquid substance (sample and / or reagent) is loaded with a force in only one predetermined direction, thereby mixing tank The liquid substance can be smoothly moved from the sensor to the detection unit.
In addition, the liquid substance which moved to the detection part can be returned to a mixing tank by applying the force to the arbitrary directions different from the force with respect to a predetermined direction. Thereby, it becomes possible to reciprocate the liquid substance between the mixing tank and the detection unit by selecting two predetermined directions and sequentially applying a force in these directions. The arbitrary direction here includes a direction in which the direction from the detection unit to the mixing tank is within a range of ± 90 ° with respect to the arbitrary direction over the entire length of the microchannel.

The cross section of the microchannel (cross section perpendicular to the fluid flow direction) is, for example, a polygon such as a quadrangle or a trapezoid, and a shape with rounded corners, a circle, an ellipse, a dome, or a left and right Any shape such as an asymmetrical non-uniform shape may be used. The microchannel is for flowing a small amount of liquid sample, and its size and length are not particularly limited as long as the purpose is appropriately achieved. For example, the cross-sectional area is 0.0001 to ² mm ^2. Examples include those having a length of about 10 to 100 mm. In particular, the smaller the cross-sectional area of the channel, the greater the flow of fluid in the micro-channel, the influence of bubbles generated in the fluid, the adhesion of solids, and the like. The present invention is advantageous with respect to those of about 0001 to ¹ mm ² . The microchannels may have the same cross-sectional shape and size over the entire length, but may have partially different shapes and sizes.

  The microchannel may be, for example, PET (polyethylene terephthalate, PDMS (polydimethylsiloxane), PMMA (polymethylmethacrylate), PC (polycarbonate), PP (polypropylene), PS (polystyrene), depending on the method of producing them. It is formed using any of organic compounds such as PVC (polyvinyl chloride), polysiloxane, allyl ester resin, cycloolefin polymer, Si rubber, or inorganic compounds such as Si, Si oxide film, quartz, glass, and ceramic. These materials may be transparent, or may be translucent, opaque, or light-shielding by incorporating a pigment such as carbon black.

  The microchannel can be easily produced by using a method known in the art. For example, a mold having a shape corresponding to a desired microchip and microchannel is prepared. This mold can be formed by mechanical processing. Next, PET is molded into this mold to obtain a substrate on which a pattern corresponding to the microchannel is transferred. Finally, by attaching two substrates to each other so that the patterns face each other, a microchip having a microchannel at a desired position can be formed. Only one substrate having a pattern corresponding to the microchannel may be used, and the other may be a flat substrate. Further, instead of molding using a mold, an injection molding method or an imprint method may be used. Furthermore, one or both of the flat substrates may be directly subjected to a photolithography process, mechanical processing, or the like to obtain a substrate on which a pattern corresponding to the microchannel is transferred.

(2) Microchip configuration For example, a microchip is mainly formed by bonding a first substrate and a second substrate having patterns of various shapes with concave portions on one or both sides, for example, by thermocompression bonding, an adhesive, or the like. Configured. Note that these substrates can be formed according to the manufacturing method described above.

  Such microchips analyze and detect various biological materials (samples, mainly in a liquid state) such as DNA, enzymes, proteins, viruses and cells in various fields such as medicine, food, and drug discovery. It is used as a substrate for reaction, measurement, etc., for example, clinical analysis chip, environmental analysis chip, gene analysis chip (DNA chip), protein analysis chip (proteome chip), sugar chain chip, chromatographic chip, It includes all chips provided under various names such as cell analysis chips, pharmaceutical screening chips and the like.

Such a microchip includes at least a mixing tank, a detection unit, and a microchannel that connects the mixing tank and the detection unit as described above. , Discharge ports and / or waste liquid reservoirs, centrifuges, metering units, mixing tanks, reagent holding units, mixers, etc., each one or more, themselves or various two-dimensional and / or three-dimensional connections It is arranged with a flow path having a linear or bent or curved pattern.
In particular, it is preferable that the measuring unit, the mixing tank, the microchannel, and the detection unit are configured to be connected in series in this order.

The mixing tank is usually a space for mixing a sample and a reagent, and the size thereof is not particularly limited, and has a size sufficient to realize this purpose. However, it can be appropriately set according to the type and amount of these substances. For example, about 10 ^<-2 > -10 ^{< 3} > mm ^{< 3} > is mentioned. In addition, the shape of the mixing tank is not particularly limited as long as it is suitable for the purpose of mixing. For example, both the plane and the cross-sectional shape are polygons such as a quadrangle and a trapezoid, and the corners are rounded. Any shape such as a circular shape, an oval shape, a dome shape, or an asymmetrical non-uniform shape may be used.
In addition, the mixing tank may have a function as the reagent holding unit described above by holding not only the mixing but also the reagent therein, and a plurality of mixing tanks may be formed.

The detection unit is a space for detecting a sample, and its size and shape are not particularly limited. For example, the detection unit is appropriately set according to the detection method (method), the type and amount of the sample and / or reagent, and the like. can do. A specific example is approximately 1 to 10 ³ mm ^3. Moreover, the shape can be set to various shapes like a mixing tank.
In particular, the detection unit is preferably detected (analyzed and measured) using an optical method. Therefore, in the detection unit, a shape capable of ensuring a predetermined length of an optical path capable of entering and exiting light so that the sample can be irradiated with light and the passage, reflection, and scattered light of the light can be detected. The size is preferred. For example, a detector having a certain shape with a length of about 1 to 50 mm and a cross-sectional area of about 0.1 to 100 mm ² is used.

