WO2016043196A1

WO2016043196A1 - Sample analysis chip

Info

Publication number: WO2016043196A1
Application number: PCT/JP2015/076190
Authority: WO
Inventors: 小澤　知之
Original assignee: 凸版印刷株式会社
Priority date: 2014-09-16
Filing date: 2015-09-15
Publication date: 2016-03-24
Also published as: JP2016061610A; JP6323274B2

Abstract

　This sample analysis chip is provided with: a substrate having a plate-shaped section sandwiched by a first surface and a second surface; a first reaction chamber provided to the plate-shaped section to store a sample liquid and cause a first reaction to occur in the entire sample liquid; a measurement chamber communicated with the first reaction chamber and formed in the plate-shaped section, the measurement chamber having a plurality of branching paths for measuring out a first reaction liquid obtained by the first reaction occurring in the sample liquid in the first reaction chamber; and a second reaction chamber group having a plurality of second reaction chambers communicated with the plurality of branching paths and formed in the plate-shaped section in order to individually cause a second reaction in the first reaction liquid moved through the plurality of branching paths; any one chamber among the first reaction chamber, the measurement chamber, and the second reaction chamber group being provided to the first surface, and the chambers not provided to the first surface among the first reaction chamber, the measurement chamber, and the second reaction chamber group being provided to the second surface.

Description

Sample analysis chip

The present invention relates to a sample analysis chip. For example, the present invention relates to a sample analysis chip that is used for detection and analysis of biochemical reactions and can perform a plurality of processes.
This application claims priority based on Japanese Patent Application No. 2014-188124 filed in Japan on September 16, 2014, the contents of which are incorporated herein by reference.

Conventionally, in the field of biochemical reactions such as DNA reactions and protein reactions, techniques called μ-TAS (Total Analysis System) and Lab-on-Chip are known as reaction apparatuses for processing a small amount of sample solution. . This is a reaction apparatus provided with a plurality of reaction chambers (hereinafter referred to as wells) and flow paths in one chip or cartridge, and can analyze a plurality of specimens or perform a plurality of reactions. These techniques have been considered to have various merits by reducing the amount of chemicals handled by reducing the size of the chip and cartridge.

The benefits include, for example, the fact that the amount of strong acids and strong alkaline chemicals that have been used in the past has been reduced to a much lower level, and the impact on the human body and the environment will be significantly reduced. For example, the amount of reagents consumed can be reduced so that the cost of analysis and reaction can be reduced.

In order to perform biochemical reactions most efficiently using chips and cartridges, reagent reagents that react with different chemicals, specimens, and enzymes are placed in multiple wells and react with these chemicals, specimens, and enzymes. Must flow from one or several main conduits into the well to produce different reactions.
By using the reaction apparatus as described above, a plurality of types of specimens can be simultaneously processed with the same reagent, and conversely, a single type of specimen can be simultaneously subjected to a plurality of processes. For this reason, it is possible to greatly reduce the time and labor required in the past.

However, the samples poured at this time are often expensive and valuable samples, and some biochemical reaction processes are often performed first. For this reason, there are many usage methods in which a reaction is performed in a well using a sample that has been processed once in advance.

In other words, when using this type of reaction device, the technology that processes the entire sample first, the technology that sends the same amount of sample to multiple reaction fields, and the contents of each well after feeding are not mixed. Three technologies to make are important. The following prior arts can be cited as prior arts for chips that perform liquid feeding to such wells.

Patent Document 1 describes a technique for deforming and sealing a flow path in order to make a well independent in a chip that sends liquid from a liquid reservoir to a well using centrifugal force.
Patent Document 2 describes a technique of reducing variation in the amount of liquid fed to each well by interweaving rotation and revolution when a centrifugal force is applied.
Patent Document 3 describes an analysis medium in which a plurality of wells each having a liquid storage part and a channel extending in the centrifugal direction are connected. However, the liquid distribution property of the liquid is not closely watched. The fluid is controlled by pressing the air and liquid clogged in the well.
According to Patent Document 4, a heat-meltable substance that has been preliminarily filled in a microchannel is moved by air pressure from the inlet while controlling the viscosity with a heater, and fluid control in the microchannel is performed. A method is described.

