JP4199609B2 - ANALYSIS CHIP, ANALYSIS CHIP UNIT, ANALYSIS DEVICE, AND METHOD FOR PRODUCING ANALYSIS CHIP - Google Patents

Description

【０２３３】
表１より、凸状部材を有する分析用チップでは、従来の分析用チップに比べてドリフト量が小さいことが分かる。これより、凸状部材を有する分析用チップでは、精度の高い分析を行なうことができ、また、分析作業のやり直し頻度及びそれに伴う液体検体の使用量を小さくすることができるので、分析を効率的に行なえることが確認された。
【０２３４】
【発明の効果】
以上詳述したように、本発明の分析用チップ、分析用チップユニット、及び、分析装置によれば、液体検体を効率的に且つ精度良く分析することができるほか、次のいずれかの効果を得ることができる。
（１）液体検体の周り込みによる気泡の発生を抑制することができる。
（２）分析用チップの変形を防止することができる。
（３）液体検体の少量化を行なうことができる。
【０２３５】
また、本発明の分析用チップの作製方法によれば、中間プレートの基板とは反対側の面が、中間プレートの内部孔の壁面及び／又は基板の流路側表面よりも、特定物質含有液に対する親和性が低い部材により構成され、複数のスリット状孔が形成された中間プレートを基板上に固定し、次いで、中間プレートのスリット状孔を通して基板に特定物質含有液を滴下し、特定物質を基板にスポット状に固定させた後、中間プレート上に蓋部を固定するので、特定物質含有液が中間プレートの仕切壁にかかってしまったとしても、特定物質含有液は、中間プレートの基板とは反対側の面よりも特定物質含有液に対する親和性が高い内部孔の壁面及び／又は基板の流路側表面を通じて、基板へ流れる（誘導される）。つまり、特定物質を、基板上の目標とする位置に安定して固定することができ、特定物質が固定された分析用チップを簡単に精度良く作製することができる。
【０２３６】
【図面の簡単な説明】
【図１】本発明の第１実施形態としての分析用チップについて示す図であり、（ａ）はその模式的な組立斜視図、（ｂ）はその模式的な分解斜視図である。
【図２】本発明の第１実施形態としての分析用チップについて示す模式的な図であり、（ａ）は図１（ａ）のＹ−Ｙ断面図、（ｂ）は図１（ａ）のＸ１−Ｘ１断面図、（ｃ）は図１（ａ）のＸ２−Ｘ２断面図である。
【図３】本発明の第１実施形態としての分析用チップについて示す模式的な図であり、（ａ）はその蓋部の上面図、（ｂ）はその中間プレートの上面図、（ｃ）はその中間プレートの上面図、（ｄ）はその基板の上面図である。
【図４】液体検体の流れ方向の定義を説明するための模式図である。
【図５】本発明の第１実施形態としての分析用チップの作製方法について説明するための模式的な上面図である。
【図６】（ａ）は従来のシート形状の空間内に形成された流路を模式的に示す平面図、（ｂ）は本発明の第１実施形態としての分析用チップのスリット状流路を模式的に示す平面図である。
【図７】本発明の第１実施形態の第１変形例を示す模式的な図であり、（ａ）は蓋部の下面図、（ｂ）は中間プレートの上面図、（ｃ）は基板の上面図である。
【図８】本発明の第１実施形態の第２変形例を示す図であり、（ａ）は蓋部の下面図、（ｂ）は基板の上面図である。
【図９】本発明の第１実施形態の第３変形例を示す模式的な図であり、（ａ）は蓋部の上面図、（ｂ）は中間プレートの上面図、（ｃ）は基板の上面図である。
【図１０】本発明の第２実施形態にかかるＳＰＲセンサの全体構成を示す模式的な斜視図である。
【図１１】本発明の第２実施形態としての分析用チップの構成を示す模式的な分解斜視図である。
【図１２】本発明の第３実施形態としての分析用チップについて示す図であり、（ａ）はその模式的な組立斜視図、（ｂ）はその模式的な分解斜視図である。
【図１３】本発明の第３実施形態としての分析用チップにそなえられる中間プレートを示す模式的な下面図である。
【図１４】本発明の第３実施形態としての分析用チップについて示す模式的な図であり、（ａ）は図１２（ａ）のＹ１−Ｙ１断面図、（ｂ）は図１２（ａ）のＸ３−Ｘ３断面図である。
【図１５】本発明の第３実施形態としての分析用チップの第１変形例について示す模式的な図であり、（ａ）は蓋部の下面図、（ｂ）は基板の上面図である。
【図１６】本発明の第３実施形態としての分析用チップの第２変形例について示す模式的な図であり、（ａ）は蓋部の下面図、（ｂ）は基板の上面図である。
【図１７】（ａ），（ｂ）は、本発明の第３実施形態としての分析用チップの第３変形例を示す模式的な断面図である。
【図１８】本発明の第３実施形態にかかる中間プレートの変形例を示す模式的な下面図である。
【図１９】本発明の第４実施形態にかかる分析用チップを示す模式的な図で、（ａ）は中間プレートの下面図、（ｂ）は蓋部の下面図、（ｃ）は基板の上面図である。
【図２０】本発明の第５実施形態にかかる中間プレートを示す模式的な図で、（ａ）は中間プレートの下面図、（ｂ）は（ａ）のＸＸｂ部を拡大して示す図、（ｃ）は従来のプレートの要部を拡大して示す図である。
【図２１】本発明の第６実施形態にかかる分析用チップを示す模式的な図であり、（ａ）は分析用チップの流路の、仕切壁が形成された部分を流路の幅方向に直交する面で切断した断面図、（ｂ）は（ａ）のＸＸＩｂ部を拡大して示す図である。
【図２２】本発明の第７実施形態にかかる中間プレートを示す模式的な下面図である。
【図２３】（ａ），（ｂ）はそれぞれ本発明の第７実施形態を説明するための模式図である。
【図２４】本発明の第７実施形態の第１変形例にかかる中間プレートを示す模式的な下面図である。
【図２５】本発明の第７実施形態の第２変形例にかかる中間プレートを示す模式的な下面図である。
【図２６】（ａ），（ｂ），（ｃ）はそれぞれ、本発明の第７実施形態の第３〜第５変形例にかかる中間プレートを示す模式的な下面図である。
【図２７】本発明の第８実施形態の分析用チップユニットの模式的な斜視図である。
【図２８】本発明の第８実施形態の変形例の分析用チップユニットの模式的な斜視図である。
【図２９】本発明の第９実施形態の分析用チップユニットを説明する模式的な平面図である。
【図３０】本発明の第１０実施形態にかかる分析装置を説明する模式的な図である。
【図３１】本発明の第１０実施形態の変形例にかかる分析装置を説明する模式的な図である。
【図３２】本発明の実施形態を示す模式的な図であり、（ａ）は蓋部を示す上面図、（ｂ）は第１のプレートを示す上面図、（ｃ）は第２のプレートを示す上面図、（ｄ）は基板を示す上面図である。
【図３３】本発明の実施形態にかかる分析用チップを示す模式的な図で、（ａ）はその分解斜視図、（ｂ）は要部を流路の流れ方向に直交する面で切断した断面図である。
【図３４】本発明の実施形態を示す、分析用チップの模式的な組み立て斜視図である。
【図３５】（ａ）〜（ｆ）はいずれも本発明の分析用チップの流路の例を説明するための模式的な図である。
【図３６】（ａ），（ｂ）はいずれも従来の分析用チップを説明するための模式的な図である。
【図３７】従来の分析用チップを説明するための模式的な図である。
【図３８】（ａ），（ｂ）は従来の分析用チップを説明するための模式的な図である。
【図３９】（ａ），（ｂ）は従来の分析用チップを説明するための模式的な図である。
【図４０】液体検体が先回りする様子を説明する図である。
【図４１】液体検体の先回りを防止するための流路の構成を説明する図である。
【図４２】液体検体の先回りを防止するための流路の構成を説明する図である。
【図４３】液体検体の先回りを防止するための流路の構成を説明する、流路の横断面図である。
【図４４】液体検体の先回りを防止するための流路の構成を説明する図である。
【図４５】本発明の第１実施形態の変形例を示す図で、（ａ）はその蓋部の上面図、（ｂ）はそのプレートの上面図、（ｃ）はその基板の上面図である。
【図４６】本発明の実施例を説明するための図である。
【図４７】本発明の実施例を説明するための図である。
【図４８】（ａ）は本発明の実施例の結果を示すグラフ、（ｂ）は従来の分析用チップを用いた結果を示すグラフである。
【符号の説明】
１，１Ａ，１Ｂ，１Ｃ，１Ｃ，１Ｅ，１Ｆ 分析用チップ
１Ｇ，１Ｈ 分析用チップユニット
２ 蓋部
２ａ スリット状溝
２ｂ 仕切壁
４ 基板
４ａ スリット状流路
４ｂ 仕切壁
５，５′ 流路
５ａ 流路５の横断面の長辺
５ｂ 流路５の横断面の短辺
５ｘ 疎水部（第２の親和部）
５ｙ 親水部（親水性部。第１の親和部）
６ 反応部
８，８′ プレート（中間プレート）
９，９′ プレート（中間プレート）
８ａ′，９ａ，９ａ′ スリット状孔（スリット状流路）
８ｂ′，９ｂ，９ｂ′ 仕切壁
１０ プレート（中間プレート）
１０ａ スリット状溝（スリット状流路）
１０ｂ 仕切壁
１０ｃ 縮流部
１０ｔ ４フッ化エチレンの層
１１，１２ 孔
１３，１４ 凹部
２１ 孔（注入口）
２１′，２２′ 溝部（集合流路部）
２６，２７ パイプ
２２ 孔（排出口）
２３，２４ 溝部
４１ 金属層
４２ 回折格子
４３，４４，４６，４７ 溝部（凹部）
５１ 凸状部材
５２，５２ａ〜ｅ 内部流路
６１ 特定物質
８１ 孔（集合流路部）
８１ｘ 孔８１の上流端部
８２ 孔（集合流路部）
８２ｘ 孔８２の下流端部
８３ スリット状孔
９１ スリット状孔（スリット状流路）の上流端部
９２ スリット状孔（スリット状流路）の下流端部
９１′，９２′ 孔
１００ 光源
１０１ 検出器
２００ 気体
２０１ 気泡
２０２ 粒子状物質
２０４ 反応領域
２０４ａ 気泡２０１の近辺
２０５ 測定領域
３００，４００ ユニットベース
３０２ 回転支持軸
３０１，４０１ 単位チップ
４０２ 連結流路
５０１ 分析部
５０２ 分離装置
５０３ 後分析装置
６０１ シリコンウェハー
６０２ Ｕ字形状の型
６０３ 樹脂平板
Ｆｓ 液体検体
Ｓ、Ｓｔ１〜Ｓｔ４ 固−気−液三相境界線
Ｗ 流路の幅
Ｈ 流路の高さ
Ｐ１ スリット状構造を有するパターン
Ｐ２ スリット状構造を持たないパターン[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a chip for analyzing a liquid sample by flowing the liquid sample through a flow channel, a chip unit, an analysis apparatus using the chip, and a method for fixing the chip manufacturing method.
[0002]
[Prior art]
Conventionally, various techniques for detecting a reaction or binding between a liquid specimen and a substance immobilized on a chip and quickly performing a plurality of analyzes in a fixed time have been proposed and put into practical use as a high-throughput analysis system. Yes.
As such a technique, in recent years, a circulation type analysis chip (microchannel chip) such as a DNA chip or a protein chip has attracted attention.
[0003]
In the microchannel chip, a channel having a minute cross section is formed in the chip body, and a substance (specific substance) that interacts with the predetermined chemical substance described above on the wall surface forming the channel. ) Is fixed. A liquid sample is circulated through this flow path, passed through a specific substance on the wall surface of the flow path, or temporarily stopped on the specific substance, thereby bringing the liquid specimen and the specific substance into contact with each other, so that a predetermined amount in the liquid sample is obtained. If a chemical substance (measuring object) is included, this can be detected as an interaction of a specific substance.
[0004]
In addition, as a technique for immobilizing a specific substance on a chip at a high density in a DNA chip or a protein chip, for example, a spotter [Affymetrix 417 (registration) that holds an immobilization target object (specific substance) on a pin tip. (Trademark) Arrayer, etc.) and those in which an immobilization target is sprayed onto a chip by inkjet or a dispenser (Tango (registered trademark) Liquid Handling System, etc.).
[0005]
In addition, if an analysis method based on SPR (Surface Plasmon Resonance) (for example, Biacore (registered trademark)) is combined with such a microchannel chip, the process of binding and dissociating between the measurement object and the specific substance is online. Can be detected.
[0006]
Now, many liquid samples analyzed using a microchannel chip have a limited use amount. For example, in a DNA chip or a protein chip, a liquid sample is a substance collected from various organisms or various substances (DNA, RNA, PNA, peptide, protein, etc.) synthesized biochemically and can be collected. Since the amount is often limited or a large amount of labor is required for collection, there is a strong desire to keep the amount used as small as possible.
[0007]
Analytical chips such as DNA chips and protein chips as described above usually have a large number of specific substances arranged in a flat shape on the bottom surface of the flow path, and then a liquid sample is flowed through the flow path and comes into contact therewith. It has a configuration to let you. In order to perform an analysis efficiently using such an analysis chip, many specific substances are immobilized on one analysis chip. Therefore, the reaction region to which the specific substance is immobilized is localized. It does not become a thing and occupies a relatively large area.
[0008]
Therefore, normally, in order to fix many specific substances, the bottom surface of the flow path is formed to have a large area. Further, at this time, the liquid specimen that does not come into contact with the specific substance diffuses due to the concentration distribution over time, and as a result, may come into contact with the specific substance, but in that case, it may take a long time to contact. Therefore, in order to make the liquid specimen flowing through the flow path efficiently contact with the specific substance fixed to the bottom face of the flow path, that is, the volume of the liquid specimen that does not come into contact with the specific substance fixed to the bottom face of the flow path is reduced. In order to reduce the number, the height (or depth) of the flow path is generally reduced. Therefore, the dimension ratio (long side dimension / short side dimension), which is the ratio between the width and height of the flow path, becomes very large, and the shape of the flow path of the analysis chip is large in the width direction and small in the height direction. It becomes a shape. By making the flow path into a sheet like this, even if the amount of liquid fluid is small, it can be brought into contact with a large number of specific substances through a single flow to improve the analysis throughput, and the analysis can be performed efficiently. It is.
[0009]
In recent years, a number of techniques for such distribution type analysis chips have been proposed. For example, Non-Patent Document 1 discloses a chip that includes a substrate and a sheet member provided with a plurality of slits in parallel. In this technique, when the sheet member is set on the substrate, the slits arranged in parallel function as parallel flow paths on the substrate.
[0010]
Then, different fluids are circulated through the parallel flow paths, the fluids are fixed to the bottom face of the flow path, that is, the substrate, the orientation of the sheet member is changed, and the sheet member is set again on the substrate. A parallel flow path is formed on the substrate so as to intersect the previous parallel flow path. Different fluids are circulated through the flow paths and brought into contact with the fluid previously fixed to the substrate. That is, it is intended to realize a high density (integration) of analysis by forming a joint portion formed by a combination of a large number of fluids on a single chip in a matrix.
[0011]
Further, Patent Document 1 discloses a chip for allowing different liquid specimens to flow through flow paths arranged in parallel and analyzing a plurality of types of liquid specimens simultaneously using one microreactor chip.
[0012]
[Patent Document 1]
WO00 / 04390
[Non-Patent Document 1]
Anal. Chem. 73, 22, pp. 5525, 2001
[0013]
[Problems to be solved by the invention]
In the conventional microchannel chip described above, since the liquid specimen is generally circulated from the state where the initial gas (mainly air) is filled in the flow path, the solid-gas-liquid is flown in the flow path. The three-phase boundary will move. At that time, since the cross section of the flow path is long in the width direction of the flow path, due to non-uniformity of wettability of the flow path wall surface, vibration of the apparatus, adhesion of dust to the flow path surface, etc., FIG. As shown, the solid-gas-liquid three-phase boundary line (tip of the liquid specimen Fs) S has a non-uniform shape that is not linear in the direction of the flow path width direction B, which is a direction perpendicular to the flow direction A. It will become.
[0014]
Then, from the state shown in FIG. 36A, the leading portion of the solid-gas-liquid three-phase boundary line S goes around as shown by the arrow F along with the flow of the liquid specimen Fs, as shown in FIG. Then, a part of the gas 200 is completely surrounded, and bubbles 201 are formed in the liquid specimen Fs. In addition, in the sheet-shaped flow path, the contact interface area between the bubbles 201 and the flow path wall surface becomes large. Therefore, even if liquid feeding is continued, the bubbles 201 are pushed away in the downstream direction and eliminated. In many cases, the bubbles 201 stay as they are.
[0015]
Although it is conceivable that the bubbles 201 are forced out of the flow path by applying a high pressure to the flow path, depending on the structure, material, and joining method of the analysis chip, the flow path may be damaged due to a sudden rise in pressure. There is a risk of doing. At this time, if the substance constituting the flow path is hard, even if a high pressure is applied to the flow path, the influence of the pressure is small. Further, when the flow path structure is constituted by a combination of a plurality of materials, if the strength of bonding between the materials is high, the influence of the pressure is reduced even when a high pressure is applied to the flow path. Furthermore, when pipes (tubes), connectors, etc. are connected to the flow path, if the strength of the coupling between the pipes (tubes), connectors, etc. and the flow paths is high, even if high pressure is applied to the flow paths The effect of the pressure is reduced. However, if the material, joining method, piping connection, etc. employed in the analysis chip are limited for some reason, the bubbles 201 are forcibly excluded from the flow path by applying high pressure to the flow path. It is assumed that this is not preferable. For example, when an analysis chip is composed of a combination of a plurality of material groups (for example, a combination of metal and glass, or resin and glass), the same material group is used for analysis from the viewpoint of disposal. It is preferable to constitute a chip. In addition, in order to reduce the weight of the analysis chip, it is preferable that the analysis chip is made of a light material such as a resin, or in the case of performing a highly accurate processing method such as injection molding or compression molding from the viewpoint of moldability. For this, it is preferable that the analysis chip is made of resin. In addition, when the analysis chip is manufactured, before the analysis using the analysis chip, or when the analysis chip is used, the analysis chip is made of glass, metal, etc. It is preferable to constitute. As described above, the material constituting the analysis chip is arbitrarily selected according to the requirements of the designer, manufacturer, user, and the like.