The measuring unit has a function of measuring the amount, usually the volume, of a sample to be used for measurement (detection), and extracting only a predetermined amount. It can be adjusted by narrowing or expanding the space to be constructed. The measurement by the measurement unit is formed in a shape that can be executed by generating a force (for example, centrifugal force) in a predetermined direction in the microchip.
Sample introduction port, discharge port, waste liquid reservoir, separation unit, reagent holding unit, mixer, etc. are appropriately selected according to the type, amount, analysis method, etc. of the sample from those that can ensure the shape and function known in the field Or it can change and use.

  In the microchip of the present invention, the detection unit is arranged in a predetermined direction with respect to the mixing tank. Here, “arranged in a predetermined direction with respect to the mixing tank” means that the detection unit (even if all or part of it is within a range of ± 90 ° with respect to a straight line extending in an arbitrary direction from the mixing tank. Means good). In other words, when a force directed in an arbitrary direction with respect to the mixing tank, for example, a centrifugal force, is applied, all of the liquid substance contained in the mixing tank passes through the above-described microchannel and the detection unit. It is arranged to move to.

  For example, as shown in FIG. 2 (c), when the straight line going in an arbitrary direction is the direction of g1, the detection unit 3 is arranged in the g1 direction with respect to the mixing tank 2 or within ± 90 ° thereof. As shown in FIG. 4 (d), when the straight line going in an arbitrary direction is the direction of g2, the detection unit 3 is arranged in the g2 direction or within ± 90 ° of the mixing tank 2a. .

  The measuring unit is set so that the mixing tank is arranged in a predetermined direction with respect to the measuring unit. Here, “arranged in a predetermined direction” has the same meaning as described above. For example, when the “predetermined direction” is the g2 direction as shown in FIG. 2B, the mixing tank 2 is arranged in the g2 direction of the measuring unit 8. However, in this case, it is preferable that the detection unit 3 described above is arranged in a direction (for example, g1) different from the predetermined direction in this case. In other words, when a force in the second direction (for example, g2) is loaded, a weighed sample or the like moves to the mixing tank 2, and when a force in the first direction (for example, g1) is loaded. In addition, it is preferable that the measuring unit 8, the mixing tank 2, the microchannel 4, and the detection unit 3 are arranged so that the sample or the like moves from the mixing tank 2 through the microchannel 4 to the detection unit 3.

  For example, as shown in FIG. 4B, when two mixing tanks 2, 2 a are arranged, the weighing unit 8 is configured so that the first mixing tank 2 is in a predetermined direction with respect to the weighing unit 8. (For example, g2 direction shown in FIG.4 (b)), and the 2nd mixing tank 2a is arrange | positioned with respect to the 1st mixing tank 2 in the direction (for example, g1) different from a predetermined direction. It is preferable. In other words, when a force in the second direction (for example, g2) is loaded, the weighed sample and / or reagent moves to the mixing tank 2, and a force in the first direction (for example, g1) is loaded. When the sample and / or reagent moved to the second mixing tank 2a, and when a force in the second direction is applied, the sample and / or reagent passes through the microchannel 4 from the second mixing tank 2a. The measuring unit 8, the mixing tank 2, the second mixing tank 2 a, the micro flow path 4, and the detecting unit 3 are preferably arranged so as to move to the detecting unit 3.

  The separation unit is a part intended to separate, preferably centrifuge, the sample and separate the sample into two or more according to the difference in specific gravity, and the weighing unit has a first direction (for example, g1) with respect to the separation unit. It is set to be placed on the side. In other words, the sample is arranged so as to move from the separation unit to the measurement unit when a force is applied in the first direction. However, at this time, the separation unit is preferably configured to be connected to both the weighing unit and the mixing vessel so that the sample does not move directly from the separation unit to the mixing vessel.

  It is preferable that one or two or more reagent holding units are arranged upstream of the mixing tank, in the mixing tank, or downstream of the detection unit. Here, “upstream” is closer to the initial stage in a series of flows in which the sample is finally detected (analyzed, measured) by introducing the sample into the microchip and mixing with a reagent or the like. (That is, the one closer to the sample inlet).

(3) How to use the microchip Normally, the microchip is used in a disposable manner and with minimal steps in order to analyze a small amount of sample quickly and accurately in a shorter time. However, in the present invention, in order to prevent the solid components such as proteins and oils and fats in the sample in the detection unit from adhering to the inner surface, the generation of bubbles, etc., the sample, reagent introduced into the microchip or these Before detecting (preferably immediately before) the liquid mixture in the detection unit, the detection unit is once passed through the detection unit to wet the inner surface of the detection unit, and in such a wet state (before complete drying), detection is performed again. This is a method of using a microchip in which a test sample is introduced into a part and detected.

As the microchip that can be used in the method of use of the present invention, those described above can be preferably used.
The means for passing the sample and / or reagent in advance through the detection unit is not particularly limited, and uses, for example, suction or discharge by a pump; centrifugal force; force in at least one direction due to dropping, vibration or shaking; be able to. Among these, it is preferable to use centrifugal force.
In the present invention, the phrases “... loading a force in the direction” and “... using the force in the direction” move the liquid present in the microchip in the direction. It means giving force to this liquid. Further, “using a centrifugal force” can be rephrased as “rotating the microchip around a predetermined rotation axis set inside or outside the microchip”.

  In the present invention, a liquid substance is passed through the detection unit before detection, but this liquid substance may be a sample, may be a reagent alone, or may be a sample and a reagent. It may be a mixture. In addition, passing through the detection unit may be a one-time outbound path or a return path, or may be performed twice or more so as to reciprocate. It should be noted that it is appropriate to perform the passage of the liquid substance to the detection unit once only in one step. That is, wetting of the liquid substance over the entire length of the detection part can be realized by one step of applying a force in one direction. Similarly, the sample can be separated, weighed, mixed, etc. by one step of applying a force in one direction.

Specific examples of methods of using the microchip optionally include (aa) centrifuging the introduced sample and / or (bb) passing the reagent through the detection section and wetting the detection section, At least (a) a step of weighing a sample, (b) a step of mixing the weighed sample with a reagent, (c) a step of wetting a liquid mixture of the sample with the reagent through the detection unit and wetting the detection unit, (d) This includes the step of detecting the sample by introducing the liquid mixture into the already wet detection unit. Among these steps, (b) mixing is performed using force in a predetermined direction, and (a) weighing and (c) detection. It is preferable that the wetting of the part and (d) introduction into the detection part is performed using a force in a direction different from the predetermined direction.
Alternatively, (b) mixing, (c) wetting of the detection unit, and (d) introduction into the detection unit are performed using a force in a predetermined direction, and (a) a force in a direction different from the predetermined direction is used. You may use it.

In any of these cases, when performing centrifugation, it is preferable to perform (aa) centrifugation using a force in a predetermined direction.
In each step described above, (a) measurement may be performed at the same time as (bb) wetting of the detection unit, or (b) mixing may be performed at the same time as (bb) wetting of the detection unit. .
The angle θ between the predetermined direction and the different direction can be 0 <θ <180 °, and preferably about 60 ≦ θ ≦ 120 °.

Embodiment 1
In this embodiment, a method of using the microchip 10 shown in FIG. 1A will be described.
First, the microchip 10 is prepared. The microchip 10 is configured by connecting a sample introduction port 6, a channel 9, a mixing tank 2, a microchannel 4, and a detection unit 3 in series. The detection unit 3 is arranged, for example, on the arrow direction side indicated by G1 with respect to the mixing tank 2, and the micro flow path 4 connecting the mixing tank 2 and the detection unit 3 is In the entire length, it intersects with the g1 direction within a range of ± 90 °, that is, in the direction (arrow X) from the mixing tank 2 toward the detection unit 3, the direction of the angle of θ ° (for example, in the G1 direction) + 50 ° direction).

Next, a sample and a reagent containing a test substance are sequentially introduced into the microchip 10 from the sample introduction port 6 and passed through the flow path 9.
Thereafter, by applying a centrifugal force (arrow G2) in a direction different from the above-described predetermined direction, for example, a direction of −90 ° with respect to the predetermined direction, that is, a predetermined rotation axis (for example, FIG. 1A). By rotating around the point B), the sample and the reagent pass through the flow path 9 and are introduced into the mixing tank 2 to form a uniform mixed solution 5a.

Next, the liquid mixture 5a passes through the microchannel 4 by the centrifugal force in the G1 direction, enters the detection unit 3, and wets the entire inner surface of the detection unit 3 with the liquid mixture 5b.
Subsequently, the mixed solution 5b returns to the mixing tank 2 again by the centrifugal force in the direction of G2.
Finally, the mixed solution 5a is again introduced into the detection unit by the centrifugal force in the G1 direction, and detection can be performed.

  By using such a microchip, the sample can be freely moved and reciprocated in the microchip by a simple method of applying a centrifugal force or rotating the microchip. In addition, since the detection unit is preliminarily wetted with the mixed solution of the sample and the reagent instead of being in a dry state, the wettability of the test substance (mixed solution) is increased and the contact angle is decreased when the mixed solution is introduced at the time of detection. be able to. As a result, about 30 bubbles that can be visually recognized were generated when the entire inner surface of the detection unit 3 was wetted with the first mixed solution 5b. However, when the mixed solution 5b was introduced into the detection unit 3 for the second time or later, air The generation of pockets could be prevented, and no bubbles were generated during detection, and further, accurate measurement could be performed without causing solid components to adhere.

  In the above method, after the entire inner surface of the detection unit 3 is wetted with the mixed solution by the centrifugal force in the G1 direction, the centrifugal force in the G2 and G1 directions may be loaded for two cycles or more. Thereby, mixing of a sample and a reagent can be performed more uniformly.

Embodiment 2
First, the microchip 20 shown in FIG. The microchip 20 connects the sample introduction port 6, the separation unit 7, the measurement unit 8, the reagent holding unit 11, the second reagent holding unit 12, the channel 9, the mixing tank 2, the microchannel 4, and the detection unit 3. Configured.
The detection unit 3 is arranged, for example, on the side of the mixing tank 2 in a first direction (for example, an arrow direction represented by g1) intended to load a centrifugal force, and further, the mixing tank 2 and the detection unit. 3 in the entire length of the microchannel 4 so as to intersect the g1 direction within a range of ± 90 ° with respect to the g1 direction, that is, from the mixing tank 2 to the detection unit 3. For example, it is arranged so as to face in the direction of about + 60 ° in the g1 direction.

The mixing tank 2 is disposed on the side of the arrow indicated by g2 (that is, −90 ° with respect to g1) with respect to the measuring unit 8.
The measuring unit 8 is arranged on the arrow direction side indicated by g1 with respect to the separating unit 7. A waste liquid reservoir 13 is disposed downstream of the measuring unit 8.