[0016]
If bubbles 201 stay in this way, problems such as those shown in FIGS. 37 to 39 occur.
For example, as shown in FIG. 39A, when the bubbles 201 stay in the reaction region 204, the contact between the specific substance immobilized on the reaction region 204 and the liquid specimen is hindered.
Further, as shown in FIG. 39B, if the bubble 201 stays in the measurement region 205, the analysis cannot be performed accurately. In particular, in the case of optical measurement, if the bubble 201 stays in the measurement region 205 in this way, the measurement becomes impossible. Therefore, after the liquid specimen is removed from the flow path, preparation is performed again. Measurement must be resumed, and the efficiency of analysis work may be extremely reduced.
[0017]
In addition, as shown in FIG. 37, the particulate matter 202 in the liquid specimen Fs specifically aggregates and accumulates in the vicinity of the upstream side of the bubble 201, affecting the subsequent distribution process, mixing process, and reaction process. End up.
[0018]
Further, if the flow around the bubble 201 becomes uneven due to the retention of the bubble 201, in the extreme case, as shown by an arrow G1 in FIG. 38A, a back flow occurs between the upstream side of the bubble 201 and the flow wall surface. And sometimes affect the analysis. Further, under a constant flow rate and a constant pressure, as shown by an arrow G2 in FIG. 38B, the liquid specimen Fs flows along the peripheral surface of the bubble 201 near the bubble 201 and flows faster than the others.
[0019]
For this reason, in the reaction region 204 to which the specific substance is fixed, the number of contacts with the liquid sample molecules is larger in the vicinity 204a of the bubble 201 than in the other regions (the total amount of the liquid sample that passes through is larger). That is, in the reaction region 204, the interaction proceeds under different conditions depending on the position, and accurate analysis of the liquid specimen cannot be performed based on the interaction such as binding or dissociation in the reaction region 204.
[0020]
In addition, if the bubbles 201 stay in the liquid sample, the difference in heat transfer coefficient between the liquid sample Fs and the bubbles 201 may cause temperature non-uniformity in the measurement system, which may affect the analysis result.
[0021]
Techniques related to the suppression of bubbles in the liquid specimen Fs are disclosed in, for example, Patent Documents 2 and 3 below.
Patent Document 2 discloses a recording liquid supply pipe for supplying a recording liquid (ink) to an ink jet head. In this technique, by providing lyophilicity to the inner surface of the supply pipe, air bubbles are reduced. Adhesion to the inner surface of the supply pipe is suppressed, and even if bubbles are attached to the inner surface of the supply pipe, the bubbles are immediately detached from the inner surface of the supply pipe and can be removed by circulation of the liquid in the supply pipe.
However, this technique does not target a sheet-shaped flow path having a small cross section, and cannot prevent the liquid sample from entering the flow path.
[0022]
Patent Document 3 discloses a flow-through sampling cell in which adhesion of bubbles to the corner portion can be suppressed by rounding the corner portion of the flow channel through which the liquid specimen flows.
However, this technique does not suppress the generation of bubbles due to the surrounding of the liquid specimen, and the suppression of the adhesion of bubbles is limited to the corners, and is not very effective.
[0023]
[Patent Document 2]
JP 2001-162817 A
[Patent Document 3]
Japanese National Patent Publication No. 11-508360
[0024]
In addition, the technique described in Non-Patent Document 1 does not form a sheet-like flow of a liquid specimen, and binds the liquid specimen to a large number of specific substances by a single flow, but a solid-gas-liquid three-phase boundary. No attention is paid to the generation of bubbles due to wrapping around the line.
[0025]
Furthermore, the technique described in Patent Document 1 is similar to the technique described in Non-Patent Document 1 described above, in which a flow of a sheet shape is formed by a liquid specimen and a large number of specific substances are brought into contact with one type of liquid specimen. It does not focus on the generation of bubbles due to the surrounding of the liquid specimen, not the technology.
[0026]
In addition, as described above, in analysis chips such as DNA chips and protein chips, a large number of specific substances are arranged in a planar shape and then brought into contact with a liquid sample. Rather than a relatively large area. In general, an analysis chip has a sheet-like shape in which the flow path is large in the width direction and small in height in response to a demand for a small amount of liquid specimen. In such a case, pressing or pulling from the thickness direction of the chip, which works for holding the analysis chip, or compressive stress or tensile stress due to the pressure of the liquid specimen flowing through the flow path in the analysis chip. For this reason, the shape of the analysis chip may be deformed from an ideal initial shape (usually a rectangular parallelepiped shape or a shape close thereto), and a distribution in the width direction related to the height of the flow path may occur. The ideal initial shape is typically a rectangular parallelepiped, but may have a partially curved portion or an inclined straight portion. Absent.
[0027]
This shape deformation is conspicuous when the above dimensional ratio (= long side dimension / short side dimension) is large, and when the strength of the analysis chip material (various elasticity coefficients) is weak.
When the analytical chip is deformed from the ideal initial shape, the height of the flow path relative to the width direction of the flow path is compared to the case where the analytical chip maintains the ideal initial shape. Therefore, the flow velocity distribution of the liquid specimen flowing through the flow channel is changed from the initial flow velocity distribution. When optical analysis is performed, the optical path length of the light passing through the flow path changes from the initial optical path length due to deformation of the analysis chip, or a trace amount of the portion of the analysis chip that transmits the light. The direction of the optical axis may change due to a slight displacement. In addition, when performing analysis by reflecting light on the surface of the analysis chip or the surface of the flow channel in optical analysis, the position of the optical axis changes compared to the initial ideal shape. End up. As described above, if the flow velocity of the liquid specimen, the optical path length of light, the direction and position of the optical axis, and the like change, there is a possibility that accurate analysis cannot be performed.
Furthermore, the analysis chip's retention force or liquid may change as a result of changes in the type, flow rate, pressure, etc. of the sample liquid, the retention force of the analysis chip, or changes in humidity, temperature, etc. When the shape deformation of the analysis chip is accompanied by temporal fluctuations due to changes in characteristics such as the viscosity of the specimen over time, there is a possibility that accurate analysis may be hindered.
[0028]
The present invention has been devised in view of such a problem. An analysis chip, an analysis chip unit, an analysis apparatus, and an analysis chip that enable efficient and accurate analysis of a liquid specimen are disclosed. An object is to provide a manufacturing method.
[0029]
[Means for Solving the Problems]
As a result of earnest research, the inventor of the present invention circulates a liquid specimen in the flow path, and is fixed on the predetermined substance, facing the flow path, and based on the interaction between the predetermined substance and the specific substance. In the analysis chip used to perform the analysis relating to the liquid specimen, a convex member is formed in the flow path, thereby reducing the amount of the liquid specimen used and making the analysis of the liquid specimen efficient. And the present invention was completed.
[0030]
  In other words, the analysis chip of the present invention allows a liquid specimen to flow through a flow path having a closed cross-sectional structure by a pressure-type transport means, and allows a predetermined substance and a specific substance fixed facing the flow path to interact with each other. An analysis chip used for performing an analysis on the liquid specimen based on an action, wherein the flow path has a convex member.The
[0031]
  In addition, the flow path is preferably formed in a sheet-like space.Yes.
  Further, the analysis chip is provided at the upstream end of the flow channel, and is provided with one inlet for injecting the liquid sample and the downstream end of the flow channel for discharging the liquid sample. It is preferable to have two outletsYes.
[0032]
  In the analysis chip, the convex member is configured as a partition member that divides the flow path in the width direction, and the flow path has a plurality of internal flow paths divided by the partition member. Prefer to beYes.
[0033]
  Further, the analysis chip is interposed between the substrate, the lid member, and the substrate and the lid member, and has a sheet shape having the flow path in cooperation with at least one of the substrate and the lid member. It is preferable to be configured to include at least one intermediate plate that forms a space.Yes.
[0034]
  In the analysis chip, one or more internal holes are formed in the intermediate plate, the substrate and the lid member are overlapped with the intermediate plate interposed therebetween, and the internal flow path is formed by the internal holes. Preferably to beYes.
[0035]
  Further, in the analysis chip, the surface of the intermediate plate opposite to the substrate is more compatible with the liquid containing the specific substance than the wall surface of the inner hole of the intermediate plate and / or the surface of the substrate on the flow path side. It is preferable that it is composed of a low materialYes.
[0036]
  Further, the analysis chip is configured to include a substrate and a lid member that is disposed to face the substrate and forms a sheet-like space having the flow path in cooperation with the substrate. LikeYes.
[0037]
  Further, the analysis chip is configured such that the substrate and the lid member overlap each other, and the internal flow path is formed on at least one of the opposing surfaces of the substrate and the lid member. Is preferredYes.
[0038]
  In addition, it is preferable that the analysis chip has a constricted portion at the downstream end of the internal flow path where the internal flow path gradually narrows.Yes.
[0039]
  In the analysis chip, the internal flow path is preferably formed from the inlet to the outlet.Yes.
[0040]
  In the analysis chip, the partition member is configured as a partition wall, and the internal channel is a slit-like channel divided by the partition wall at an intermediate portion in the flow direction of the channel. It is preferable to have a collecting flow path portion that is formed at the upstream end portion and the downstream end portion in the flow direction of the flow path and collects the liquid specimen.Yes.
[0041]
  The analysis chip is formed such that the collecting channel on the upstream end side becomes wider from the inlet to the intermediate portion, and the collecting channel on the downstream end side is formed. The part is preferably formed so as to become narrower as it goes from the intermediate part to the outlet.Yes.
[0042]
  In the analysis chip, it is preferable that each of the collecting flow path portions on the upstream end side and the downstream end side is provided on the substrate or the lid member.Yes.
[0043]
  The analysis chip has a slit-like flow path of 5 mm.²It is preferable to have the following cross-sectional areaYes.
[0044]
  Further, the analysis chip preferably has an aspect ratio of the cross section of the slit-shaped channel of about 0.005 to 100.Yes.
[0045]
  In the analysis chip, it is preferable that the specific substance is fixed at a plurality of spots in the form of spots facing the internal flow path and spaced apart from each other.Yes.
[0046]
  In the analysis chip, the convex member is preferably a support member interposed between the opposing surfaces of the flow path.Yes.
[0047]
  Further, the analysis chip is interposed between the substrate, the lid member, and the substrate and the lid member, and has a sheet shape having the flow path in cooperation with at least one of the substrate and the lid member. At least one intermediate plate that forms a space, and the column member is disposed between the mutually facing surfaces of the intermediate plate and at least one of the substrate and the lid member in the flow path in the sheet-like space. It is preferable to be intervenedYes.
[0048]
  The analysis chip includes a substrate and a lid member that is disposed to face the substrate and forms a sheet-like space having the flow path in cooperation with the substrate, It is preferable that the flow path in the sheet-like space is interposed between the mutually facing surfaces of the substrate and the lid member.Yes.
[0049]
  The analysis chip forms the sheet-like space on the left side surface, the right side surface, the upstream end surface, and the downstream end surface in addition to the floor surface and the ceiling surface constituting the flow path, and supports the support. It is preferable that the member is interposed between at least one of the left side surface and the right side surface and between the upstream end surface and the downstream end surface.Yes.
[0050]
  In the analysis chip, it is preferable that the support member is in direct contact with the facing surface.Yes.
[0051]
  In addition, when the analysis chip has a part of the support member directly in contact with one of the opposing surfaces and the other end of the support member circulates fluid through the flow path, It is preferable to contact the other of the opposing surfaces via the fluid.Yes.
[0052]
  In addition, the analysis chip preferably has an adhesion reducing layer formed on the surface of the support member.Yes.
[0053]
  Further, the analysis chip preferably has the specific substance fixed to the flow path.Yes.
[0054]
  In the analysis chip, the flow path is provided with a first affinity part and a second affinity part having a lower affinity for the liquid specimen than the first affinity part. LikeYes.
[0055]
  In the analysis chip, the specific substance is immobilized on the surface of the flow path, and the first affinity portion and the upstream of the portion where the specific substance is immobilized in the flow direction of the flow path. It is preferable to have a second affinity partYes.
[0056]
  In the analysis chip, it is preferable that the first affinity part and the second affinity part are each formed in a strip shape extending in a direction intersecting the flow direction of the flow path.Yes.
[0057]
  In the analysis chip, it is preferable that the first affinity part and the second affinity part are formed alternately and in a plurality.Yes.
[0058]
  In addition, it is preferable that the first affinity part is a hydrophilic part and the second affinity part is a hydrophobic part.Yes.
  The first affinity part is preferably a rough surface part, and the second affinity part is preferably a smooth surface part.Yes.
[0059]
  The analysis chip includes a surface on which the specific substance is fixed in the flow path, a diffraction grating that generates an evanescent wave by light irradiation on the surface, and a metal layer that can induce a surface plasmon wave. Is preferredYes.
[0060]
  Further, the analysis chip is preferably made of a material having a Young's modulus of 60 GPa or more and 1000 GPa or less.Yes.
[0061]
  Another analysis chip of the present invention includes a specific substance in a flow path, and a liquid specimen is circulated through the flow path by a pressure-type transporting means, so that the predetermined substance and the specific substance in the liquid specimen are mutually exchanged. An analysis chip used for performing an analysis on the liquid specimen based on the action includes one inlet for injecting the liquid specimen and one outlet for discharging the liquid specimen. The road has a cross section with an aspect ratio of about 0.005 to 100, and 5 mm.²It has the following cross-sectional area and is provided in parallel between the inlet and the outlet.The
[0062]
  The analysis chip unit of the present invention has a unit base having a plurality of surfaces, and the analysis chip is provided as a unit chip on the surface of the unit base.The
[0063]
  Another analysis chip unit of the present invention includes a unit base, and includes a plurality of the above-described analysis chips as unit chips on the unit base, and the corresponding unit chips among the plurality of unit chips are connected. A connection channel is provided.The
[0064]
  The method for producing an analysis chip according to the present invention is a method for producing an analysis chip, wherein the specific substance is immobilized on the analysis chip, and an analysis chip having the specific substance immobilized thereon is produced. The intermediate plate is fixed to the substrate, then the specific substance-containing liquid is dropped onto the substrate through the internal hole of the intermediate plate, and the specific substance is fixed to the substrate in a spot shape. The lid is fixed.The
[0065]
  An analysis apparatus according to the present invention includes the analysis chip or the analysis chip unit, and an analysis unit that analyzes a liquid sample.The
[0066]
  At this time, the analysis unit performs analysis by an analysis method using at least one method selected from the group consisting of surface plasmon resonance, chemiluminescence, bioluminescence, electrochemiluminescence, fluorescence, and radioisotope analysis. Preferably to doYes.
[0067]
  Further, the analyzer preferably includes a separation unit that separates the liquid sample by physical and / or chemical action prior to introducing the liquid sample into the analysis chip or the analysis chip unit. BetterYes.
[0068]
  In addition, the analyzer preferably includes a post-analysis unit that analyzes the liquid sample discharged from the analysis chip or the analysis chip unit.Yes.
[0069]
DETAILED DESCRIPTION OF THE INVENTION
Embodiments of the present invention will be described below with reference to schematic drawings. In each of the following embodiments, an example is described in which a liquid sample is water-soluble (hydrophilic, here the solvent is water), but the liquid sample is hydrophobic. Needless to say, a liquid sample can be measured using the analysis chip of the present invention. The liquid specimen referred to in the present invention is a substance that can cause interactions such as antigen-antibody reaction, complementary DNA binding, receptor / ligand interaction, enzyme / substrate interaction, and the like. For example, proteins, nucleic acids, DNA, RNA, PNA, peptides, hormones, antigens, antibodies, ligands, receptors, enzymes, substrates, low molecular weight organic compounds, cells, ions, and complexes thereof It is a liquid containing (or possibly containing), and also includes dispersion systems such as suspensions and colloids. These may be labeled with a fluorescent substance, a luminescent substance, a radioactive substance, or the like as necessary.
[0070]
[1] First embodiment
1 to 3 show an analysis chip as a first embodiment of the present invention. FIG. 1 (a) is a schematic assembly perspective view thereof, and FIG. 1 (b) is a schematic exploded perspective view thereof. 2A is a sectional view taken along the line YY in FIG. 1A, FIG. 2B is a sectional view taken along the line X1-X1 in FIG. 1A, and FIG. 2C is a sectional view taken along the line X2 in FIG. FIG. 3A is a top view of the lid, FIG. 3B is a top view of the first plate, FIG. 3C is a top view of the second plate, FIG. (D) is a top view of the substrate. The flow direction A of the liquid specimen Fs referred to below is the main flow direction in the flow path. For example, in the flow path 5 ′ as shown in FIG. 4, the flow direction is indicated by a solid arrow. It shall refer to the direction.
[0071]
As shown in FIGS. 1 (a) and 1 (b), the analysis chip (also simply referred to as a chip) 1 includes a lid portion 2 that is a flat lid member and a thin first plate (hereinafter simply referred to as a “thin plate”). 8, a second plate 9 having a small thickness (intermediate plate, hereinafter simply referred to as a plate) 9, and the substrate 4. Then, at the time of analysis, these members 2, 8, 9, and 4 are stacked from above in this order and assembled together by a joining holder (not shown) as shown in FIG. Therefore, the plates 8 and 9 are interposed between the lid portion 2 and the substrate 4.
[0072]
The holder is preferably provided with a protective mechanism for alignment and prevention of scratches. As an example of the protection mechanism, for example, a locking portion provided in the holder for locking the analysis chip 1 or a portion (reaction unit 6 described later) of the analysis chip 1 that is observed does not contact the holder. Examples include a recess formed in the holder.