The separation unit 7 is arranged on the side of the arrow direction indicated by g2 with respect to the sample injection port 6. In addition, the separation unit 7 moves the separated sample from the separation unit 7 to the weighing unit 8 by centrifugal force in the g1 direction, but does not move to the mixing vessel 2. Are connected to both.
Note that the reagent holding unit 11 and the second reagent holding unit 12 hold different reagents 16a and second reagents 16b, respectively, and the flow path 9 (that is, the mixing tank) between the measuring unit 8 and the mixing tank 2 is retained. 2 upstream), respectively.

Next, as shown in Step 1 of FIG. 2A and FIG. 7, blood 14 is introduced into the microchip 20 as a sample containing the test substance from the sample introduction port 6.
Subsequently, as shown in step 2 of FIG. 2B and FIG. 7, the blood is centrifuged into blood cells 14 a and plasma 14 b in the separation unit 7 by the centrifugal force in the g2 direction.
Thereafter, as shown in Step 3 of FIG. 2C and FIG. 7, the separated plasma 14b is introduced into the measuring unit 8 by the centrifugal force in the g1 direction. At this time, in the measuring unit 8, a predetermined amount of plasma 14b is held in the measuring unit 8, and excess plasma 14b is collected in the waste liquid reservoir 13, and as a result, a predetermined amount of plasma 14b is measured.

Next, as shown in Step 4 of FIG. 2 (d) and FIG. 7, a predetermined amount of plasma 14b that has been measured is introduced into the mixing tank 2 by the centrifugal force in the g2 direction. At the same time, the reagent 16a and the second reagent 16b held in the reagent holding unit 11 and the second reagent holding unit 12 are also introduced into the mixing tank 2, respectively, and the plasma 14b, the reagent 16, and the second reagent 16b Mixture 15 is provided.
Thereafter, as shown in Step 5 of FIG. 2 (e) and FIG. 7, the mixed liquid 15 passes through the micro flow path 4 by the centrifugal force in the g <b> 1 direction and enters the detection unit 3. Is wetted with the mixture 15.

Subsequently, as shown in Step 6 of FIG. 2 (f) and FIG. 7, the mixed solution 15 is returned to the mixing tank 2 by the centrifugal force in the direction of g 2 again.
Finally, as shown in step 7 of FIG. 2 (g) and FIG. 7, the mixed liquid 15 is introduced into the detection unit by the centrifugal force in the direction of g1 again. Thereafter, the sample is detected by irradiating the test substance with light L and detecting the transmitted light with a light receiving element such as a photodiode.

By using such a microchip, as in Embodiment 1, the wettability of the detection unit can be increased, the contact angle can be reduced, the generation of bubbles during detection and the adhesion of solid components can be prevented, Accurate measurement can be performed.
Also in the above method, as in the first embodiment, the detection unit is wetted by the centrifugal force in the g1 direction in step 5 of FIGS. 2 (e) and 7, and step 6 in FIGS. 2 (f) and 7 is performed again. The mixture 15 may be returned by centrifugal force in the direction of g2 of two cycles or more. Thereby, mixing of a sample and a reagent can be performed more uniformly.

Embodiment 3
First, the microchip 30 shown in FIG. The microchip 1 has substantially the same configuration as the microchip 20 in FIG. 2A except that the shapes of the reagent holding unit 11 and the second reagent holding unit 12 and the connection position upstream of the mixing tank 2 are slightly different. is there.
Next, as shown in FIGS. 3 (a) and 3 (b) and steps 1 and 2 in FIG. 8, blood 14 is introduced into the microchip 20, and centrifugal force is applied in the g2 direction. Plasma 14b is separated. At this time, the respective reagents 16 a and 16 b held in the reagent holding unit 11 and the second reagent holding unit 12 move to the mixing tank 2 and are mixed to give the mixed reagent 16.

Thereafter, as shown in Step 3 of FIG. 3 (c) and FIG. 8, the plasma 14b is loaded with a centrifugal force in the direction g1 and introduced into the measuring unit 8 and weighed. At the same time, the mixed reagent 16 in the mixing tank 2 passes through the microchannel 4 and enters the detection unit 3, so that the entire inner surface of the detection unit 3 is wetted with the mixed reagent 16.
Next, as shown in steps 4 and 5 of FIG. 3 (d) and FIG. 8, a predetermined amount of the measured plasma 14b is introduced into the mixing tank 2 by applying a centrifugal force in the direction g2. At the same time, the mixed reagent 16 is returned to the mixing tank 2, and a mixed solution 15 of the plasma 14b and the mixed reagent 16 is given.

Thereafter, as shown in FIGS. 3E to 3G and steps 6 to 8 of FIG. 8, the detection unit is wetted and returned in the same manner as in the second embodiment to detect the sample.
The same effect as that of the first embodiment can be obtained by using such a microchip.

Embodiment 4
First, the microchip 40 shown in FIG. 4A is prepared. The microchip 40 includes a sample introduction port 6, a separation unit 7, a measurement unit 8, a reagent holding unit 11, a channel 9, a mixing tank 2, a second mixing tank 2 a, a microchannel 4, and a detection unit 3. It is configured.
The detection unit 3 is arranged in a predetermined direction intended to load a centrifugal force with respect to the second mixing tank 2a, for example, on the arrow direction side indicated by g2, and further, the second mixing tank 2a and the detection unit 3 in the entire length of the microchannel 4 so as to intersect within the range of ± 90 ° with respect to the g2 direction, that is, the direction from the second mixing tank 2a to the detection unit 3 For example, it is arranged so as to be directed in the direction of −40 ° in the g2 direction.