[0073]
As shown in FIG. 2A, the liquid specimen Fs injected from a hole 21 (an inlet at the upstream end of the flow channel 5) 21 described later is a plate 8 serving as an upstream collecting flow channel. Through the holes 81, flow through the slit-like holes 9 a serving as the internal flow paths of the plate 9, and then pass through the holes 82 of the plate 8, which is the downstream collecting flow path section, to pass through the holes of the lid 2 described later. (Exhaust port at the downstream end of the flow path 5) 22 flows out and is fixed to the reaction part 6 of the substrate 4 when the liquid specimen Fs flows through the slit-like hole 9 a of the plate 9. Each specified substance 61 is brought into contact.
[0074]
Further, as shown in FIGS. 2B and 2C, the flow path 5 through which the liquid specimen Fs circulates is a slit-shaped section that is elongated in the horizontal direction (a section that is perpendicular to the flow direction A of the liquid specimen Fs). ), That is, a channel having a closed cross-sectional structure formed in a sheet-like space.
Here, the “flow path formed in the sheet-like space” as used in the present invention is usually a long side of the cross section (of the cross section perpendicular to the flow direction of the flow path 5 and the cross section perpendicular to the width direction). The longest side is generally referred to as the width or the length in the flow direction of the flow path 5. In this embodiment, the dimension W of 5a is in the range of 500 μm to 100 mm, and The dimension H of the short side (height of the flow path 5) 5b of the cross section is in the range of 5 μm to 2 mm. Further, the range of the dimensional ratio (= long side dimension W / short side dimension H) between the long side 5a and the short side 5b is usually 1.5 or more, preferably 10 or more, and usually 20000 or less, preferably 100 or less. At this time, when the flow path 5 is divided into a plurality of internal flow paths 9a by a partition wall 9b that functions as a convex member, a partition member, and a support member as will be described later, the divided original flow path 5, that is, the dimension of the flow path 5 including all of the plurality of internal flow paths 9a only needs to be within the range of the above dimensional ratio. Here, the length W of the long side 5a is 20 mm, the length of the short side 5b (the thickness H of the plate 8 in the X1-X1 cross section).₁, The thickness H of the plate 9 in the X2-X2 cross section₂) Are both set to 250 μm.
[0075]
In addition, the closed cross-sectional structure here refers to a structure in which a cross section perpendicular to the flow direction of the flow path 5 is a closed cross section. In addition, even when the bottom surface, ceiling surface, or wall surface of the channel 5 is formed of a material having minute pores such as a membrane filter or a gas permeable membrane, When the liquid specimen Fs flowing through the flow path 5 does not pass through the minute pores, the flow path 5 is assumed to have a closed cross-sectional structure. In the present specification, the flow path 5 will be described as a flow path having a closed cross-sectional structure formed in a sheet-like space unless otherwise specified.
[0076]
Hereafter, each said member which comprises this analysis chip | tip 1 is demonstrated in detail.
The material of the lid part 2, the plate 8, the plate 9, and the substrate 4 is not particularly limited, such as resin, ceramics, glass, metal, etc., but the binding between the detection species and the specific substance 61 is caused by fluorescence, light emission, and color development. When the optical measurement is performed using phosphorescence or the like, the lid 2 and the plates 8 and 9 are preferably formed of a transparent material. However, when the analysis chip 1 can be disassembled and measured, the lid 2 and the plates 8 and 9 do not require transparency. Examples of the transparent material include resins such as acrylic resin, polycarbonate, polystyrene, polydimethylsiloxane, and polyolefin, and glass such as Pyrex (registered trademark: borosilicate glass) and quartz glass.
[0077]
In addition, the analysis chip 1 can prevent deformation even with a material having a low strength (Young's modulus is 1 GPa or more and 60 GPa or less) by the configuration of the present embodiment, as described later, but it is configured with a material having a high strength. Therefore, analysis with higher accuracy becomes possible. Therefore, it is preferable that the analysis chip 1, that is, the lid 2, the plate 8, the plate 9, and the substrate 4 are formed of a material having high strength. Specifically, it is preferable that the Young's modulus of the material forming the lid portion 2, the plate 8, the plate 9, and the substrate 4 is usually 60 GPa or more. Here, the analysis chip 1 is formed of a strong material having a Young's modulus of 60 GPa or more.
[0078]
As shown in FIG. 3A, one hole (injection port) 21 is formed at the upstream end of the lid 2, and one hole (discharge port) 22 is formed at the downstream end of the lid 2. Is formed. The inlet 21 is connected to a liquid feed pump (for example, a syringe pump) via a connector and a tube (not shown), and the discharge port 22 is connected to a waste liquid tank via a connector and a tube (not shown). . Then, by operating the liquid feeding pump, the liquid specimen Fs can be injected into the chip 1 from the injection port 21 and discharged from the chip 1.
[0079]
As shown in FIG. 3B, a hole 81 is formed on the upstream side of the plate 8, and a hole 82 is formed on the downstream side of the plate 8.
Further, the upstream end portion 81x of the hole 81 is positioned so as to align and communicate with the injection port 21 of the lid portion 2 when the chip 1 is assembled. Further, the hole 81 is formed to become wider from the upstream end portion 81x toward the intermediate portion in the flow direction of the plate 8 (as it goes downstream in the flow direction of the liquid specimen Fs).
[0080]
On the other hand, the downstream end portion 82x of the hole 82 is positioned so as to align and communicate with the discharge port 22 of the lid portion 2 when the chip 1 is assembled. Further, the hole 82 is formed to become narrower as it goes from the intermediate portion in the flow direction of the plate 8 to the downstream end portion 82x (as it goes downstream in the flow direction of the liquid specimen Fs).
[0081]
When the chip 1 is assembled, the upper and lower surfaces of the plate 8 are closed by the lid portion 2 and the plate 9, and the holes 81 and 82 form a flow path in which the liquid specimen Fs collects. Therefore, the flow path formed by the holes 81 and 82 of the plate 8 is also referred to as the collective flow path portions 81 and 82. In FIGS. 1A and 2, the upper and lower surfaces of the plates 8 and 9 are all closed by the lid portion 2 and the plate 9, but at least the portions where the holes 81 and 82 of the plate 8 are formed are closed. It only has to be done.
[0082]
As shown in FIG. 3 (c), a plurality of slit-like holes (internally divided) formed in the width direction by partition walls (partition members) 9b formed as convex members are formed in the middle portion in the flow direction of the plate 9. (Hereinafter referred to as slit-like hole) 9a is formed. When the chip 1 is assembled, each slit-shaped hole 9a is divided at the middle portion of the flow path 5 by the partition wall 9b to form a slit-shaped internal flow path (hereinafter referred to as a slit-shaped flow path as appropriate). . Here, the internal flow path means a flow path divided in the width direction by the partition member. Therefore, the partition wall 9b is in direct contact with the substrate 4 and the plate 8, and the liquid specimen Fs cannot enter between the partition wall 9b and the substrate 4 and between the partition wall 9b and the plate 8, The flow path 5 is divided into a plurality of internal flow paths. In addition, when the chip 1 is assembled, the upper and lower surfaces of the slit-shaped hole are closed by the plate 8 and the substrate 4 to form a slit-shaped flow path. Since the internal flow path is the same, these are denoted by the same reference numeral 9a.
[0083]
In general, the aspect ratio (vertical dimension / horizontal dimension) of the cross-section of the slit-shaped channel 9a is within a range of about 0.005 (for example, 5 μm in length, 1 mm in width) to 100 (for example, 10 mm in length, 100 μm in width). It is preferable that the slit-shaped flow path 9a is formed so as to fit in the space. In general, the slit-shaped channel 9a is 5 mm.²It is preferable to have the following cross-sectional area. In detail, the cross-sectional area of the slit-shaped channel 9a is usually 100 μm.²Or more, preferably 2000 μm²Above, usually 5mm²Below, preferably 0.3mm²It is as follows.
[0084]
Further, when the chip 1 is assembled, the upstream end portion 91 of each slit-shaped hole 9a is positioned so as to communicate with the downstream end of the hole 81 of the plate 8, and the downstream end portion 92 of each slit-shaped hole 9a. Is positioned so as to communicate with the upstream end of the hole 82 of the plate 8.
Thereby, after the liquid specimen Fs injected into the collective flow path portion 81 of the plate 8 flows through the slit-shaped flow paths 9a of the plate 9 through the upstream end portions 91 of the respective slit-shaped flow paths 9a, the respective slits The plate 8 passes through the downstream end 92 of the channel 9a and gathers into the collecting channel 82 of the plate 8.
[0085]
As described above, in the present analysis chip 1, the flow path 5 is further divided into the minute internal flow paths 9 a by providing the partition wall 9 b in the conventional sheet-shaped flow path 5 (that is, the flow path 5 The cross-sectional area of the liquid sample Fs can be reduced.
[0086]
As shown in FIGS. 1A and 1B, a reaction section 6 is provided in the middle of the flow direction of the substrate 4 so as to face the flow path 5.
The reaction unit 6 is shown in a simplified manner in FIGS. 1 (a) and 1 (b), but as shown in FIG. 3 (d), it interacts with a predetermined substance (detection species) either specifically or non-specifically. The specific substance 61 that acts is fixed to the surface of the substrate 4 on the flow path 5 side at a plurality of spots in a spot shape. At this time, in order to ensure that the specific substance 61 is fixed to the substrate 4, it is desirable that an immobilization film (organic film) that can be bonded to the specific substance 61 is formed on the surface of the substrate 4.
The general range of (vertical dimension × horizontal dimension) of the reaction unit 6 is (3 mm × 3 mm) to (20 mm × 20 mm). Generally, this region has a vertical and horizontal 3 at intervals of 100 μm to 1 mm. A total of 9 to 40,000 specific substances 61 are arranged at 200 pieces each.
[0087]
  Here, as each specific substance 61, a specific substance (specific substance different from each other) that interacts specifically or non-specifically with a reaction or binding is used for different substances. Yes.
  In addition, the predetermined substance and the specific substance are, for example, interactions such as antigen-antibody reaction, complementary DNA binding, receptor / ligand interaction, enzyme / substrate interaction, etc.RawSpecific examples are proteins, nucleic acids, DNA, RNA, PNA, peptides, hormones, antigens, antibodies, ligands, receptors, enzymes, substrates, small organic compounds, cells, and these. And the like. These may be labeled with a fluorescent substance, a luminescent substance, a radioactive substance, or the like as necessary.
[0088]
In addition, as will be described in a method for manufacturing the analysis chip 1 described later, in the analysis chip 1, the plate 9 is fixed on the substrate 4, and then the substrate 4 is passed from above the plate 9 through the slit-like holes 9 a of the plate 9. Since the specific substance 61 is fixed to the substrate 4, the reaction part 6 (the part where the plurality of specific substances 61 are fixed) as shown in FIG. 3D is not formed in the initial stage. In order to explain the arrangement of 61 in an easy-to-understand manner, FIG. 3D shows a state where the specific substance 61 is fixed to the substrate 4 for convenience. Therefore, in FIG. 3D, the position and the number of spots of the specific substance 61 in the width direction are shown to match the positions of the slit holes 9a and the number of slit holes 9a in the intermediate plate 9 in the width direction. .
[0089]
The liquid specimen Fs flowing through the slit-shaped channel 9a comes into contact with these specific substances 61 in the flow process, and the liquid specimen Fs can be analyzed according to the reaction status of each specific substance 61 after the distribution. .
That is, if the reaction of any one of the plurality of specific substances 61 can be observed, it can be detected that a predetermined substance corresponding to the reacted specific substance 61 is included in the liquid specimen Fs.
[0090]
The specific substance 61 is fixed to the chip 1 with a reference interval so as not to cause contamination with the adjacent specific substance 61. Here, the reference interval refers to the interval between the centers of the spots on which the specific substance is fixed, and the pitch of the partition walls 9b is set to be substantially the same as this reference interval. Even if the partition wall 9b is provided, the number of spots per unit area of the specific substance 61 is not reduced more than in the past. On the contrary, since the above-mentioned contamination can be prevented by providing the partition wall 9b, the pitch of the specific substance 61 in the width direction (direction perpendicular to the flow direction) is narrower than before, and the number of spots per unit area is increased. It is also possible to do.
[0091]
Note that it is not always necessary to use different specific substances 61 for each specific substance 61, and the same specific substance 61 may be used. In any case, what specific substance 61 is used is appropriately set according to the purpose of the analysis.
[0092]
Next, a method for producing the analysis chip 1 will be described. First, the plate 9 is bonded onto the substrate 4. Then, as shown in FIG. 5, a dispersion liquid in which the specific substance 61 is dispersed or dissolved in the liquid through the slit-like hole 9 a of the plate 9 by an injector or a spotter (not shown) capable of positioning operation. Alternatively, the specific substance 61 is fixed with a reference interval from each other by dropping a solution. Hereinafter, a dispersion or solution in which the specific substance 61 is dispersed or dissolved in a liquid is referred to as a “specific substance-containing liquid”. The liquid for dispersing or dissolving the specific substance 61 is arbitrary, but in this embodiment, the specific substance-containing liquid is described as an aqueous solution in which the specific substance 61 is dissolved in water. FIG. 5 is a top view schematically showing a state in which the specific substance-containing liquid is dropped by bonding the plate 9 to the substrate 4 and the specific substance 61 is fixed.
Thereafter, the plate 8 is assembled on the plate 9, and the lid portion 2 is assembled on the plate 8.
[0093]
In the method for manufacturing the analysis chip 1 described above, it is preferable that the plate 9 is composed of a member having a lower affinity for the specific substance-containing liquid than the substrate 4. In this embodiment, since the aqueous solution of the specific substance 61 is used as the specific substance-containing liquid, it is preferable to use a hydrophobic member or the like as an example of the member having low affinity. As a result, the surface of the plate 9 on the plate 8 side (opposite side of the substrate 4) has a lower affinity for the specific substance-containing liquid than the surface of the substrate 4 on the flow path 5 side. For this reason, when the specific substance 61 is dissolved or dispersed in the specific substance-containing liquid by the injector and dropped, the dropping position is shifted due to a disturbance or an apparatus, and the specific substance-containing liquid is separated from the partition wall 9b of the plate 9. The specific substance-containing liquid also flows (induces) to the substrate 4 having a higher affinity for the specific substance 61 than the plate 9 even when it is dripped onto the base plate. This is because the specific substance 61 can be stably fixed at a target position on the substrate 4.
[0094]
Further, only the partition wall 9b of the plate 9 is made of a member having a lower affinity for the specific substance-containing liquid than the substrate 4, or the surface of the partition wall 9b of the plate 9 with respect to the specific substance-containing liquid than the substrate 4 is formed. A layer having low affinity may be formed. Further, the plate 8 and the lid portion 2 may be joined and integrated in advance.
[0095]
Further, the surface of the plate 9 on the side of the plate 8 (opposite side of the substrate 4) may be constituted by a member having a lower affinity for the specific substance-containing liquid than the wall surface of the slit-like hole 9a of the plate 9. Thus, in the same manner as described above, even when the specific substance-containing liquid is dropped on the surface of the partition wall 9b of the plate 9 on the plate 8 side surface, the specific substance 61 is separated from the substrate 4 of the plate 9. Flows to the slit-shaped hole 9a having higher affinity for the specific substance-containing liquid than the opposite surface. Thereby, the specific substance-containing liquid can be reliably guided to the slit-shaped hole 9a, and the specific substance 61 can be stably fixed at the target position on the substrate 4.
[0096]
The term “affinity” in the present specification refers to the tendency of a certain substance to bind or adsorb to another substance, and should not be regarded as indicating only hydrophilicity and hydrophobicity. .
[0097]
  In this embodiment, the members are joined by physically combining them with a holder so that the chip 1 can be disassembled. However, the members may be joined by other methods. The bonding method of each member is arbitrary, and examples thereof include bonding with an adhesive, resin bonding with a primer, diffusion bonding, anodic bonding, eutectic bonding, thermal fusion, ultrasonic bonding, laser melting, solvent / dissolving solvent, and the like. However, it may be performed by using an adhesive tape, an adhesive tape, or a self-adsorbing agent, or may be crimped or engaged with each member by providing irregularities. Thereby, assembly can be performed easily. In addition, any combination of these joining methodssoYou may use together.
[0098]
Since the analysis chip according to the first embodiment of the present invention is configured as described above, as shown in FIGS. 2 (a) and 3 (a) to 3 (d), the inlet of the lid portion 2 is used. The liquid specimen Fs injected into 21 flows through the collective flow path portion 81 of the plate 8.
Thereafter, the liquid specimen Fs flows from the upstream end portion 91 of each slit-shaped channel 9 a to each slit-shaped channel 9 a and comes into contact with the specific substance 61.
Then, the liquid specimen Fs gathers from the downstream end portion 92 of each slit-like flow passage 9a to the collecting flow passage portion 82 and is discharged out of the chip 1 through the discharge port 22 of the lid portion 2.
[0099]
Thus, in this analysis chip 1, since one inlet 21 and one outlet 22 are provided for each of the plurality of slit-shaped channels 9 a, a plurality of simply as in the conventional technique described above. Compared to a configuration in which flow paths are provided in parallel and fluids are individually injected and discharged in each flow path, a large number of connectors and tubes used for injection and discharge are not required, and the attachment work of the connectors and tubes to the chip 1 is performed. Is easy.
[0100]
Furthermore, when this analysis chip 1 is used, different types of different liquid specimens Fs can be serially flowed, and these different types of liquids can be continuously measured. Thereby, it is not necessary to prepare the analysis chip 1 for each liquid specimen Fs, and the analysis can be easily performed in a short time.
[0101]
Further, since the collecting flow path portion 81 becomes wider from the upstream end portion 81x to the intermediate portion in the flow direction, the liquid specimen Fs can be smoothly guided to the intermediate portion in the flow direction. Further, since the collecting flow path portion 82 becomes narrower from the intermediate portion in the flow direction toward the downstream end portion 82x, the liquid specimen Fs can be smoothly guided to the downstream end portion 82x.