The 2nd mixing tank 2a is arrange | positioned with respect to the mixing tank 2 in the arrow direction (namely, +90 degrees with respect to g2) represented by g1. Moreover, the mixing tank 2 is arrange | positioned with respect to the measurement part 8 in the arrow direction side represented by g2.
The measuring unit 8 is arranged on the arrow direction side indicated by g1 with respect to the separating unit 7. A waste liquid reservoir 13 is disposed downstream of the measuring unit 8.

The separation unit 7 is arranged on the side of the arrow direction indicated by g2 with respect to the sample injection port 6. In addition, when the centrifugal force in the g1 direction is loaded, the separation unit 7 moves from the separation unit 7 to the measurement unit 8 but does not move to the mixing tank 2 so that the separation unit 7 moves to the mixing tank 2. It is connected to both the mixing tank 2.
Different reagent 16a and second reagent 16b are held in the reagent holding part 11 and the second mixing tank 2a, respectively, and the sample holding part 11 has a flow path 9 (that is, a flow path 9 between the measuring part 8 and the mixing tank 2 (that is, , Upstream of the mixing tank).

Next, as shown in step 1 of FIG. 4A and FIG. 9, blood 14 is introduced into the microchip 40.
Subsequently, as shown in Step 2 of FIG. 4B and FIG. 9, a centrifugal force is applied in the g2 direction, and the blood is centrifuged into blood cells 14a and plasma 14b in the separation unit 7. At the same time, the second reagent 16b held in the second mixing tank 2a passes through the microchannel 4, enters the detection unit 3, and wets the entire inner surface of the detection unit 3 with the second reagent 16b.

Thereafter, as shown in Step 3 of FIG. 4C and FIG. 9, the separated plasma 14 b is introduced into the measuring unit 8 with a centrifugal force applied in the g1 direction. At the same time, the second reagent 16b returns from the detection unit 3 to the second mixing tank 2a.
Next, as shown in step 4 of FIG. 4 (d) and FIG. 9, a predetermined amount of the measured plasma 14b is introduced into the mixing tank 2 by applying a centrifugal force in the direction g2, and a reagent holding unit. The reagent 16 held in 11 is also introduced into the mixing tank 2, and the first mixing is performed to give the first mixed solution 15a of the plasma 14b and the reagent 16a. At the same time, the second reagent 16b moved to the second mixing tank 2a passes through the microchannel 9 again to the detection unit 3 and wets the inner surface of the detection unit 3.

Thereafter, as shown in FIG. 4 (e) and steps 5 and 6 of FIG. 9, by applying a centrifugal force in the g1 direction, the second reagent 16b returns from the detection unit 3 to the second mixing tank 2a. At the same time, the first mixed liquid 15a moves from the mixing tank 2 to the second mixing tank 2a, and the second mixing is performed to give the mixed liquid 15.
Thereafter, as shown in Step 7 of FIG. 4 (f) and FIG. 9, by applying a centrifugal force in the g2 direction, the mixed solution 15 passes through the microchannel 4 and enters the detection unit 3, and then the detection unit 3. Wet the entire inner surface of the mixture with the mixture 15.

Subsequently, as shown in Step 8 of FIG. 4 (g) and FIG. 9, the mixed liquid 15 is returned to the second mixing tank 2a by applying a centrifugal force again in the direction of g1.
Finally, as shown in step 9 of FIG. 4 (h) and FIG. 9, by applying a centrifugal force again in the direction of g2, the liquid mixture 15 is introduced into the detection unit 3 to perform detection.
By using such a microchip, as in Embodiment 1, the wettability of the detection unit can be increased, the contact angle can be reduced, the generation of bubbles during detection and the adhesion of solid components can be prevented, Accurate measurement can be performed.

Embodiment 5
First, a microchip 50 shown in FIG. The microchip 50 is configured by connecting a sample introduction port 6, a separation unit 7, a measurement unit 8, a flow channel 9, a mixing tank 2, a microchannel 4, a detection unit 3, and a reagent holding unit 11. Except for the position of the holding unit 11 being different, the arrangement relationship of the sample introduction port 6, the separation unit 7, the measurement unit 8, the flow channel 9, the mixing tank 2, the micro flow channel 4 and the detection unit 3 is substantially the same as that of the second embodiment. It is the same.

The reagent holding unit 11 is disposed downstream of the detection unit 3 and the detection unit 3 is disposed on the reagent holding unit 11 on the arrow direction side indicated by g2, for example.
Further, the reagent holding unit 11 and the mixing tank 2 hold different reagents 16a and second reagents 16b, respectively.

Next, as shown in Step 1 of FIG. 5A and FIG. 10, blood 14 is introduced into the microchip 20.
Subsequently, as shown in Step 2 of FIG. 5B and FIG. 10, a centrifugal force is applied in the g2 direction, and the blood is centrifuged into blood cells 14a and plasma 14b in the separation unit 7. At the same time, the second reagent 16b held in the reagent holding unit 11 is introduced into the detection unit 3, wets the detection unit 3, and is further introduced into the mixing tank 2 through the microchannel 4, and the reagent 16a and Quickly, mixed reagent 16 is provided.