[0102]
Further, since the partition wall 9b is provided in the flow path 5 formed in the sheet-shaped space to form a finer (narrower) internal flow path (slit-shaped flow path) 9a, the liquid sample Fs is surrounded. Generation of bubbles can be suppressed.
That is, as shown in FIG. 6A, in the conventional sheet-shaped flow path, the solid-gas-liquid three-phase boundary line is long, so that some liquid specimens Fs are caused by non-uniform wettability. As a result, gas entrainment (bubbles 201) due to the entrainment of the liquid specimen Fs occurred, but as shown in FIG. By dividing the channel (slit channel 9a), the line segment (channel width) L perpendicular to the main flow in the channel is reduced, so that the probability of wraparound is greatly reduced. Moreover, since the cross-sectional area of the flow path is reduced, back pressure is efficiently applied to each slit-shaped flow path 9a, and bubbles are less likely to stay.
[0103]
Therefore, according to the present analysis chip 1, adverse effects due to the retention of bubbles (inhibition of the flow of the liquid specimen Fs, inhibition of contact between the specific substance 61 and the liquid specimen Fs, difference in heat transfer coefficient between the liquid Fs and the bubbles 201). The temperature of the measurement system due to the non-uniformity, the measurement interference due to the bubbles 201 remaining in the optical path when performing analysis using light, etc.) can be eliminated, and the reliability of the analysis can be improved. is there. Furthermore, there is an advantage that the work of removing bubbles is not required and the analysis work can be performed efficiently.
[0104]
In the flow path 5 formed in a conventional sheet-shaped space, global flow non-uniformity occurs. That is, in the flow range of the normally supplied liquid fluid, the flow velocity of the liquid fluid on the wall surface is zero, the flow velocity in the center is fast in both the vertical and horizontal directions, and the flow velocity is slow as it approaches the wall surface. Occurs.
However, in the present analysis chip 1, by providing independent minute internal flow paths (slit-shaped flow paths 9a), for example, when the specific substances 61 are provided in two rows in the width direction of the slit-shaped flow paths 9a. Since the period during which the liquid specimen Fs contacts the specific substances 61 arranged in the width direction can be made uniform, the accuracy of the analysis result can be improved.
[0105]
In addition, when the lengths of the main flow of the liquid specimen Fs for each internal flow path 9 a are different, the liquid specimen Fs flowing through the internal flow path 9 a near the center of the flow path 5 flows into the internal flow near the end of the flow path 5. Since the solid-gas-liquid three-phase boundary line precedes the liquid specimen Fs flowing through the path 9a, the liquid specimen Fs from the internal flow path 9a near the center of the flow path 5 in the downstream collecting flow path section It is possible that the process will be “advanced” and cause bubbles to stay in the vicinity of the end of the flow path 5. This is shown in FIG. In FIG. 40, reference numeral 51 denotes a partition wall, and reference numeral 52 denotes an internal flow path. Symbols St1, St2, St3, and St4 indicate solid-gas-liquid three-phase boundaries after a certain time from the start of the flow of the liquid specimen Fs into the flow path 5, and the interfaces are in the order of St1, St2, St3, and St4. Assume that you are moving forward.
[0106]
Accordingly, the length of each internal flow path is made constant to such an extent that “advance” does not occur (see FIG. 41), or first and second affinity portions as described in a seventh embodiment described later are provided. It is desirable to adjust the flow velocity of the liquid specimen Fs flowing through each internal flow path or {see FIG. 26 (c)}. Also, as shown in FIG. 42, it is also desirable to adjust the linear velocity of the liquid specimen Fs so that the cross-sectional area of the internal channel 52 becomes narrower from the center in the width direction of the channel 5 toward the end. Further, by changing the height of the internal flow path 52 to adjust the cross-sectional area (see FIG. 43), adjusting the roughness of the surface of the partition wall 51 and the flow path 5 (see FIG. 44), etc. It is also desirable to adjust the flow rate of the liquid specimen Fs flowing through each internal flow path 9a by adjusting the pressure loss of the liquid specimen Fs flowing through the flow path 9a. However, it is desirable to keep the cross-sectional area constant when there is non-uniform measurement (for example, reaction amount) due to the flow rate of each internal flow path 9a not being constant at the time of measurement. The analysis chip should be optimized according to the reaction species, reaction rate, temperature, flow rate, etc. The configuration shown in FIG. 44 is based on the fact that the pressure loss of the liquid specimen Fs flowing through the flow path 5 is generally increased when the surface of the partition wall 51 or the flow path 5 is rough. 41 to 44, the same reference numerals as those used in FIG. 40 denote the same elements. Also, in FIG. 44, the roughness of the wall surfaces of the internal channels 52a to 52e is adjusted, and the roughness is smoothed in the order of 52a, 52b, 52c, 52d, and 52e.
[0107]
Further, when the chip 1 is assembled by the holder, pressure is applied to the chip 1, but the pressure resistance of the chip 1 can be improved by the partition walls 9 b formed in plural across the chip 1 width direction. Shape change, particularly, a shape change in the thickness direction can be prevented. As a result, nonuniformity in the flow velocity distribution due to the deflection of the chip 1 can be prevented, and in optical analysis, variations in optical path length and changes in the optical axis can be suppressed, so that analysis is performed under optimum conditions. There is an advantage that the accuracy of the analysis result can be improved.
Hereinafter, the embodiment will be described with attention paid to the shape change prevention function.
[0108]
The partition wall 9b of the plate 9 divides the flow path 5 in the width direction. The partition wall 9 b functions as a support member interposed between the opposing surfaces of the flow path 5, that is, the flow path 5 side surface of the substrate 4 and the flow path 5 side surface of the plate 8.
The partition wall 9b connects the substrate 4 and the plate 8 so that the partition wall 9b erected on the substrate 4 supports the plate 8 as a support member, thereby applying a force in the thickness direction of the chip 1. Also, the deformation of the chip 1 can be prevented.
[0109]
Further, the state in which the partition wall 9b is interposed in the flow path 5 as a support member will be described in another expression as follows. That is, the flow channel 5 side surface of the substrate 4 constituting the floor surface of the flow channel 5, the flow channel 5 side surface of the plate 8 constituting the ceiling surface of the flow channel 5, the left side surface, the right side surface of the flow channel 5, The flow path 5 is formed in the sheet-like space by the upstream end face and the surface of the plate 9 facing the slit-like flow path 9a constituting the downstream end face, and the partition wall 9b as a support member is Between the left side surface and the right side surface of the flow path 5 or at least one of the upstream end face and the downstream end face of the flow path 5 (here, between the upstream end face and the downstream end face). It has an intervening configuration.
Therefore, since the upstream end of the flow path 5 and the downstream end of the flow path 5 of the partition wall 9b are connected to the plate 9, the direction in which the partition wall 9b is interposed in the chip 1 {here, the longitudinal direction (flow direction A )} Force can be prevented from being deformed. However, in general, since the shape change in the longitudinal direction of the analysis chip 1 is not as great as the shape change in the thickness direction, it is usually preferable to form the partition wall 9b to prevent the shape change in the thickness direction.
[0110]
In addition, here, the force applied to the chip 1 as a force applied to the chip 1 has been described as an example, but the partition wall 9b can prevent shape deformation caused by the other force. For example, it is possible to prevent shape deformation caused by various forces applied to the chip 1 such as a force due to a pressure fluctuation of the liquid specimen Fs in the flow path 5 and a change in atmospheric pressure.
[0111]
Further, in the present embodiment, the chip 1 is configured by assembling the substrate 4, the plates 8 and 9, and the lid portion 2 with a holder, but the above-described substrate 4, the plates 8, 9, and the lid portion 2 are bonded together. If fixed, it is possible to prevent deformation when force is applied not only in the compression direction but also in the tension direction and the displacement direction.
[0112]
As described above, since the partition wall 9b, which is a support member, can prevent the chip 1 from being deformed, the flow velocity distribution of the liquid specimen Fb flowing through the flow path 5 and the light transmitted through the flow path 5 as in the past. Accurate analysis can be performed without changing the optical path length or the direction of the optical axis. Furthermore, as a result of changes in the type, flow rate, pressure, etc. of the sample liquid Fs over time, holding power of the analysis chip 1 over time, or changes in humidity, temperature, etc., holding of the analysis chip 1 Even when the shape deformation of the analysis chip 1 is accompanied by temporal fluctuations, such as characteristics such as force and viscosity of the liquid specimen Fs change with time, the deformation of the chip 1 due to temporal fluctuations. Can be prevented, and an accurate analysis can be performed.
[0113]
Further, as described above, since the pitch of the specific substance 61 can be further narrowed compared to the conventional case and the number of spots per unit area can be increased, the analysis can be efficiently performed with a small amount of the liquid specimen Fs. it can. That is, a small amount of the liquid specimen Fs can be realized. Further, conventionally, the liquid specimen Fs also circulates through the portion where the specific substance 61 of the flow path 5 is not fixed. However, in this embodiment, the specific substance 61 of the flow path 5 is not fixed. Since at least a part of the portion is occupied by the partition wall 9b, it is possible to further reduce the amount of the liquid specimen Fs by the volume that the partition wall 9b occupies.
[0114]
That is, a cross section obtained by cutting the flow path 5 along a plane orthogonal to the flow direction of the liquid specimen Fs in the flow path 5 without reducing the area of the portion where the specific substance 61 is fixed (hereinafter referred to as flow path cross section as appropriate). The contact area between the liquid specimen Fs and the specific substance 61 is made larger than before, and the amount of the liquid specimen Fs flowing through the flow path 5 is made smaller than before. As a result, even a small amount of the liquid specimen Fs can be brought into contact with the plurality of specific substances 61 by a single distribution to realize high-throughput analysis, and the analysis can be performed efficiently.
[0115]
Further, since the chip 1 is formed of a material having high strength, the deformation of the chip 1 can be prevented. Further, if the strength of the material of the chip 1 is increased, the flow path 5 is not damaged even if a high pressure is applied to the flow path 5, so that even if bubbles 201 are generated in the flow path 5, It is possible to forcibly discharge the bubbles 201 by applying a high pressure to the liquid specimen Fs flowing through the channel 5.
[0116]
Furthermore, since the frequency of redoing the analysis work caused by the generation of the bubbles 201 and the amount of the liquid sample Fs used therewith are reduced, there is an advantage that the analysis can be performed efficiently.
Further, according to the method for manufacturing the analysis chip 1, the plate 9 is fixed to the substrate 4 using a member having a lower affinity for the fluid specimen Fs than the substrate 4, and then the upper portion is opened. The specific substance-containing liquid is dropped through the slit-shaped holes 9a. At this time, since the fluid specimen Fs is guided to the substrate 4, the specific substance 61 can be stably fixed in the slit-like flow path 9a.
[0117]
As can be seen from the above, in the present specification, the convex member can divide the flow path 5 or is interposed in an opposing surface of the flow path 5 to support the opposing surface. It refers to a member that protrudes as much as possible.
[0118]
Further, in the present embodiment, the chip 1 is configured by sandwiching the plate 8 and the plate 9 with the substrate 4 and the lid portion 2, but as shown in FIGS. 7A to 7C, the collective flow path portion is formed. The holes 81 and 82 of the plate 8 may be formed in the lid portion 2. That is, in this case, grooves (recesses) 21 ′ and 22 ′ that form the collecting flow path portion having the same shape as the holes 81 and 82 are directly formed on the lower surface of the lid 2. Thereby, since it is only necessary to sandwich the plate 9 between the substrate 4 and the lid portion 2, the chip 1 can be easily manufactured. Further, as shown in FIGS. 8 (a) and 8 (b), grooves (recesses) 21 'and 22' for forming the collecting flow path portion having the same shape as the holes 81 and 82 are directly formed on the lower surface of the lid portion 2. In addition, the slit channel 4a may be formed directly on the substrate 4 by the slit groove without using the plate 9. Thereby, since it is only necessary to superimpose the substrate 4 and the lid 2, the chip 1 can be manufactured more easily. At this time, the partition wall 4b between the slit-like grooves 4a functions as a convex member and a support member.
[0119]
Further, as shown in FIGS. 9A to 9C, holes 81 and 82 of the plate 8 may be formed in the substrate 4 instead of using the plate 8. That is, in this case, grooves (recesses) 43 and 44 having the same shape as the holes 81 and 82 are formed directly on the upper surface of the substrate 4 and the lid 2 and the plate 9 are overlapped. In addition, holes 91 ′ and 92 ′ are formed in the plate 9 so that the injection port 21 and the hole 91 ′ and the discharge port 22 and the hole 92 ′ are aligned and the injection port 21 and the discharge port 22 communicate with the groove. . Thereby, since it is only necessary to sandwich the plate 9 between the substrate 4 and the lid portion 2, the chip 1 can be easily manufactured.
[0120]
Further, the holes 8 and 82 of the plate 8 may be formed in the plate 9 and the plate 9 may be sandwiched between the substrate 4 and the lid portion 2, and the chip 1 can also be manufactured by such a configuration (FIG. 33 (a)). In this case, as will be described later, the chip 1 can be easily manufactured by using screen printing, printing such as inkjet, or coating.
[0121]
Further, the internal flow path may be formed in a plurality of the lid portion 2, the intermediate plate 8, and the substrate 4. For example, as shown in FIGS. 45 (a) to 45 (c), a slit-like hole 83 connecting the upstream collecting flow path portion 81 and the downstream collecting flow path portion 82 is formed in the plate 8, and further, the substrate 4, the slit-shaped groove 4a may be formed, and both the slit-shaped hole 83 and the slit-shaped groove 4a may function as an internal flow path. Although only one slit-like hole 83 is formed in the plate 8 here, it goes without saying that a plurality of slit-like holes 83 may be formed.
7 to 9, 33 and 45, the same reference numerals as those used in FIGS.
[0122]
[2] Second embodiment
The analysis chip according to the second embodiment of the present invention is configured as an analysis chip (hereinafter, referred to as a sensor chip) used for an SPR sensor using surface plasmon resonance (SPR: Surface Plasmon Resonance).
[0123]
Hereinafter, the SPR sensor and the sensor chip will be described with reference to FIGS. 10 and 11.
10 and 11 show a second embodiment of the present invention. FIG. 10 is a schematic system configuration diagram of the SPR sensor, and FIG. 11 is a schematic exploded perspective view for explaining the configuration of the sensor chip. FIG. Note that members described in the first embodiment are denoted by the same reference numerals and description thereof is omitted.
[0124]
As shown in FIG. 10, the SPR sensor includes a sensor chip 1A, a light source 100 for irradiating light to the sensor chip 1A, and a detector for detecting reflected light from the sensor chip 1A (here, a CCD (Charge Coupled). Device) camera] 101. In FIG. 10, the optical axes of incident light from the light source 100 and reflected light from the sensor chip 1A are shown in a direction perpendicular to the flow direction, but the directions of the optical axes of the incident light and reflected light are limited to this. For example, the optical axis of the incident light may be parallel to the flow direction, and the optical axis of the reflected light may be changed from the direction of the optical axis of the incident light by being reflected by the sensor chip 1A. . Furthermore, light may be irradiated from the back surface (substrate 4 side) of the sensor chip 1 and the reflected light may be observed and analyzed on the back surface of the sensor chip 1 (substrate 4 side). The substrate 4 must be formed of a material that can transmit incident light and reflected light.
[0125]
Similar to the analysis chip 1 of the first embodiment described above (see FIG. 1), the sensor chip 1A includes a slit-like flow path 9a and collecting flow path portions 81 and 82, and a liquid feed pump (not shown). ) Allows the liquid specimen Fs to circulate through the slit-like flow passage 9a and the collecting flow passage portions 81 and 82.
[0126]
As shown in FIG. 11, the sensor chip 1A is different from the analysis chip 1 of the first embodiment in the configuration of the substrate 4, and the lid 2 and the plates 8, 9 are made of a particularly transparent material. Yes.
A metal layer 41 is coated on one surface of the substrate 4 facing the slit-shaped flow path 9a when the chip 1 is assembled. As in the first embodiment, after the plate 9 is overlaid on the substrate 4, a diffraction grating 42 is formed on the surface coated with the metal layer 41 as an optical structure for generating an evanescent wave, and the reaction is performed. Part 6 (a plurality of specific substances 61) is formed. The reaction unit 6 is directly fixed to the metal layer 41 or is fixed to an organic film formed on the metal layer 41.
[0127]
When light is emitted from the light source 100 to the substrate 4 through the transparent lid 2 and the plates 8 and 9, surface plasmon waves generated on the surface of the metal layer 41 by this light are caused by the diffraction grating 42. Resonating with the evanescent wave induced in the metal layer 41, the energy of the light component having a specific incident angle or a specific wavelength among the light irradiated to the metal layer 41 is absorbed by the metal layer 41.
Therefore, the reflected light from the metal layer 41 has a weak energy of a light component having a specific incident angle or a specific wavelength.
[0128]
The angle and wavelength of the evanescent wave generated on the metal layer 41 vary according to the amount of the detection species captured by the metal layer 41 or the specific substance 61 fixed to the organic film formed on the metal layer 41, In accordance with this, the angle and wavelength of the reflected light to be absorbed change. The organic film here includes a known structure. Further, as the function of the organic film, it is desirable that the specific substance 61 is stably immobilized on the metal layer 41 to suppress nonspecific adsorption. For example, the functional group for binding to the biological material includes any one of amino, aldehyde, epoxy, carboxyl, carbonyl, hydrazide, hydroxyl, vinyl group, and isothiocyanate, isonitrile, xanthate, It contains a straight chain polymer containing any of diselenide, sulfide, selenide, selenol, thiol, thiocarbamate, nitrile, nitro, phosphine, or a hydrocarbon chain containing double or triple bonds. Further, a hydrogel (agarose, alginic acid, carrageenan, cellulose, dextran, polyacrylamide, polyethylene glycol, polyvinyl alcohol, etc.) may be used as a matrix. Alternatively, a structure using an LB film, a self-assembled monomolecular film, a lipid bilayer, or the like may be used.
[0129]
Therefore, the light intensity of the reflected light from each specific substance 61 in the reaction unit 6 is monitored by the CCD camera 101, and the concentration of the detection species in the test fluid is measured in real time by detecting such changes in angle and wavelength. it can.
The material of the metal layer 41 is not limited as long as it can induce surface plasmon waves, and is, for example, gold, silver, aluminum or the like.
[0130]
Further, the diffraction grating 42 can be embodied on the surface of the metal layer 41 by forming irregularities on the surface of the substrate 4 and forming the metal layer 41 by thinly laminating metal thereon by sputtering or the like.
The unevenness formed to provide the diffraction grating 42 on the substrate 4 is formed by, for example, cutting the substrate 4 and may be mechanically performed as a cutting method or chemically performed by an etching technique or the like. It may be a thing.
[0131]
Furthermore, when the substrate 4 is made of a resin material, it is possible to press the stamper on which the unevenness is formed by, for example, photolithography to form the unevenness before the resin material is completely solidified, The uneven shape may be transferred from the stamper by injection molding.
[0132]
Since the analysis chip (sensor chip) 1A according to the second embodiment of the present invention is configured as described above, the bubbles generated by the entrainment of the liquid specimen Fs are the same as the analysis chip 1 according to the first embodiment. Can be suppressed, the analysis chip 1A can be prevented from being deformed, and the amount of the liquid sample Fs can be reduced.
[0133]
A major feature of the analysis chip used in the SPR sensor is that the state of interaction in the reaction unit 6 (a plurality of specific substances 61) is detected optically and online.
For example, if bubbles remain in the reaction part (that is, the measurement region) 6, not only the interaction between the specific substance 61 and the detection species is inhibited, but the above optical measurement cannot be performed. However, according to the present analysis chip 1A, since the generation of bubbles can be suppressed, there is an advantage that online analysis by optical measurement as described above can be performed stably.
[0134]
In the analysis using such SPR, not only the same liquid specimen Fs is circulated through the microchannel chip, but also the plurality of liquid specimens Fs are continuously circulated with a buffer interposed therebetween. It is also possible to analyze a series of binding-dissociation between the measurement object of these liquid specimens Fs and the specific substance.
[0135]
In this embodiment, a CCD camera is used as the detector 101. However, the detector 101 is not limited to this, and any detector such as a photodiode, a photomultiplier tube, or photosensitive paper may be used. it can.
[0136]
[3] Third embodiment
12 to 14 show the structure of an analysis chip as a third embodiment of the present invention. FIG. 12 (a) is a schematic assembly perspective view, and FIG. 12 (b) is a schematic disassembly. FIG. 13 is a schematic bottom view of the plate, FIG. 14A is a sectional view taken along the line Y1-Y1 in FIG. 12A, and FIG. 14B is a sectional view taken along the line X3-X3 in FIG. It is. Note that members described in the first embodiment are denoted by the same reference numerals and description thereof is omitted.
[0137]
As shown in FIGS. 12A and 12B, the analysis chip (also simply referred to as a chip) 1 </ b> B includes a lid portion 2, a plate 10, and a substrate 4.
The present analysis chip 1 is characterized in that a plate (intermediate plate) 10 is provided instead of the plates 8 and 9 in the first embodiment. Therefore, in particular, the plate 10 will be described in detail below.
Pipes 26 and 27 (see FIG. 14) are inserted into the inlet 21 and the outlet 22 of the lid 2, respectively, so that it can be easily connected to an external liquid feed pump or a tube connected to a waste liquid tank. It has become so.
[0138]
As shown in FIGS. 12A and 12B, holes 11 and 12 communicating with the inlet 21 and the outlet 22 of the lid 2 are formed in the plate 10.
Further, as shown in FIG. 13, a concave portion 13 formed on the lower surface of the plate 10 (side facing the substrate 4) so as to become wider from the hole 11 toward the intermediate portion in the flow direction of the plate 10, A concave portion 14 is provided which is formed so as to become narrower from the intermediate portion in the flow direction of the plate 10 toward the hole 12.
[0139]
Note that, when the plate 10 is overlaid on the substrate 4, the recesses 13 and 14 of the plate 10 are closed at their openings to form a flow path in which the liquid specimen Fs collects. Therefore, the space formed by the substrate 1 and the recesses 13 and 14 of the plate 10 is also referred to as the collecting flow path portions 13 and 14.
In addition, a plurality of slit-like grooves 10a that are divided and formed in the width direction by the partition wall 10b are formed in the intermediate portion in the flow direction of the plate 10 (the region between the recess 13 and the recess 14). Therefore, by overlapping the plate 10 on the substrate 4, the concave portions 13 and 14 and the intermediate portion of the plate 10 form the flow path 5 through which the liquid specimen Fs flows.
[0140]
In addition, since the above-described slit-shaped groove 10a is closed by opening the plate 10 with the substrate 4 to form a slit-shaped internal flow path (slit-shaped flow path), the slit-shaped groove is hereinafter referred to as a slit-shaped groove. The groove and the slit-like channel are denoted by the same reference numeral 10a.
At this time, the aspect ratio (vertical dimension / horizontal dimension) of the cross section of the slit-shaped flow path 10a is about 0.005 (for example, 5 μm in length, 1 mm in width) to 100 (for example, 1000 μm in length, 10 μm in width). It is preferable that the slit-shaped channel 10a is formed so as to be within the range. In general, the slit-shaped channel 10a is 5 mm.²It may be formed to have the following cross-sectional area, preferably 100 μm²More than 5mm²Or less, more preferably 2000 μm²0.3 mm or more²It is good to form so that it may have the following cross-sectional areas. By configuring the slit-like channel 10a in this way, the generation of bubbles due to the wraparound of the liquid specimen Fs in the channel can be more reliably suppressed.
[0141]
In addition, as a method of forming such recesses 13 and 14 and slit-like groove 10a, transfer techniques represented by machining, injection molding and compression molding, dry etching (RIE, IE, IBE, plasma etching, laser etching, Laser ablation, blasting, electrical discharge machining, LIGA, electron beam etching, FAB), wet etching (chemical erosion), monolithic molding such as stereolithography and ceramic laying, etc., coating various materials in layers, vapor deposition, sputtering, deposition Surface micro-machining for forming a fine structure by removing partly, and a method for forming a flow path component by dropping with an ink jet or a dispenser (that is, the recesses 13 and 14 and the intermediate portion in the flow direction are integrally formed as a recess) And then in the middle part Re dropwise flow duct forming material along a direction, a method of forming a partition wall 10b), may be selected and used stereolithography and the like as appropriate.
[0142]
Next, a method for manufacturing the analysis chip 1B will be described. First, a specific substance-containing liquid is dropped onto a target position of the substrate 4 by an injector or the like that can be positioned, and specified at a reference interval on the substrate 4. The substance 61 is fixed.
After that, as shown in FIG. 14B, the plate 10 is positioned and placed on the substrate 4 so that the partition walls 10b of the plate 10 are located between the rows of the specific substance 61. Set the lid part 2 to. In addition, you may make it set to the board | substrate 4 what joined the lid | cover part 2 and the plate 10 previously.
[0143]
Since the analysis chip 1B as the third embodiment of the present invention is configured as described above, the liquid specimen Fs injected into the injection port 21 of the lid 2 is as shown in FIG. , And flows through the collecting channel portion 13 through the holes 11 of the plate 10. Then, it flows through each slit-shaped channel 10 a and contacts the specific substance 61.
Thereafter, the liquid specimen Fs gathers from the slit-like flow passages 10 a to the collecting flow passage portion 14 and is discharged out of the chip 1 through the holes 12 of the plate 10 and the discharge ports 22 of the lid portion 2.
[0144]
As described above, according to the present analysis chip 1B, the collecting flow path portion 13 becomes wider from the upstream end portion toward the intermediate portion in the flow direction, so that the liquid specimen Fs can be smoothly transferred to the intermediate portion in the flow direction. I can guide you. In addition, since the collecting flow path portion 14 becomes narrower as it goes from the intermediate portion in the flow direction to the downstream end portion, the liquid specimen Fs can be smoothly guided to the downstream end portion.
[0145]
Further, by providing the partition wall 10b in the flow path forming the sheet-shaped space to form a narrower flow path (slit-shaped flow path) 10a, the generation of bubbles due to the entrainment of the liquid specimen Fs is suppressed. can do.
Therefore, according to the analysis chip 1B, similarly to the effect of the first embodiment, adverse effects due to the retention of bubbles (inhibition of the flow of the liquid specimen Fs, inhibition of contact between the specific substance 61 and the liquid specimen Fs, liquid Fs) And non-uniformity in the temperature of the measurement system due to the difference in the heat transfer coefficient between the bubble 201 and the bubble 201 and the obstruction of measurement due to the bubble 201 remaining in the optical path), and the reliability of the analysis can be improved. There is. Furthermore, there is an advantage that the work of removing bubbles is not required and the analysis work can be performed efficiently.
[0146]
Further, when assembling the chip 1B using a holder, pressure is applied to the chip 1B. As in the first embodiment, a plurality of partition walls 10b formed across the width direction of the chip 1 are column members. Therefore, the pressure resistance of the chip 1B can be improved, and the shape change in the thickness direction of the chip 1B can be prevented. Thereby, non-uniformity in the flow velocity distribution due to the deflection of the chip 1B can be prevented, and in the analysis using light irradiation, variations in the optical path length and changes in the optical axis can be prevented, thereby improving the accuracy of the analysis result. be able to.
Furthermore, as in the first embodiment, it is possible to reduce the amount of the liquid specimen Fs.
[0147]
In the present embodiment, the chip 1B is configured with the plate 10 sandwiched between the substrate 4 and the lid 2. However, as shown in FIGS. 15A and 15B, the concave portions 13 and 14 of the plate 10 and the slit shape are formed. The recesses 21 ′ and 22 ′ that form the collecting flow path portions and the slit-shaped grooves 2 a that form the internal flow paths corresponding to the grooves 10 a may be directly formed on the lower surface of the lid portion 2. In this case, the substrate 1 and the lid 2 need only be overlapped without providing the plate 10, so that the chip 1B can be easily manufactured. Reference numeral 2b denotes a partition wall.
The analysis chip 1 manufactured in this way is disposed so as to face the substrate 4 and the substrate 4, and forms a sheet-like space having the flow path 5 between the substrate 4 in cooperation with the substrate 4. The flow path 5 includes a partition wall 10b having both functions of a column member and a partition member.
[0148]
Further, as shown in FIGS. 16 (a) and 16 (b), the concave portions 43 and 44 that form the collecting flow path portions and the internal flow paths corresponding to the concave portions 13 and 14 and the slit-like grooves 10a of the plate 10 are provided. The slit-like groove 4a to be formed may be formed directly on the upper surface of the substrate 4. In this case, the substrate 1 and the lid 2 need only be overlapped without providing the plate 10, so that the chip 1B can be easily manufactured. 4b represents a partition wall.
Further, for example, when the chip 1B is manufactured by using only the substrate 4 and the lid part 2 as described above, as shown in FIG. It may be formed on the upstream and downstream side surfaces, or on the lower surface of the substrate 4 as shown in FIG.
[0149]
Further, in the present embodiment, as indicated by the symbol R in FIGS. 13, 15A, and 16B, the distances from the holes 11 or 12 to the end portions of the partition walls 10b are substantially equal. The partition walls 10b, 2b, and 4b are provided, but as indicated by reference numeral R1 in FIG. 18, the partition wall 10b that is closer to the center in the width direction is made longer (the hole 11 or the hole 12 and the partition wall). The distance from the end portion of 10b may be shorter than those in FIGS. 13, 15 (a), and 16 (b). Needless to say, the example shown in FIG. 18 can be similarly applied to the examples shown in FIGS. 15 (a) and 16 (b).
[0150]
[4] Fourth embodiment
FIG. 19A is a bottom view of a plate (intermediate plate) for explaining the fourth embodiment of the present invention.
As shown in FIG. 19A, the analysis chip 1C as the fourth embodiment of the present invention is formed so that the partition wall 10b of the intermediate plate 10 extends from the hole 11 serving as the inlet to the hole 12 serving as the outlet. Other than that, the configuration is the same as that of the third embodiment described above with reference to FIGS. Note that members described in the third embodiment are denoted by the same reference numerals and description thereof is omitted.
[0151]
That is, in the present embodiment, the partition wall 10b is formed not only in the middle part in the flow direction but also in the recesses 13 and 14, and the inlets of the respective internal flow paths 10a divided by the partition wall 10 are opened in the holes 11, The outlet opens into the hole 12. Therefore, in this embodiment, the recessed parts 13 and 14 do not constitute a collecting flow path part, but constitute a part of the internal flow path 10a.
[0152]
Since the fourth embodiment of the present invention is configured as described above, the same actions and effects as those of the third embodiment can be obtained except for the actions and effects of the collecting flow path portion. In the third embodiment, since the internal flow path can be formed up to the concave portion that has formed the collecting flow path portion, it is possible to reliably prevent the generation of bubbles in a wider portion in the flow path 5, In addition, it is possible to prevent the chip from being deformed and to further promote the reduction in the amount of the liquid specimen Fs. As a result, analysis can be performed more accurately, and a wider reaction section can be provided, enabling efficient analysis.
[0153]
Moreover, it is preferable that the distance of each internal flow path 10a becomes equal. Further, depending on the pressure of the liquid specimen Fs, the shape of the partition wall 10b, the characteristics of the wall surface of the flow path 5 and the like, even if bubbles are generated, the bubbles can be easily removed by the back pressure.
[0154]
Also in this embodiment, as in the third embodiment, as shown in FIG. 19B, groove portions (recess portions) 23, 24 and slit grooves corresponding to the recess portions 13, 14 and the slit groove 10a of the plate 10 are provided. 2a may be formed directly on the lower surface of the lid portion 2, and as shown in FIG. 19C, the groove portions (recess portions) 46, 47 corresponding to the recess portions 13, 14 of the plate 10 and the slit-like groove 10a. The slit-shaped grooves 4a may be directly formed on the upper surface of the substrate 4.
The configuration shown in FIG. 19B is the same as that shown in FIGS. 15A and 15B, and the configuration shown in FIG. The configuration is the same as that shown in FIGS. 16 (a) and 16 (b).
[0155]
[5] Fifth embodiment
FIG. 20A is a bottom view of a plate (intermediate plate) for explaining the fifth embodiment of the present invention.
The analysis chip 1D as the fifth embodiment of the present invention has a basic configuration in common with the above-described third embodiment as shown in FIG. That is, the configuration shown in FIGS. 20A and 20B is the same as the third embodiment except for the other configurations. Note that members described in the third embodiment are denoted by the same reference numerals and description thereof is omitted. Furthermore, as shown in FIG. 20B, the present embodiment is characterized in that a contracted portion 10c is formed at the downstream end of the internal flow path 10a.
[0156]
FIG. 20B is an enlarged view of the XXb portion of FIG. As shown in FIG. 20B, the contracted portion 10c is a portion formed so that the internal flow path 10a is gradually narrowed, that is, the cross-sectional area perpendicular to the flow direction of the internal flow path 10a is reduced. Yes, here, it refers to a portion formed so that the width of the internal flow path 10a becomes gradually narrower.
[0157]
Since 5th Embodiment of this invention is comprised as mentioned above, in addition to the effect similar to 3rd Embodiment, it can suppress that a bubble remains in the downstream end part of the internal flow path 10a.
More specifically, it has been empirically known that bubbles 201 are easily formed at the downstream end of the internal flow path 10a as shown in FIG. This is because the liquid sample Fs moves from the internal flow channel 10a to the downstream collective flow channel portion 14 when the downstream end of the internal flow channel 10a has a shape as shown in FIG. It is considered that the back pressure suddenly disappeared at the downstream end of the internal flow path 10a and the bubbles 201 were retained. 20C shows an example in which the downstream end portion of the internal flow path 10a is formed so as to gradually expand in the width direction (that is, the thickness of the partition wall 10b is gradually reduced), but the internal flow path 10a is shown. Even if the downstream end portion is formed to be constant in the width direction, the bubbles 201 may be formed in the same manner. On the other hand, in the present embodiment, the above-described constricted part 10c is formed, and when the liquid specimen Fs moves from the internal flow path 10a to the downstream collecting flow path part 14, as described above, abruptly, In order to prevent the back pressure from being applied, the linear velocity of the liquid specimen Fs is increased by the contracted portion 10c shown in FIG. 20B, thereby suppressing the bubbles 201 from remaining in the internal flow path 10a. Can do.
[0158]
Needless to say, the configuration of the present embodiment can be applied to other embodiments or modifications thereof.
[0159]
[6] Sixth embodiment
FIG. 21A is a schematic enlarged view of a section obtained by cutting a portion where a partition wall of a flow path is formed by a plane orthogonal to the width direction of the flow path in order to explain a sixth embodiment of the present invention. FIG.
As shown in FIG. 21A, the analysis chip 1E as the sixth embodiment of the present invention has the same basic configuration as that of the third embodiment described above. Note that members described in the third embodiment are denoted by the same reference numerals and description thereof is omitted. In this embodiment, as shown in FIG. 21 (a) and FIG. 21 (b) showing the XXIb portion in FIG. 21 (a) in an enlarged manner, the partition wall 10b is partitioned on the surface facing the substrate 4. A layer 10 t of Teflon (registered trademark) is formed as a material layer (adhesion reduction layer) for reducing the adhesion between the wall 10 b and the substrate 4, thereby reducing the adhesion between the partition wall 10 b and the substrate 4. The partition wall 10b and the substrate 4 are separated by a very small distance.
[0160]
As a degree of reducing the adhesion, as a result of the reduction in the adhesion between the partition wall 10 b and the substrate 4, the liquid specimen Fs, which is a fluid, enters between the partition wall 10 b and the substrate 4, and the entered liquid The partition wall 10c can support the substrate 4 through the thin liquid layer of the specimen Fs, and the specific adhesion varies depending on the type of the liquid specimen Fs, the conditions at the time of analysis, and the like.