Thereafter, as shown in Step 3 of FIG. 5C and FIG. 10, the separated plasma 14 b is introduced into the measuring unit 8 with a centrifugal force applied in the g1 direction. At the same time, the reagent 16 introduced into the mixing tank 2 is introduced into the detection unit 3 through the microchannel 4 and wets the detection unit 3.
Next, as shown in Step 4 of FIG. 5 (d) and FIG. 10, a predetermined amount of the measured plasma 14b is introduced into the mixing tank 2 by applying a centrifugal force in the direction g2, and the detection unit 3 The mixed reagent 16 having been wetted is introduced into the mixing tank 2, and a mixed solution 15 of the plasma 14b, the reagent 16, and the second reagent 16b is provided.

Thereafter, as shown in FIGS. 5E to 5G and steps 5 to 7 in FIG. 10, detection is performed in the same manner as in the second embodiment.
By using such a microchip, as in the second embodiment, the wettability of the detection unit can be increased, the contact angle can be reduced, the generation of bubbles during detection and the adhesion of solid components can be prevented, Accurate measurement can be performed.

Embodiment 6
First, a microchip 60 shown in FIG. 6A is prepared. The microchip 60 is configured by connecting the sample introduction port 6, the separation unit 7, the reagent holding unit 11, the second reagent holding unit 12, the measuring unit 8, the flow path 9, the mixing tank 2, the mixer 17, and the detection unit 3. The positional relationship among the sample introduction port 6, the separation unit 7, the measurement unit 8, the reagent holding unit 11, the second reagent holding unit 12, the flow path 9, and the mixing tank 2 is substantially the same as in the second embodiment. is there.

The detection unit 3 is arranged in a predetermined direction with respect to the mixing tank 2, for example, a first direction intended to load a centrifugal force, the arrow direction side indicated by g1, but this positional relationship is not necessary. Also good. The flow path 9 connecting the mixing tank 2 and the detection unit 3 has three H-type mixers 17 arranged in series in the middle.
Pump holes 18a and 18b for connecting a pump are formed upstream of the mixing tank 2 and downstream of the detection unit 3, respectively.

Next, as shown in step 1 of FIG. 6A and FIG. 11, blood 14 is introduced into the microchip 20.
Subsequently, as shown in Step 2 of FIG. 6B and FIG. 11, a centrifugal force is applied in the g2 direction, and the blood is centrifuged into blood cells 14a and plasma 14b in the separation unit 7. At the same time, the reagent 16 a and the second reagent 16 b held in the reagent holding unit 11 and the second reagent holding unit are introduced into the mixing tank 2 to give the mixed reagent 16.

  Thereafter, as shown in Step 3 of FIG. 6C and FIG. 10, the separated plasma 14 b is introduced into the measuring unit 8 with a centrifugal force applied in the g1 direction. At this time, the reagent 16 held in the mixing tank 2 is not introduced into the detection unit 3 only by the centrifugal force in the g1 direction due to the presence of the H-type mixer 17.

Next, as shown in Step 4 of FIG. 6 (d) and FIG. 10, a predetermined amount of the plasma 14b is introduced into the mixing tank 2 by applying a centrifugal force in the direction g2, and the mixed reagent 16 And a mixed solution 15 of plasma 14b, reagent 16a, and second reagent 16b is provided.
Thereafter, as shown in Step 5 of FIG. 6 (e) and FIG. 10, by sucking from the pump hole 18a, the mixed solution 15 is sufficiently mixed by the H-type mixer 17, and further passes through the flow path 9a, and the detection unit 3 and the entire inner surface of the detection unit 3 is wetted with the liquid mixture 15.

Subsequently, as shown in step 6 of FIG. 6 (f) and FIG. 10, the mixed solution 15 is returned to the mixing tank 2 by suction from the pump hole 18 b.
Finally, as shown in step 7 of FIG. 6 (g) and FIG. 10, by sucking again from the pump hole 18b, the liquid mixture 15 is introduced into the detection unit and detection is performed.

  By using such a microchip, as in the second embodiment, the wettability of the detection unit can be increased, the contact angle can be reduced, the generation of bubbles during detection and the adhesion of solid components can be prevented, Accurate measurement can be performed.

  The present invention relates to a clinical analysis chip, an environmental analysis chip, a gene analysis chip (DNA chip), a protein analysis chip (proteome chip), a sugar chain chip, a chromatographic chip, used in the fields of medicine, food, drug discovery, etc. It can be used for various substrates that can be applied to gases and liquids called cell analysis chips and pharmaceutical screening chips.

The top view which shows one Embodiment of the microchannel of this invention. It is a top view which shows a series of operation | movement for demonstrating the usage method of the microchip of this invention. It is a top view which shows a series of operation | movement for demonstrating the usage method of another microchip of this invention. It is a top view which shows a series of operation | movement for demonstrating the usage method of another microchip of this invention. It is a top view which shows a series of operation | movement for demonstrating the usage method of another microchip of this invention. It is a top view which shows a series of operation | movement for demonstrating the usage method of another microchip of this invention. It is a flowchart which shows a series of operation | movement of FIG. 4 is a flowchart showing a series of operations in FIG. 3. 5 is a flowchart showing a series of operations in FIG. 4. 6 is a flowchart showing a series of operations in FIG. 5. It is a flowchart which shows a series of operation | movement of FIG.