Further, the end portion of the partition wall 10 b on the plate 10 side is formed integrally with the plate 10.
[0161]
Since 6th Embodiment of this invention is comprised as mentioned above, in addition to the effect similar to 3rd Embodiment, the shape deformation by the stress between the partition wall 10b and the board | substrate 4 can be prevented.
Details will be described below. When the partition wall 10b and the substrate 4 are bonded or highly adhered, if the excessive stress acts on the contact portion between the partition wall 10b and the substrate 4, the analysis chip 1E can be deformed by the stress. There is sex. That is, an ideal shape that occurs when the partition wall 10b and the substrate 4 are bonded to each other when the analysis chip 1E is manufactured, or when the liquid specimen Fs is flowed after bonding or when the temperature changes (ie, when the temperature changes). The shape of the analysis chip 1E may be deformed by a stress resulting from deviation from the shape when no force such as stress or pressure is applied.
[0162]
Therefore, in the present embodiment, the adhesion between the partition wall 10b and the substrate 4 is reduced, and when the liquid specimen Fs, which is a liquid, flows through the flow path 5, it is partially between the partition wall 10b and the substrate 4. The liquid specimen Fs is allowed to enter, and the partition wall 10b can support the substrate 4 through a thin liquid layer of the liquid specimen Fs that has entered, and thus the plate 10 is supported by the substrate 4. Thereby, it is possible to prevent excessive stress from being transmitted between the partition wall 10b and the substrate 4, so that stress is generated between the partition wall 10b and the substrate 4 or stress is transmitted. Shape deformation can be prevented.
[0163]
Further, the degree of the distance between the partition wall 10b and the substrate 4 is so small that the partition wall 10b can support the substrate 4 via the liquid specimen 10b existing as a thin liquid layer state, and is normally expected. It is preferable to set it small enough to suppress the deformation of the analysis chip 1E.
[0164]
The adhesion reducing layer may be a layer other than a Teflon (registered trademark) layer, and may be any layer that can reduce the adhesion between the surface of the partition wall 10 b and the surface of the substrate 4.
[0165]
Further, the partition wall 10b may be supported by the substrate 4 via the liquid specimen Fs between the partition wall 10b and the substrate 4 by other methods. For example, by the wet etching such as chemical erosion or the dry etching such as reactive ion etching (Deep Reactive Ion Etching), the height of the partition wall 10b (that is, the flow path) by a very small distance of about several nm to several tens of nm. The distance in the height direction) may be formed low.
[0166]
However, in the present embodiment, unlike the third embodiment, the liquid specimen Fs may leak between the adjacent internal flow paths, that is, the slit-shaped grooves 10b, between the partition wall 10b and the substrate 4. Therefore, it is preferable to use the technique described in this embodiment when such a leak may occur.
[0167]
In the present embodiment, attention is paid to a portion between the partition wall 10b and the substrate 4. However, when the analysis chip 1E has another structure, a liquid is formed according to the structure of the analysis chip 1E. A portion supported by the specimen Fs may be selected. Furthermore, when the flow path 5 is formed facing the lid portion 2, the space between the partition wall 10b and the surface of the lid portion 2 on the flow path 5 side is supported via the liquid specimen Fs. That's fine. The technique of the present embodiment can be applied to any part of the surface of the flow path 5 where stress is generated.
[0168]
Further, in the present embodiment, the case where the liquid specimen Fs is caused to flow between the partition wall 10b and the substrate 4 is described as an example. However, the fluid that enters between the partition wall 10c and the substrate 4 is arbitrary, for example, When analysis is performed while flowing a buffer other than the liquid specimen Fs, the partition wall 10b may support the substrate 4 through the buffer flowing in the flow path 5. Further, when a gas such as air or a mixture of gas and liquid stays between the partition wall 10b and the substrate 4, the partition wall 10b can support the substrate 4 through them. Good. However, if a gas stays between the partition wall 10b and the substrate 4, the gas continuously stays between the partition wall 10b and the substrate 4 throughout the analysis, and so on. It is preferable that the accuracy of analysis does not decrease due to gas.
[0169]
Furthermore, the fluid does not always have to exist between the partition wall 10b and the substrate 4. For example, even if the partition wall 10b and the substrate 4 directly contact each other at a certain moment during analysis, Therefore, it is sufficient that excessive stress does not occur between the partition wall 10b and the substrate 4.
[0170]
[7] Seventh embodiment
FIG. 22 is a schematic bottom view of a plate (intermediate plate) for explaining the seventh embodiment of the present invention, and FIGS. 23 (a) and 23 (b) are enlarged views for explaining the function thereof.
The basic configuration of the analysis chip 1F as the seventh embodiment of the present invention is common to the above-described third embodiment. That is, the configuration shown in FIGS. 22, 23 (a), and (b) is the same as that of the third embodiment except for the other configurations. Note that members described in the third embodiment are denoted by the same reference numerals and description thereof is omitted. In this embodiment, as shown in FIG. 22, the upstream and downstream ends of the partition wall 10b are formed in a line in the width direction, and the intermediate plate 10 side surface of the flow path 5 is processed to be hydrophilic. And a strip-like shape extending over the entire width of the flow channel 5 in the direction orthogonal to the flow direction of the flow channel 5 on the surface on the intermediate plate 10 side of the collective flow channel portion 13 on the upstream side of the flow channel 5 A plurality of hydrophobic portions (hydrophobic portions, hereinafter referred to as hydrophobic portions as appropriate) 5x are intermittently formed.
[0171]
Accordingly, the collecting flow path portion 13 is formed with a strip-like portion (hydrophilic portion, hereinafter referred to as a hydrophilic portion) 5y in which the surface of the intermediate plate 10 processed to be hydrophilic is exposed as a first affinity portion, Further, the hydrophobic portion 5x is formed as the second affinity portion. As a result, the hydrophilic portion 5y and the hydrophobic portion 5x are alternately arranged in the flow direction in the collective flow path portion 13. Here, the affinity part refers to a part having a certain affinity for a target substance. Therefore, the affinity part does not necessarily have a high affinity for the target substance.
[0172]
The hydrophobic portions 5x have the same thickness (the flow direction dimension of the flow path 5), and the hydrophilic portion 5y sandwiched between the hydrophobic portions 5x has the same thickness as the hydrophobic portion 5x. Has been. Although the said thickness is suitably set according to various conditions and it is not limited to being the same dimension, Generally it is 10 micrometers-1000 mm or less.
[0173]
Accordingly, as shown in FIG. 23A, when a part of the hydrophilic liquid specimen Fs arrives at the hydrophobic part 5x in advance, the hydrophobic part 5x is difficult to circulate, so that the liquid specimen Fs is separated from the hydrophobic part 5x. The subsequent liquid specimen Fs is in a state where the brake is not applied and is easily flowable, that is, the hydrophilic part 5y side where the liquid specimen Fs has not reached yet. As shown by F3, the flow starts.
[0174]
As a result, the liquid specimen Fs is prevented from moving partly over the hydrophobic part 5x until it reaches the hydrophobic part 5x over the entire width of the flow path 5. That is, the tip (gas-liquid interface) S of the liquid specimen Fs can be aligned along the width direction B as shown in FIG.
Here, as described above, the hydrophobic portion 5x is provided only in the intermediate plate 10. However, the hydrophobic portion 5x may be formed on the surface of the flow path 5 on the substrate 4 side as long as it is the surface of the flow path 5. It can be provided at any place. The hydrophobic portion 5x is most preferably formed over the entire circumference of the cross section of the flow path 5. However, since the flow path 5 has a very small diameter, the resistance received from the wall surface of the flow path 5 is large. Even if only the hydrophobic portion 5x is provided in the intermediate plate 10 as in the embodiment, the flow of the liquid specimen Fs can be suppressed and the tip position of the liquid specimen Fs can be aligned as described above.
[0175]
Since the analysis chip according to the seventh embodiment of the present invention is configured as described above, in addition to the same effects as those of the third embodiment, bubbles due to wraparound of the liquid specimen Fs by the hydrophilic portion 5y and the hydrophobic portion 5x. Can be prevented more reliably, and an accurate and efficient analysis can be performed.
[0176]
Further, since the hydrophilic portion 5y and the hydrophobic portion 5x are formed in the collective flow path portion 13 upstream of the reaction portion 6 in the flow direction, bubbles are generated before the liquid specimen Fs arrives at the reaction portion 6. Therefore, accurate analysis can be performed.
[0177]
In this embodiment, the first affinity part 5y is made hydrophilic and the second affinity part 5x is made hydrophobic. However, for example, the surface of the flow path 5 can be formed without forming both the hydrophilic part and the hydrophobic part. Is partially hydrophobized, or on the contrary, only a portion excluding a predetermined portion (a portion corresponding to the hydrophobic portion 5x) on the surface of the flow path 5 is hydrophilized, so that the second portion is more than the first affinity portion. The affinity part may be configured to have a lower affinity for the liquid fluid Fs (here, the degree of hydrophilicity). However, when both the hydrophilic part 5y and the hydrophobic part 5x are provided as in this embodiment, the generation of bubbles can be more effectively suppressed.
[0178]
Any method may be used for hydrophilization and hydrophobization. Specific examples include acrylic resin, polycarbonate, polystyrene, silicon, polyurethane, polyolefin, polytetrafluoroethylene, When using hydrophobic materials with relatively low affinity, such as polypropylene, polyethylene, and thermoplastic elastomers, surface coating, wet chemical modification, gas modification, surfactant treatment, corona, etc. Discharge, roughening, metal vapor deposition, metal sputtering, UV treatment, methods involving application of hydrophilic functional groups or hydrophilic molecules depending on the processing atmosphere (plasma method, ion implantation method, laser treatment, etc.) Is mentioned.
[0179]
Further, when a hydrophilic material having a relatively high affinity such as glass, metal, or ceramic is used for the surface of the flow path 5, as a method for partially hydrophobizing this, a hydrophobic substance such as an adhesive or wax is used. Examples thereof include surface coating, surface grafting, and methods (plasma method, ion implantation method, laser treatment, etc.) involving application of hydrophobic functional groups or hydrophobic molecules depending on the processing atmosphere to the surface.
[0180]
When the modification (hydrophilization or hydrophobization) of the wall surface of the flow path 5 is performed as described above, the above-described treatment may be performed only on the portion of the wall surface of the flow path 5 that is to be modified, It is also possible to perform the above-described process collectively on the entire wall surface of the flow path 5 by masking.
It is also possible to form a partial pattern by assembling different types of hydrophilic and hydrophobic materials. That is, for example, a hydrophobic material may be formed on the wall surface of the hydrophilic flow path 5 to form a hydrophobic portion.
[0181]
Further, in the present embodiment, the strip-like hydrophilic portion 5y and the hydrophobic portion 5x are formed over the entire width of the flow path 5, but the width dimension, the number thereof arranged in the flow direction, the mutual interval, and the shape thereof are as follows. Is optional.
[0182]
Further, the strip-like hydrophilic portion 5y and the hydrophobic portion 5x do not necessarily have to be formed in a strip shape orthogonal to the flow direction of the flow path 5. For example, as shown in FIG. You may form in the strip | belt shape extended so that it may cross | intersect. Further, the hydrophobic portion 5x may not be formed completely over the entire width of the flow path 5. The extending angle and length of the hydrophilic portion 5y and the hydrophobic portion 5x, that is, the first and second affinity portions may be appropriately changed according to the configuration of the analysis chip 1F.
[0183]
Further, the hydrophilic portion 5y and the hydrophobic portion 5x may have a non-linear shape. For example, as shown in FIG. 25, when the distances from the holes 11 and 12 to the end of the partition wall 10b are formed to be substantially equal, the hydrophobic part 5x is formed in an arc shape whose distance from the hole 11 is substantially equal. Is preferred.
In the present embodiment, a plurality of first and second affinity portions are formed, but it is sufficient that at least one of them is provided. However, of course, it is preferable to arrange a plurality along the flow direction. In other words, if there is a large variation in the tip of the liquid specimen Fs when it reaches the hydrophobic portion 5x, it is difficult to sufficiently suppress this. Therefore, by arranging a plurality of hydrophobic portions 5x in the flow direction, the tip of the liquid specimen Fs is repeatedly arranged. By aligning them, it will be possible to prevent the air from being caught.
[0184]
Further, the first affinity part 5y and the second affinity part 5x may be provided anywhere in the flow path 5, for example, as shown in FIG. 26 (a), the first affinity part 5y and the second affinity part 5x. The part 5x is provided in the middle part of the flow path 5, and as shown in FIG. 26B, the first affinity part 5y and the second affinity part 5x are provided in the downstream collecting flow path part 14, As shown in 26 (c), the first affinity portion 5 y and the second affinity portion 5 x may be provided alternately over the entire length of the flow path 5.
The configurations shown in FIGS. 24, 25, and 26 (a) to 26 (c) are the same as those in the third embodiment.
[0185]
In the above-described embodiment, an example in which the liquid specimen Fs is hydrophilic has been described. However, the liquid specimen Fs may be hydrophobic, and in this case, the hydrophobic part is defined as a lipophilic part (relatively lipophilic part). The hydrophilic portion may be replaced with a lipophilic portion having higher lipophilicity than the oleophobic portion.
[0186]
In the above embodiment, the first affinity part 5y is composed of a hydrophilic part and the second affinity part 5x is composed of a hydrophobic part. However, the first affinity part 5x is formed as a rough surface part, and the second affinity part is formed. You may form 5y as a smooth surface part. Here, the smooth surface portion refers to a portion whose surface is smoother than the rough surface portion. Normally, when attention is paid to the paintability of the surface of the flow path 5, since the smooth surface has a lower affinity for the liquid specimen Fs than the rough surface, as in the above-described embodiment, bubbles are retained. Can be prevented. In the first embodiment, it is described that the pressure loss of the liquid specimen Fs flowing through the flow path 5 increases when the surface of the flow path 5 is rough. However, the pressure loss and the solid-gas-liquid three-phase boundary line are used. This is a concept different from the paintability (characteristic when the liquid specimen Fs is circulated for the first time in the flow path 5 where the liquid specimen Fs is not circulated), and the above-described content does not contradict the content of the first embodiment. Absent.
[0187]
In the above-described embodiment, an example in which only the first affinity part 5y and the second affinity part 5x are formed in the flow path 5 has been described. However, the present invention at least includes the first affinity part 5y and the second affinity part 5x. It is only necessary to have the affinity part 5x, and a new affinity part having a different affinity for the liquid specimen Fs from the first and second affinity parts may be further provided.
[0188]
[8] Eighth embodiment
FIG. 27 is a schematic perspective view for explaining an eighth embodiment of the present invention.
As shown in FIG. 27, the analysis chip unit 1G according to the eighth embodiment of the present invention includes a unit base 300 formed in a regular octagonal column and a rotation support shaft 302 that passes through the centers of both end faces of the regular octagonal prism. It is attached. Further, on the side surface of the regular octagonal prism of the unit base 300, unit chips 301 having the same configuration as the analysis chips of the first to third embodiments are held. Further, the unit base 300 is rotatable about the support shaft 301 as a rotation center. Since the structure of the unit chip 301 described in the first embodiment has been described in the first, third to seventh embodiments, description thereof will be omitted. Further, in FIG. 27, the flow path 5 of the unit chip 301 is seen through for the sake of explanation. That is, the lid 2 and the plates 8, 9, 10 actually cover the flow path 5.
[0189]
Since the analysis chip unit 1G as the eighth embodiment of the present invention is configured as described above, in use, analysis is performed using the unit chip 301 located at a predetermined position of the analysis chip unit 1G. That is, the liquid sample Fs is injected into the flow channel 5 from the injection port 21 of the unit chip 301 at a predetermined position, and the liquid sample Fs flowing in the flow channel 5 is analyzed. After the analysis, the liquid sample Fs is discharged from the discharge port 22. To the outside of the unit chip 301. When the analysis using the unit chip 301 is completed, the unit base 300 is rotated by a predetermined angle (for example, an integral multiple of 45 °) about the rotation support shaft 302, and another unit chip 301 is moved to a predetermined position. The analysis is performed using the unit chip 301 that has been moved to a predetermined position. This is repeated and the analysis using the analysis chip unit 1G is performed.
[0190]
If different types of specific substances 61 are fixed to each unit chip 301, simply by rotating the analysis chip unit 1G, it is possible to easily detect different types of predetermined substances, and efficiently analyze the liquid specimen Fs. Can be performed.
[0191]
In the present embodiment, the unit base 300 has a regular octagonal prism shape, but the shape of the unit base 300 is not particularly limited and may be an arbitrary shape.
Further, the unit chip 301 can be arranged on a part of all the side surfaces of the unit base 300 without arranging the unit chip 301 on all the side surfaces of the unit base 300.
The number of unit chips 301 held on one surface of the unit base 300 is not limited to one, and a plurality of unit chips 301 are held on one surface of the unit base 300 as shown in FIG. It may be.
Of course, the unit chip 301 may not be the same as that described in the first to third embodiments, and it is needless to say that an analysis chip having another configuration may be used as the unit chip.
[0192]
[9] Ninth embodiment
FIG. 29 is a schematic diagram for explaining a ninth embodiment of the present invention.
As shown in FIG. 29, the analysis chip unit 1H according to the ninth embodiment of the present invention has a configuration similar to that of the analysis chip according to the first to third embodiments on a flat unit base 400. A plurality of unit chips 401 are provided. The structure of the unit chip 401 described in the first, third, and seventh embodiments has been described in detail in the first, third, and seventh embodiments, and thus the description thereof is omitted.
[0193]
Further, the inlet 21 of each unit chip 401 and other unit chips corresponding to the inlet 21 except for the inlet 21 of the unit chip 401 at the most upstream and the outlet 22 of the unit chip 401 at the most downstream. The discharge port 22 of 401 is connected with the connection flow path 402 provided in the unit base 401 as shown in FIG. In this example, the discharge port 22 of one unit chip 401 and the injection ports 21 of a plurality of unit chips 401 are connected by a connection channel 402 (see reference numeral 400a in FIG. 29), or a plurality of unit chips 401. The discharge port 22 and the injection port 21 of one unit chip 401 are connected by a connection flow path 402 (see reference numeral 400b in FIG. 29), and further, the discharge ports 22 of a plurality of unit chips 401 and a plurality of units. The inlet 21 of the chip 401 is connected by a connecting channel 402 (see reference numeral 400c in FIG. 29).
[0194]
Since the analysis chip unit 1H according to the ninth embodiment of the present invention is configured as described above, the liquid sample Fs is injected from the most upstream injection port 21, and the liquid sample Fs is connected to each unit chip 401. Analysis is performed while flowing in the flow path 402. The liquid specimen Fs that has flowed through the unit chip 401 and the connection channel 402 is discharged from the discharge port 22 of the unit chip 401 that is the most downstream.
[0195]
As described above, when the analysis chip unit 1H in which the unit chips 401 are combined is used, the liquid sample Fs can be analyzed efficiently.
For example, if different types of specific substances 61 are fixed to each unit chip 401, different types of predetermined substances can be detected with only one analysis, and the liquid sample Fs can be analyzed efficiently. it can.
[0196]
Further, if the same kind of specific substance 61 is fixed to each unit chip 401, the reaction part in each unit chip 401 can be observed and measured, and a large amount of data can be obtained by performing a single analysis. Therefore, the reliability of the analysis result can be improved.
[0197]
Further, by integrating the unit chips 401 into the analysis chip unit 1H, it is possible to prevent a dead volume that occurs when using separate analysis chips and is necessary when using a plurality of analysis chips. This eliminates the need for piping, etc., and reduces the cost required for equipment preparation and inspection. Also, there is a possibility of leakage, changes in temperature and humidity outside the chip, blockage of piping, tubes, connectors, etc., tubes and connectors. It is possible to reduce things that obstruct accurate and efficient analysis, such as adsorption with materials.
[0198]
The arrangement and combination of the unit chips 401 held on the unit base 400 are not particularly limited, and can be arbitrarily set according to the analysis purpose.
Further, the unit chip 401 may not be the same as that described in the first, third to seventh embodiments, and it is needless to say that an analysis chip having another configuration may be used as the unit chip 401. Absent.
[0199]
[10] Tenth embodiment
FIG. 30 is an explanatory diagram for explaining an analyzer as a tenth embodiment of the present invention. As shown in FIG. 30, the analyzer according to the tenth embodiment of the present invention includes the analysis chips 1, 1 </ b> B, 1 </ b> C, 1 </ b> D, 1 </ b> E, 1 </ b> F (described below) described in the first, third to seventh embodiments. 1 is used as an analysis chip), an analysis unit 501 for analyzing the liquid sample Fs flowing through the analysis chip 1, and an upstream of the analysis chip 1, and the analysis chip 1 includes the liquid sample Fs. Prior to the introduction of a separation device (separation unit) 502 that separates the liquid specimen Fs by physical and / or chemical action, and a post-analysis device (post-analysis unit) that analyzes the liquid specimen Fs discharged from the analysis chip 503). Note that the analysis chips described in the first, third, and seventh embodiments have been described in detail in the first, third, and seventh embodiments, and thus description thereof is omitted.
[0200]
The type of the analysis unit 501 is arbitrary, but the analysis unit 501 normally performs analysis by any one of analysis methods of surface plasmon resonance, chemiluminescence, bioluminescence, electrochemiluminescence, fluorescence, and RI (radioisotope analysis). What to do is preferred. Note that the analysis unit may perform analysis by one of the above methods, or may perform analysis by combining two or more methods.
[0201]
When the analysis unit 501 performs analysis using surface plasmon resonance, the specific device configuration of the analysis unit 501 can be configured in the same manner as in the second embodiment described above. In particular, the analysis unit 501 using surface plasmon resonance can perform analysis by irradiating light from the back surface of the analysis chip 1. That is, the reaction part 6 formed in the flow path 5 of the analysis chip 1 is irradiated with light from the substrate 4 side of the analysis chip 1, and the reflected light from the reaction part 6 is irradiated to the substrate 4 of the analysis chip 1. Observe and analyze on the side. However, in that case, since the irradiated light needs to reach the reaction part 6 of the analysis chip 1, the substrate 4 must naturally be able to transmit the incident light. Therefore, when analyzing by irradiating light from the back surface of the analysis chip 1, the substrate 4 is usually made of a material that can transmit light having the same wavelength as the incident light.
[0202]
When the analysis unit 501 performs analysis by fluorescence, generally, the lid 2 of the analysis chip is formed transparent, and excitation light is irradiated from the lid 2 side to fluoresce from the lid 2 side. Is detected. However, as in the case of analyzing by surface plasmon resonance, the analysis can be performed by irradiating excitation light from the back side of the analysis chip 1, that is, the substrate 4 side, and detecting fluorescence on the substrate 4 side. In this case, it is necessary to form the substrate 4 transparently. Further, the fluorescence is detected on the substrate 4 side by irradiating the excitation light from the lid 2 side of the analysis chip 1, or the fluorescence is detected on the lid 2 side by irradiating the excitation light from the substrate 4 side. Is also possible.
[0203]
Even in the case where the analysis unit 501 performs analysis by chemiluminescence or bioluminescence, as in the case of using surface plasmon resonance or fluorescence, it is arbitrarily passed through the transparent part (part formed transparently) of the analysis chip 1 as appropriate. Chemiluminescence can be detected from this direction. Therefore, for example, when the lid portion 2 of the analysis chip 1 is formed transparently, light can be irradiated and detected from the lid portion 2 side. When the substrate 4 is formed transparently, from the substrate 4 side. Light irradiation and detection can be performed. In chemiluminescence and bioluminescence, irradiation with excitation light is usually unnecessary.
[0204]
The case where the analysis unit 501 performs analysis by electrochemiluminescence is almost the same as the case of analysis by chemiluminescence. However, in the case of electrochemiluminescence, it should be noted that electrodes are provided on the substrate 4. is there. Therefore, when the electrode is formed of an opaque material, it is difficult to detect electrochemiluminescence from the substrate 4 side even if the substrate 4 is formed of a transparent material. However, when the electrode is formed of a transparent material (for example, ITO), or when it is formed of an opaque material but is formed into an extremely thin thin film, light can be transmitted, the substrate 4 side It is also possible to irradiate and detect light.
[0205]
Further, in the analyzer of the present embodiment, prior to introducing the liquid sample Fs into the analysis chip 1 upstream of the analysis chip 1, a separation device that separates the liquid sample Fs by physical and / or chemical action. 502 is provided.
[0206]
The type of the separation device 502 is arbitrary, but normally, liquid chromatography or HPLC (high performance liquid chromatography) that performs separation according to the adsorptivity or distribution coefficient of the sample, or a capillary that performs separation according to the electronegativity of the sample. The use of electrophoresis, microchip electrophoresis, microchannel electrophoresis, or flow injection is suitable, but of course, other devices may be attached to the analyzer as the separation device 502, or a combination of the above devices. May be used.
[0207]
A microchannel is a groove where a sample formed on a chip surface flows, and microchannel electrophoresis is used to fill a part of this groove with an equivalent of an HPLC column packing material or to add a functional group to the groove surface. Separation is possible by providing it.
Flow injection is a technique that causes various reactions while the sample is flowing. For example, complex injection and solvent extraction are performed to remove substances other than the detected species in the sample. Separation can be performed.
Of course, a device other than the above may be attached to the analyzer as the separation device 502.
[0208]
Further, the analyzer of the present embodiment includes a post-analyzer 503 that analyzes the liquid specimen Fs discharged from the analysis chip. The type of the post-analysis device 503 is not particularly limited, and an arbitrary analysis device can be used as the post-analysis device 503. Specific examples include an MS (mass spectrometer), a protein sequencer, a DNA sequencer, SEM, SPM, Examples include STM and AFM.
The post-analysis device 503 may include a pre-processing mechanism that makes the liquid specimen Fs ready for analysis. Moreover, you may use combining said apparatus.
[0209]
Since the analyzer according to the tenth embodiment of the present invention is configured as described above, at the time of analysis, the liquid sample Fs is flowed in the order of the separation device 502, the analysis chip 1, and the post-analysis device 503, and the analysis is performed. .
Since the analysis is performed using the analysis chip 1 when the analysis is performed by the analysis unit 501, the liquid specimen Fs can be efficiently and accurately performed.
[0210]
In addition, since the separation device 502 is provided, predetermined substances such as enzymes and proteins can be separated into pure substances by the separation device in advance. For this reason, it is possible to analyze a predetermined substance that has become a pure substance, and it is possible to perform a more accurate analysis.
[0211]
Furthermore, since the post-analysis device 503 is provided, a large amount of data can be obtained by a single analysis operation, and the liquid specimen Fs can be analyzed in a multifaceted manner.
[0212]
In this embodiment, the analysis chip 1 described in the first embodiment is used as the analysis chip. However, the analysis chip does not have to be the same as this, and other configurations such as third to third are possible. It goes without saying that an analysis chip having the configuration according to the seventh embodiment may be used.
[0213]
As shown in FIG. 31, the analysis chip units 1G and 1H described in the eighth or ninth embodiment may be used instead of the analysis chip used in this embodiment. In FIG. 31, the same reference numerals as those in FIG. 30 are shown.
With such a configuration, when the analysis is performed by the analysis unit 501, the analysis is performed using the analysis chip units 1G and 1H, so that the liquid specimen Fs can be efficiently and accurately performed.
[0214]
In addition, since the separation device 502 is provided, predetermined substances such as enzymes and proteins can be separated into pure substances by the separation device in advance. For this reason, it is possible to analyze a predetermined substance that has become a pure substance, and it is possible to perform a more accurate analysis.
[0215]
Furthermore, since the post-analysis device 503 is provided, a large amount of data can be obtained by a single analysis operation, and the liquid specimen Fs can be analyzed in a multifaceted manner.
[0216]
[11] Other
The first to tenth embodiments of the present invention have been described above, but the present invention is not limited to these embodiments, and various modifications can be made without departing from the spirit of the present invention. .
For example, you may implement combining the structure of 1st-10th embodiment, respectively. In particular, the above-described fourth to seventh embodiments have been described with a focus on the configuration of the analysis chip of the third embodiment, but the configurations of other embodiments such as the first embodiment and the second embodiment have been described. You may implement in combination.
Further, for example, the second embodiment may be merged with the third embodiment. That is, the lid 2 and the plate 10 according to the analysis chip 1B of the third embodiment are made of a transparent material, and the diffraction grating 42 and the metal layer 41 are formed on the surface of the substrate 4 on which the specific substance 61 is fixed. And configured as a sensor chip. Thereby, the substrate 4 is irradiated with light through the lid 2 and the plate 10, and the light intensity of the reflected light from each specific substance 61 in the reaction unit 6 is detected, similarly to the effect of the second embodiment. The concentration of the detected species in the test fluid can be measured in real time.
[0217]
  Further, in each of the above embodiments, the means for transporting the liquid specimen Fs is constituted by a liquid feed pump. However, the means for transporting the liquid specimen Fs is not limited to this and is of a pressure type other than the liquid feed pump.Can also be used. Also,An electric field is applied to the slit-shaped flow paths 9a (first and second embodiments) and 10a (third embodiment) and the collecting flow path portions 81 and 82 (first and second embodiments), 13 and 14 (third embodiment). Addition causes the flow of liquid specimen Fs (electroosmotic flow)LetAlso good.
[0218]
Further, in each of the above-described embodiments, the example in which the liquid specimen Fs is water-soluble has been described, but the liquid specimen Fs may be oil-based.
Further, a cross section with an aspect ratio (vertical dimension / horizontal dimension) of 0.005 (for example, 5 μm in length, 1 mm in width) to 100 (for example, 1000 μm in length, 10 μm in width), and 5 mm²A plurality of flow paths having the following cross-sectional areas are provided in parallel between one injection port and one discharge port, and specific or non-specific interaction with a predetermined substance in each of the flow paths described above You may make it have a substance. Further, the cross-sectional area is preferably 100 μm.²More than 5mm²Or less, more preferably 2000 μm²0.3 mm or more²It should be: Even if comprised in this way, generation | occurrence | production of the bubble in a flow path can be suppressed, and since analysis can be performed on the optimal conditions which a bubble does not generate | occur | produce, the accuracy of an analysis result is improved. Is possible.
[0219]
Moreover, you may comprise an internal flow path over several intermediate | middle plates. That is, for example, as shown in FIGS. 32A to 32D, a first plate 8 ′ having a slit-like hole 8a ′ divided by a partition wall 8b ′ between the lid portion 2 and the substrate 4; The slit-like holes 8a 'and 8b' and the slit-like hole 9a 'together form an internal flow path with the second plate 9' having the slit-like hole 9a 'divided by the partition wall 9b'. It may be configured.
[0220]
Alternatively, the partition wall (convex member) 9b ′ may be formed directly on the substrate 4 as shown in FIG. 33A by screen printing, inkjet printing, or coating. As described in the first embodiment, when the convex member is formed by using the intermediate plate 9 ′, the convex member 9b ′ is formed as a slit-like hole as shown in FIG. 33 (b), for example. It may be difficult to assemble the analysis chip because it is separated from the intermediate plate 9 'by 9a' or the like. However, if the convex member 9b 'is formed directly on the substrate 2, the convex member 9b' can be easily provided even in such a case. Furthermore, this technique may be applied to other embodiments including the first and third embodiments.
[0221]
Further, as described in FIG. 17, the inlet 21 and the outlet 22 may be formed on the side surface of the analysis chip 1 as shown in FIG. 34. Here, of the opposing side surfaces of the plate 8, a hole 21 is formed as an injection port on one side surface, and a hole 22 is formed as a discharge port on the other side surface. Thus, for example, when analysis is performed using a surface plasmon resonance sensor, it is not necessary to provide a connector for supplying or discharging the liquid specimen Fs to the light passage portion above the analysis chip 1. In addition, it is possible to obtain an advantage that an optical system such as a light source and a detector can be brought close to the analysis chip.
[0222]
Moreover, the shape and arrangement | positioning of a convex-shaped member are not limited to what was demonstrated by embodiment mentioned above, It can be set as arbitrary shapes and arrangement | positioning. Hereinafter, a specific example thereof will be described with reference to FIGS. 35A to 35F, which are schematic views of the shape of the flow path 5.
For example, as shown in FIG. 35 (a), it may be formed in an intermittent wall shape formed intermittently as the convex member 51, or a cylindrical convex shape as shown in FIG. 35 (b). A large number of the shaped members 51 may be formed. Furthermore, as shown in FIG. 35 (c), only one convex member 51 may be formed at the center of the flow path 5, and as shown in FIG. 35 (d), the convex member ( The wall-shaped member 51 may be formed so as to be in contact with the wall surface of the flow path 5. Furthermore, the size of each convex member (wall-shaped member) 51 and the width of the internal flow path 52 may be different as shown in FIG. 35 (e), and the convex member as shown in FIG. 35 (f). The (wall forming member) 51 may not be formed to have a uniform width in the flow direction of the internal flow path 52, or may be formed without regularity.
[0223]
In each of the above-described embodiments, the convex member is formed as a partition wall that functions as a support member. However, the convex member may have a function other than the support member or the partition wall. For example, the projecting member may be a microfluidic device such as a micromixer, a micropump, a heat exchanger (heater, cooler), or a microinjector manufactured by a microfabrication technology, a MEMS technology, a semiconductor manufacturing technology, or the like.
[0224]
In the above-described embodiments, the shape of the flow channel 5 of the analysis chip has been described as a substantially hexagonal sheet shape, but the flow channel 5 may be formed in other shapes. For example, it may be a shape composed only of a curve or a shape other than a hexagonal shape. Furthermore, it may be formed in a flow path having a shape other than a sheet shape such as a cylindrical shape (a flow path having a shape other than the flow path formed in the sheet-like space).
[0225]
In addition, it can be said from the said embodiment that said convex-shaped member (partition wall) comprises a bubble suppression part means if it pays attention to the function which suppresses generation | occurrence | production of the bubble 201, and also the flow path 5 If attention is paid to the function of dividing the flow path into a plurality of internal flow paths, it can be said that the flow path dividing means for dividing the flow path 5 is constituted. In addition, the convex member (partition wall) can be said to constitute a deformation suppressing means when attention is paid to the function of suppressing the deformation of the analysis chips 1, 1 </ b> A to 1 </ b> H. If attention is paid to the function of reducing the amount, it can be said that the liquid sample reducing means is constituted.
[0226]
【Example】
Hereinafter, the present invention will be specifically described with reference to examples. However, the present invention is of course not limited to these examples, and can be implemented in various forms as long as the gist thereof is not exceeded. It is.
[0227]
An uneven shape having a groove pitch of about 870 nm and a groove depth of about 40 nm is formed on the flat resin, and the thin film layer formed of gold having a thickness of about 80 nm is formed on the surface of the flat resin using the uneven shape as a diffraction grating. It was formed by vapor deposition. This flat resin was cut into 25 mm × 25 mm to form a substrate.
[0228]
Subsequently, a photoresist NanoXP SU-8 (50) (manufactured by MicroChem Corporation) was spin-coated on a 4-inch silicon wafer (manufactured by Furuuchi Chemical Co., Ltd.), and then the heated solvent was removed for 30 minutes and cooled to room temperature. UV exposure was performed through a film mask (Falcom). At this time, as shown in FIG. 46, the photo film mask has a pattern P1 having a slit-like structure as an analysis chip having a convex member and a pattern P2 having no slit-like structure as a conventional analysis chip. Are formed on the same silicon wafer 601. These patterns P1 and P2 each have a shape corresponding to the flow path portion of the analysis chip when the analysis chip is completed, and the maximum length in the flow direction is 10 mm and the length in the width direction. The maximum length is 21 mm. Further, in the pattern P1 having the slit-like structure, the slit pattern is formed so that the flow path is divided into the internal flow paths having a width of 0.5 mm.
[0229]
After the exposure, after-baking was performed for 30 minutes, followed by development for 15 minutes by a developer (Nano XP SU-8 Developer, manufactured by MicroChem Corporation), and finally washing with isopropyl alcohol and water.