Explanation of symbols

10, 20, 30, 40, 50, 60 Microchip 2 Mixing tank 2a Second mixing tank 3 Detection section 4 Micro flow path 5a, 5b Mixed liquid of sample and reagent 6 Sample inlet 7 Centrifugal section 8 Measuring section 9 , 9a Channel 11 Sample holder 12 Second sample holder 13 Waste liquid reservoir 14 Blood 14a Blood cell 14b Plasma 15 Mixed liquid 16 Mixed reagent 16a Reagent 16b Second reagent 17 Mixer 18a, 18b Pump hole

Claims (24)

A reagent holding unit for holding a reagent formed on the substrate, a mixing vessel, a detection unit, and at least a microchannel having the same cross-sectional area over the entire length by connecting the mixing vessel and the detection unit ,
The mixing tank, the microchannel, and the detection unit are connected in series in this order.
That the microchip method uses the first direction and the first angle theta is 0 between the direction of <theta <second direction the microchip centrifugal force added sequentially to a is 180 ° There,
(B) introducing the sample and the reagent into the mixing tank by centrifugal force in the second direction, and mixing the sample with the reagent in the mixing tank;
(C) Before detecting the sample by the detection unit, the detection liquid is introduced into the detection unit by introducing a mixed solution of the reagent and the sample to be detected by the detection unit by the centrifugal force in the first direction. The process of wetting the part,
(C1) returning the mixed liquid to the mixing tank by centrifugal force in the second direction;
(D) a step of detecting a sample in the detection unit by introducing a liquid mixture by centrifugal force in the first direction into the detection unit that has been wetted by the wetting step of (c);
Only including,
Due to the centrifugal force in the first direction, the mixed solution moves from the mixing tank to the detection unit through the micro flow channel, and the mixed solution moves from the detection unit to the micro flow channel by the centrifugal force in the second direction. A method of using a microchip , wherein the mixing tank, the microchannel, and the detection unit are disposed at a position where the mixing tank, the microchannel, and the detection unit are moved through . The microchip further includes a weighing unit,
Further comprising the step of weighing the sample in (a) weighing unit,
The method according to claim 1, wherein in the step (b) , the sample weighed in the step (a) is mixed with a reagent. The microchip further includes a sample inlet and a separation unit,
The method according to claim 2, further comprising (aa) centrifuging the introduced sample in the separation unit.   The method according to claim 2 or 3, further comprising the step of (bb) passing the reagent through the detection unit and wetting the detection unit before the step of (c) wetting. The method according to any one of claims 2 to 4 , wherein (a) weighing is performed by centrifugal force in the first direction . (Aa) The method according to any one of claims 3 to 5 , wherein the centrifugation is performed by a centrifugal force in the second direction . (A) weighing and process according to any one of claims 4-6 for performing simultaneously the wetting of the detection unit according to (bb) reagent. The method according to any one of claims 2 to 7 , wherein the microchip is configured by connecting a measuring section, a mixing tank, a microchannel, and a detection section in series in this order. The method according to any one of claims 1 to 8 , wherein the microchip is configured by arranging at least one of the reagent holding parts upstream of the mixing tank or in the mixing tank or downstream of the detection part. The separation unit is connected to both the mixing tank and the metering unit, by the centrifugal force in the first direction, so that although the sample is moved to the metering unit from the separation unit, does not move in the mixing vessel The method according to claim 3 , wherein the separation unit, the measurement unit, and the mixing tank are arranged. The method according to claim 2 , wherein the mixing tank is arranged in the second direction with respect to the measuring unit. The detection unit is arranged in the first direction with respect to the mixing tank, and the micro flow path connected to the detection unit connects the mixing tank and the detection unit, and the direction from the mixing tank of the micro flow path to the detection unit The method according to any one of claims 1 to 11 , wherein the microchannel is disposed so that the total length of the microchannel is greater than -90 ° and less than + 90 ° with respect to the first direction . A reagent holding unit, a mixing tank, a second mixing tank, a detection unit, and a micro that has the same cross-sectional area over the entire length by connecting the second mixing tank and the detection unit to hold the reagent formed on the substrate Comprising at least a flow path,
An angle θ between the first direction and the first direction is 0 <θ in a microchip configured by connecting the second mixing tank, the microchannel, and the detection unit in series in this order. A method of using the microchip by sequentially applying a centrifugal force in the second direction of <180 °,
(B) the centrifugal force in the second direction Accordingly, metered sample and / or reagent constitutes the mixture was moved to the mixing tank, depending on the centrifugal force in the first direction, the liquid mixture a second Moving to the mixing tank,
(C) a sample prior to detecting by the detecting unit, depending on the centrifugal force in the second direction, wetting the detector liquid mixture moves to the detector through the microchannel from the second mixing tank Process,
(C1) the step of moving the mixed liquid to the second mixing tank by the centrifugal force in the first direction;
(D) a step of detecting a sample in the detection unit by introducing a liquid mixture by centrifugal force in the second direction into the detection unit that has been wetted by the step of wetting in (c);
Including
Due to the centrifugal force in the second direction, the liquid mixture moves from the second mixing tank through the micro flow path to the detection unit, and the liquid mixture moves from the detection unit to the micro by the centrifugal force in the first direction. A method of using a microchip, characterized in that the second mixing tank, the micro flow path, and the detection unit are arranged at a position where the second mixing tank moves to the second mixing tank . The method of claim 13 , wherein (a) weighing is performed by centrifugal force in the first direction . The method according to claim 13 , wherein (b) mixing and (bb) wetting of the detection unit with a reagent are performed simultaneously. The detection unit is arranged in the second direction with respect to the second mixing tank, and the microchannel connected to the detection unit connects the second mixing tank and the detection unit, and the second mixing tank of the microchannel 16. The device according to claim 13 , wherein the direction from the first to the detection unit is arranged to be greater than −90 ° and less than + 90 ° with respect to the second direction over the entire length of the microchannel. The method described in one. A microchannel formed on a microchip and having the same cross-sectional area over the entire length by connecting the mixing tank and the detection unit ;
A rotation means for sequentially rotating the microchip by applying a centrifugal force in a second direction in which an angle θ between the first direction and the first direction is 0 <θ <180 °. Because
The mixing tank, the microchannel, and the detection unit are configured to be connected in series in this order, and the direction from the mixing channel of the microchannel to the detection unit is the first in the entire length of the microchannel . Is formed in a shape larger than −90 ° and smaller than + 90 ° with respect to the direction of
By rotating the microchip by the rotating means,
(B) introducing the sample and the reagent into the mixing tank by centrifugal force in the second direction, mixing the sample with the reagent in the mixing tank,
(C) Before detecting the sample by the detection unit, the detection liquid is introduced into the detection unit by introducing a mixed solution of the reagent and the sample to be detected by the detection unit by the centrifugal force in the first direction. part to get wet et al.,
(C1) returning the mixed liquid to the mixing tank by centrifugal force in the second direction;
(D) Introducing the mixed liquid by the centrifugal force in the first direction to the detection unit already wetted by (c), and detecting the sample by the detection unit,
Due to the centrifugal force in the first direction, the mixed solution moves from the mixing tank to the detection unit through the micro flow channel, and the mixed solution moves from the detection unit to the micro flow channel by the centrifugal force in the second direction. The mixing tank, the microchannel, and the detection unit are arranged at a position where the mixing tank, the microchannel, and the detection unit are moved through the system . A microchip comprising at least a microchannel having the same cross-sectional area over the entire length by connecting the mixing tank formed on the substrate, the detection section, and the mixing tank and the detection section ;
A rotation means for sequentially rotating the microchip by applying a centrifugal force in a second direction in which an angle θ between the first direction and the first direction is 0 <θ <180 °. Because
The mixing tank, the microchannel and the detection unit are connected in series in this order,
The direction from the mixing tank of the microchannel to the detection unit is formed in a shape that is greater than -90 ° and less than + 90 ° with respect to the first direction in the entire length of the microchannel,
By rotating the microchip by the rotating means,
(B) introducing the sample and the reagent into the mixing tank by centrifugal force in the second direction, mixing the sample with the reagent in the mixing tank,
(C) Before detecting the sample by the detection unit, the detection liquid is introduced into the detection unit by introducing a mixed solution of the reagent and the sample to be detected by the detection unit by the centrifugal force in the first direction. part to get wet et al.,
(C1) returning the mixed liquid to the mixing tank by centrifugal force in the second direction;
(D) Introducing the mixed liquid by the centrifugal force in the first direction to the detection unit already wetted by (c), and detecting the sample by the detection unit,
Due to the centrifugal force in the first direction, the mixed solution moves from the mixing tank to the detection unit through the micro flow channel, and the mixed solution moves from the detection unit to the micro flow channel by the centrifugal force in the second direction. The mixing tank, the microchannel, and the detection unit are arranged at a position where the mixing tank, the microchannel, and the detection unit are moved through the system . The microchip further has a weighing unit,
When a centrifugal force in the second direction is applied, the sample and / or reagent weighed in the weighing unit moves to the mixing tank
The system of claim 18 you. The system according to claim 19 , further comprising a mixing tank disposed in a second direction with respect to the weighing unit, and a detection unit disposed in the first direction with respect to the mixing tank. 21. The system according to any one of claims 18 to 20 , wherein the microchip is configured by arranging at least one reagent holding part upstream of the mixing tank or in the mixing tank or downstream of the detection part. The microchip further includes a separation unit, and when the centrifugal force is applied in the first direction , the separation unit moves from the separation unit to the measurement unit, but moves to the mixing tank. The system according to any one of claims 18 to 21 , wherein the system is configured to be connected to both the weighing unit and the mixing tank. At least a measurement unit, a mixing tank, a second mixing tank, a detection unit, and a microchannel having the same cross-sectional area over the entire length by connecting the second mixing tank and the detection unit formed on the substrate. Microchip,
Rotating means for rotating the microchip by sequentially applying a centrifugal force in a second direction in which an angle θ between the first direction and the first direction is 0 <θ <180 ° to the microchip. And a system including:
The second mixing tank, the microchannel and the detection unit are connected in series in this order,
By rotating the microchip by the rotating means,
(B) The measured sample and / or reagent moves to the mixing tank by the centrifugal force in the second direction to form a mixed liquid, and the mixed liquid becomes the second mixing tank by the centrifugal force in the first direction. The process of moving to,
(C) The step of wetting the detection unit by moving the mixed solution from the second mixing tank through the microchannel to the detection unit by centrifugal force in the second direction before the sample is detected by the detection unit. ,
(C1) the step of moving the mixed liquid to the second mixing tank by the centrifugal force in the first direction;
(D) introducing the mixed liquid into the already wetted detection unit by the step of wetting in (c) by the centrifugal force in the second direction, and detecting the sample by the detection unit;
Including
Due to the centrifugal force in the second direction, the liquid mixture moves from the second mixing tank through the micro flow path to the detection unit, and the liquid mixture moves from the detection unit to the micro by the centrifugal force in the first direction. system at a position moved to the second mixing tank through the channel, the second mixing vessel, characterized in that the microchannel and the detection unit is disposed. Furthermore, the mixing tank is disposed in the second direction with respect to the measuring unit, the second mixing tank is disposed in the first direction with respect to the mixing tank, and the detection unit is second with respect to the second mixing tank . 24. The system of claim 23 , wherein the system is arranged in the direction of