Furthermore, this agent-curing agent ratio was set to 10: 1 using a silicone elastomer PDMS (polydimethylsiloxane) Sylgard 184 kit manufactured by Toray Dow Corning Co., Ltd. and then deaerated under vacuum at -630 Torr for 15 minutes. .
[0230]
As shown in FIG. 47, a U-shaped mold 602 made of PMMA having a thickness of 1 mm and a resin flat plate 603 having a thickness of 1 mm are stacked on the silicon wafer 601 to form an elastomer-filled portion. From the above, the elastomer was filled and then cured at 80 ° C. for 3 hours. After curing, the elastomer was peeled from the silicon wafer 601 and the U-shaped mold 602. After curing, portions corresponding to the portions where the patterns P1 and P2 were formed were cut out as sheet-like flow path portions, and through holes serving as inlets and outlets were formed in the resin flat plate 603. Pyrex (registered trademark) glass having a thickness of 1 mm in which through holes that serve as inlets and outlets are formed as lids in the cut portions so that the through holes in the lid and the through holes in the flow cell are aligned. Finally, by combining with the substrate and combining with the substrate, the one corresponding to the portion where the pattern P1 having the slit-like structure is formed is completed as an analysis chip having a convex member, A chip corresponding to the portion where the pattern P2 having no slit-like structure was formed was completed as a conventional analysis chip. In addition, the depth of the flow path portion was created to be 90 μm in both analysis chips.
[0231]
Using each of the analysis chip having a convex member and the conventional analysis chip prepared as described above, the SPR measurement device FLEX CHIP^TM  SPR analysis was performed using a Kinetic Analysis System (HTS Biosystems). The measurement is performed at 30 ° C., and pure water is circulated at a constant flow rate of 500 μl / min for 60 minutes in the flow channel of the analysis chip, and at that time, the resonance angle changes 512 points every about 7 seconds. I got it. Moreover, the measurement area provided 8 * 20 = 160 micro measurement area | regions (ROI) in the detection area of 10 mm x 10 mm, and acquired each value. The difference between the maximum value and the minimum value in the measurement time range (60 minutes) for each minute measurement region was defined as the drift amount, and the analysis chip having a convex member was compared with the conventional analysis chip. FIG. 48 (a) shows the drift amount (angle difference) when an analysis chip having a convex member is used, and FIG. 48 (b) shows the drift amount (angle difference) using a conventional analysis chip. . Table 1 shows the average drift amount for each minute measurement region.
[0232]
[Table 1]

[0233]
From Table 1, it can be seen that the analysis chip having the convex member has a smaller drift amount than the conventional analysis chip. As a result, an analysis chip having a convex member can perform highly accurate analysis, and can reduce the frequency of redoing the analysis work and the amount of liquid sample used in the analysis work, thereby making analysis more efficient. It was confirmed that it can be done.
[0234]
【The invention's effect】
As described above in detail, according to the analysis chip, the analysis chip unit, and the analysis apparatus of the present invention, a liquid sample can be analyzed efficiently and accurately, and any of the following effects can be achieved. Obtainable.
(1) It is possible to suppress the generation of bubbles due to the surrounding of the liquid specimen.
(2) Deformation of the analysis chip can be prevented.
(3) The amount of liquid specimen can be reduced.
[0235]
Further, according to the method for producing an analysis chip of the present invention, the surface of the intermediate plate opposite to the substrate is more specific to the liquid containing the specific substance than the wall surface of the inner hole of the intermediate plate and / or the surface of the substrate on the flow path side. An intermediate plate composed of members with low affinity and formed with a plurality of slit-shaped holes is fixed on the substrate, and then a specific substance-containing liquid is dropped onto the substrate through the slit-shaped holes of the intermediate plate, and the specific substance is placed on the substrate. Since the lid is fixed on the intermediate plate after fixing it in a spot shape, even if the specific substance-containing liquid is applied to the partition wall of the intermediate plate, the specific substance-containing liquid is not the same as the substrate of the intermediate plate. It flows (induces) to the substrate through the wall surface of the internal hole and / or the flow path side surface of the substrate having higher affinity for the specific substance-containing liquid than the opposite surface. That is, the specific substance can be stably fixed at a target position on the substrate, and the analysis chip on which the specific substance is fixed can be easily and accurately manufactured.
[0236]
[Brief description of the drawings]
1A and 1B are diagrams showing an analysis chip as a first embodiment of the present invention, in which FIG. 1A is a schematic assembly perspective view thereof, and FIG. 1B is a schematic exploded perspective view thereof;
2A and 2B are schematic views showing an analysis chip as a first embodiment of the present invention, in which FIG. 2A is a YY sectional view of FIG. 1A, and FIG. X1-X1 sectional drawing of this, (c) is X2-X2 sectional drawing of Fig.1 (a).
3A and 3B are schematic views showing the analysis chip as the first embodiment of the present invention, in which FIG. 3A is a top view of the lid portion, FIG. 3B is a top view of the intermediate plate, and FIG. Is a top view of the intermediate plate, and (d) is a top view of the substrate.
FIG. 4 is a schematic diagram for explaining a definition of a flow direction of a liquid specimen.
FIG. 5 is a schematic top view for explaining a method for producing an analysis chip as a first embodiment of the present invention.
6A is a plan view schematically showing a flow path formed in a conventional sheet-shaped space, and FIG. 6B is a slit-shaped flow path of the analysis chip according to the first embodiment of the present invention. It is a top view which shows typically.
7A and 7B are schematic views showing a first modification of the first embodiment of the present invention, in which FIG. 7A is a bottom view of a lid portion, FIG. 7B is a top view of an intermediate plate, and FIG. FIG.
8A and 8B are diagrams showing a second modification of the first embodiment of the present invention, in which FIG. 8A is a bottom view of a lid portion, and FIG. 8B is a top view of a substrate.
FIGS. 9A and 9B are schematic views showing a third modification of the first embodiment of the present invention, where FIG. 9A is a top view of a lid portion, FIG. 9B is a top view of an intermediate plate, and FIG. 9C is a substrate; FIG.
FIG. 10 is a schematic perspective view showing an overall configuration of an SPR sensor according to a second embodiment of the present invention.
FIG. 11 is a schematic exploded perspective view showing a configuration of an analysis chip as a second embodiment of the present invention.
12A and 12B are diagrams showing an analysis chip as a third embodiment of the present invention, in which FIG. 12A is a schematic assembly perspective view thereof, and FIG. 12B is a schematic exploded perspective view thereof.
FIG. 13 is a schematic bottom view showing an intermediate plate provided in an analysis chip as a third embodiment of the present invention.
14A and 14B are schematic views showing an analysis chip as a third embodiment of the present invention, where FIG. 14A is a cross-sectional view taken along the line Y1-Y1 in FIG. 12A, and FIG. It is X3-X3 sectional drawing of.
FIGS. 15A and 15B are schematic views showing a first modified example of the analysis chip as the third embodiment of the present invention, in which FIG. 15A is a bottom view of a lid portion and FIG. 15B is a top view of a substrate; .
FIGS. 16A and 16B are schematic views showing a second modified example of the analysis chip as the third embodiment of the present invention, wherein FIG. 16A is a bottom view of the lid portion and FIG. 16B is a top view of the substrate; .
FIGS. 17A and 17B are schematic cross-sectional views showing a third modification of the analysis chip as the third embodiment of the present invention. FIGS.
FIG. 18 is a schematic bottom view showing a modification of the intermediate plate according to the third embodiment of the present invention.
FIGS. 19A and 19B are schematic views showing an analysis chip according to a fourth embodiment of the present invention, in which FIG. 19A is a bottom view of an intermediate plate, FIG. 19B is a bottom view of a lid portion, and FIG. It is a top view.
20A and 20B are schematic views showing an intermediate plate according to a fifth embodiment of the present invention, in which FIG. 20A is a bottom view of the intermediate plate, and FIG. 20B is an enlarged view of the XXb portion of FIG. (C) is a figure which expands and shows the principal part of the conventional plate.
FIG. 21 is a schematic view showing an analysis chip according to a sixth embodiment of the present invention, in which (a) shows a part of the flow path of the analysis chip in which the partition wall is formed in the width direction of the flow path; Sectional drawing cut | disconnected by the surface orthogonal to (b), (b) is a figure which expands and shows the XXIb part of (a).
FIG. 22 is a schematic bottom view showing an intermediate plate according to a seventh embodiment of the present invention.
FIGS. 23A and 23B are schematic views for explaining a seventh embodiment of the present invention. FIG.
FIG. 24 is a schematic bottom view showing an intermediate plate according to a first modification of the seventh embodiment of the present invention.
FIG. 25 is a schematic bottom view showing an intermediate plate according to a second modification of the seventh embodiment of the present invention.
FIGS. 26A, 26B and 26C are schematic bottom views showing intermediate plates according to third to fifth modifications of the seventh embodiment of the present invention, respectively.
FIG. 27 is a schematic perspective view of an analysis chip unit according to an eighth embodiment of the present invention.
FIG. 28 is a schematic perspective view of an analysis chip unit according to a modification of the eighth embodiment of the present invention.
FIG. 29 is a schematic plan view for explaining an analysis chip unit according to a ninth embodiment of the present invention.
FIG. 30 is a schematic diagram for explaining an analyzer according to a tenth embodiment of the present invention.
FIG. 31 is a schematic diagram for explaining an analyzer according to a modification of the tenth embodiment of the present invention.
32A and 32B are schematic views showing an embodiment of the present invention, in which FIG. 32A is a top view showing a lid, FIG. 32B is a top view showing a first plate, and FIG. 32C is a second plate; (D) is a top view showing a substrate.
33A and 33B are schematic views showing an analysis chip according to an embodiment of the present invention, in which FIG. 33A is an exploded perspective view thereof, and FIG. 33B is a cross-sectional view cut along a plane perpendicular to the flow direction of the flow path. It is sectional drawing.
FIG. 34 is a schematic assembly perspective view of an analysis chip showing an embodiment of the present invention.
FIGS. 35A to 35F are schematic views for explaining an example of the flow path of the analysis chip of the present invention.
36 (a) and 36 (b) are schematic views for explaining a conventional analysis chip.
FIG. 37 is a schematic diagram for explaining a conventional analysis chip.
FIGS. 38A and 38B are schematic views for explaining a conventional analysis chip.
FIGS. 39A and 39B are schematic diagrams for explaining a conventional analysis chip.
FIG. 40 is a diagram illustrating a state in which a liquid specimen is advanced.
FIG. 41 is a diagram for explaining the configuration of a flow path for preventing the advance of a liquid specimen.
FIG. 42 is a diagram for explaining the configuration of a flow path for preventing the advance of a liquid specimen.
FIG. 43 is a cross-sectional view of the flow path for explaining the configuration of the flow path for preventing the advance of the liquid specimen.
FIG. 44 is a diagram for explaining a configuration of a flow path for preventing a liquid specimen from being advanced.
45 is a view showing a modification of the first embodiment of the present invention, where (a) is a top view of the lid, (b) is a top view of the plate, and (c) is a top view of the substrate. FIG. is there.
FIG. 46 is a diagram for explaining an example of the present invention.
FIG. 47 is a diagram for explaining an example of the present invention.
48A is a graph showing the results of an example of the present invention, and FIG. 48B is a graph showing the results of using a conventional analysis chip.
[Explanation of symbols]
1,1A, 1B, 1C, 1C, 1E, 1F Analysis chip
1G, 1H analysis chip unit
2 lid
2a Slit groove
2b partition wall
4 Substrate
4a Slit channel
4b partition wall
5,5 'flow path
5a Long side of cross section of flow path 5
5b Short side of cross section of flow path 5
5x hydrophobic part (second affinity part)
5y hydrophilic part (hydrophilic part, first affinity part)
6 reaction part
8,8 'plate (intermediate plate)
9,9 'plate (intermediate plate)
8a ', 9a, 9a' Slit hole (slit channel)
8b ', 9b, 9b' partition wall
10 plates (intermediate plates)
10a Slit groove (slit channel)
10b partition wall
10c Constriction section
10t tetrafluoroethylene layer
11,12 holes
13, 14 recess
21 holes (inlet)
21 ', 22' Groove (aggregate flow path)
26 and 27 pipes
22 holes (discharge port)
23, 24 Groove
41 Metal layer
42 Diffraction grating
43, 44, 46, 47 Groove (concave)
51 Convex member
52, 52a-e Internal flow path
61 Specific substances
81 holes (collecting flow path)
81x Upstream end of hole 81
82 holes (collection flow path)
82x downstream end of hole 82
83 Slit hole
91 Upstream end of slit hole (slit channel)
92 Downstream end of slit hole (slit channel)
91 ', 92' hole
100 light sources
101 detector
200 gas
201 bubbles
202 Particulate matter
204 reaction area
204a In the vicinity of the bubble 201
205 Measurement area
300,400 unit base
302 Rotation support shaft
301,401 unit chip
402 Connection flow path
501 Analysis Department
502 Separator
503 Post-analyzer
601 Silicon wafer
602 U-shaped mold
603 resin flat plate
Fs liquid specimen
S, St1-St4 solid-gas-liquid three-phase boundary line
W Channel width
H Channel height
P1 Pattern with slit-like structure
P2 Pattern without slit structure

Claims (24)

Analyzing the liquid specimen based on the interaction between a predetermined substance and a specific substance fixed facing the flow path by circulating the liquid specimen in a flow path having a closed cross-sectional structure by a pressure-type transport means In the analytical chip used to do
An analysis chip characterized in that the flow path has a strut member interposed between opposing surfaces of the flow path . The analysis chip according to claim 1, wherein the flow path is formed in a sheet-like space. One inlet for injecting the liquid specimen, provided at the upstream end of the flow path;
The analysis chip according to claim 1, wherein the analysis chip is provided at a downstream end portion of the flow path and is provided with a single discharge port for discharging the liquid specimen. A substrate,
A lid member;
Comprising at least one intermediate plate interposed between the substrate and the lid member, and forming a sheet-like space having the flow path in cooperation with at least one of the substrate and the lid member,
Strut member, characterized in that it is interposed between the mutually facing surfaces of at least one of the intermediate plate and the substrate and the cover member in the flow path in the sheet-like space, claim 1 The analysis chip according to any one of -3 . A substrate,
A lid member disposed opposite to the substrate and forming a sheet-like space having the flow path in cooperation with the substrate;
Strut member, characterized in that it is interposed between the surfaces facing each other between the substrate and the lid member in the flow path in the sheet-like space, one of claims 1 to 3 1 The analysis chip according to item . In addition to the floor and ceiling surfaces constituting the flow path, the left side surface, the right side surface, the upstream end surface, and the downstream end surface form the sheet-like space,
The support member, between the left side and right side of the, and, characterized in that it is interposed at least either between the upstream side end surface and the downstream end surface of the above claims 4 Alternatively, the analysis chip according to claim 5 . Strut member, characterized in that it is in direct contact with the opposite faces, the analysis chip according to any one of claims 1 to 6. A portion of the strut member is in direct contact with one of the opposing surfaces;
The other end of the strut member, when allowed to flow fluid flow path, the other of the opposed surfaces, characterized in that it is in contact through the fluid, one of the claims 1 to 6 2. The analysis chip according to item 1. 9. The analysis chip according to claim 8 , wherein an adhesion reducing layer is formed on a surface of the support member. The analysis chip according to any one of claims 1 to 9 , wherein the specific substance is fixed in the flow path. The flow path is provided with a first affinity part and a second affinity part having a lower affinity for the liquid specimen than the first affinity part, respectively. The analysis chip according to any one of 10 . The specific substance is immobilized on the surface of the flow path,
12. The analysis according to claim 11 , wherein the first affinity part and the second affinity part are provided upstream of the portion where the specific substance is immobilized in the flow direction of the flow path. For chips. Affinity of affinity portion and the second first, respectively, characterized in that it is formed in a band shape extending in a direction intersecting the flow direction of the flow path, to claim 11 or claim 12 The analysis chip described. The analysis chip according to any one of claims 11 to 13 , wherein a plurality of the first affinity parts and the second affinity parts are alternately arranged. Affinity of the first are hydrophilic portion, wherein the affinity of the second is a hydrophobic unit, the analysis chip according to any one of claims 11 to 14. The analysis chip according to any one of claims 11 to 15 , wherein the first affinity part is a rough surface part, and the second affinity part is a smooth surface part. A surface on which the specific substance is fixed to the flow path;
On the surface,
A diffraction grating that generates an evanescent wave by light irradiation;
The analysis chip according to any one of claims 1 to 16, wherein a metal layer capable of inducing surface plasmon waves is provided. The analytical chip according to any one of claims 1 to 17 , wherein the analytical chip is made of a material having a Young's modulus of 60 GPa or more and 1000 GPa or less. An analysis chip comprising a unit base having a plurality of surfaces, the analysis chip according to any one of claims 1 to 18 being provided as a unit chip on the surface of the unit base. unit. With unit base,
A plurality of analysis chips according to any one of claims 1 to 18 are provided as unit chips on the unit base,
An analysis chip unit, characterized in that a connection flow path for connecting corresponding unit chips among the plurality of unit chips is provided. The analysis chip according to any one of claims 1 to 18 , or the analysis chip unit according to claim 19 or 20 ,
An analysis apparatus comprising: an analysis unit that analyzes a liquid specimen. The analysis unit performs an analysis by an analysis method using at least one method selected from the group consisting of surface plasmon resonance, chemiluminescence, bioluminescence, electrochemiluminescence, fluorescence, and radioisotope analysis. The analyzer according to claim 21 , characterized in that The separation part which isolate | separates this liquid sample by a physical and / or chemical action prior to introduce | transducing this liquid sample into this analysis chip or this analysis chip unit, or 21 characterized by the above-mentioned. 22. The analyzer according to 22 . The analyzer according to any one of claims 21 to 23 , further comprising a post-analysis unit that analyzes the liquid sample discharged from the analysis chip or the analysis chip unit.

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