Japanese Special Table 2004-502164 Japanese Patent No. 3699721 Japanese Unexamined Patent Publication No. 2008-83017 US Pat. No. 7,195,036

However, the prior art as described above has the following problems.
The technique described in Patent Document 1 requires a mechanism for crushing the flow path, and is difficult to automate. In addition, when the centrifugal liquid feeding is performed from the central liquid reservoir to the surrounding wells as in the conventional centrifugal liquid feeding chip, the liquid feeding amount to each well varies.
The technique described in Patent Document 2 requires a complicated mechanism and space for the tip to rotate and revolve, and when a plurality of treatments are to be added to the sample liquid, as shown in FIG. If a plurality of processes are performed after the distribution is performed, variations in each reaction well overlap and stable processes cannot be performed.
In the technique described in Patent Document 3, the liquid in the flow path between the liquid storage part and the liquid storage part is not supplied, and the amount of liquid supplied to each well varies greatly. It will occur.
In the technique described in Patent Document 4, an air pressure control mechanism is necessary, and in a solution processing chip that performs many reactions at once, the shape and mechanism for controlling the movement of the hot-melt material are complicated. End up.

In general, when an enzyme reaction is performed, if the enzyme and the substrate come into contact before reaching the reaction start temperature, an undesirable side reaction or reaction inhibition may occur, resulting in a problem that the reaction efficiency is lowered. In the enzyme reaction on the chip, for example, when each reaction chamber is filled with the enzyme and the substrate is fed as a sample solution, the enzyme and the substrate come into contact with each other when the liquid feeding is completed, and side reactions and reaction inhibition occur. May happen.
Furthermore, when performing a biochemical reaction on a chip, the reaction is often performed by heating the inside of the reaction chamber after distributing the sample solution. Therefore, in order to prevent the sample solution from evaporating, it is sometimes necessary to inject an evaporation preventing material such as mineral oil into the chip or to physically seal the chip, which increases the number of work steps. There is.

The present invention has been made in view of the problems as described above, and can easily send and measure a sample liquid to be analyzed, and can perform a plurality of types of biochemical reactions even in a compact configuration. An object of the present invention is to provide a sample analysis chip that can be used.

In order to solve the above problems, a sample analysis chip according to one aspect of the present invention stores a base material having a plate-like portion sandwiched between a first surface and a second surface, a sample liquid, and the sample A first reaction chamber provided in the plate-like portion for causing a first reaction to the whole liquid, and communicates with the first reaction chamber, and the first reaction is performed on the sample liquid in the first reaction chamber. A plurality of branch passages for measuring the first reaction liquid obtained by the occurrence of the above, a measuring chamber formed in the plate-like portion, communicated with the plurality of branch passages, and through the plurality of branch passages A second reaction chamber group having a plurality of second reaction chambers formed in the plate-like portion in order to cause a second reaction individually to the moved first reaction liquid, and One of the reaction chamber, the measuring chamber, and the second reaction chamber group Two chambers are provided on the first surface, and among the first reaction chamber, the weighing chamber, and the second reaction chamber group, a chamber that is not provided on the first surface is provided on the second surface. Yes.

Of the first reaction chamber, the weighing chamber, and the second reaction chamber group, at least a part of the chamber provided on the first surface and the chamber provided on the second surface are plate-shaped. You may arrange | position so that it may overlap in the plate | board thickness direction of a part.

The measuring chamber may be provided on the first surface, and the first reaction chamber may be provided on the second surface.

The plurality of second reaction chambers may be provided on the second surface.

Each flow path of the plurality of branch paths provided in the measurement chamber is paired with each chamber of the plurality of second reaction chambers by a plurality of vertical holes extending in the plate thickness direction of the plate-like portion. You may be connected to one.

The first reaction chamber may have a groove portion meandering in the circumferential direction in an annular region and having an elongated shape.

The first reaction solution communicated with the first reaction chamber and the measurement chamber, and stored before the first reaction solution sent from the first reaction chamber is sent to the measurement chamber, the components of the first reaction solution A sample mixing chamber for promoting mixing may be provided.

A sample inlet having a central region surrounded by the first reaction chamber, the metering chamber, and the second reaction chamber group, and for feeding the sample liquid to the first reaction chamber in the central region And a surplus liquid storage chamber for storing surplus liquid overflowing from any of the first reaction chamber, the measurement chamber, and the second reaction chamber group.

A convex portion having a frustoconical outer shape whose diameter decreases along a protruding direction protruding from the plate-shaped portion in the central region, and in a central direction of the convex portion in a direction opposite to the protruding direction of the convex portion; The sample injection port having a tapered hole portion having a reduced diameter and capable of supporting a dispensing device is provided, and the surplus is formed by a recess formed in the convex portion in a direction opposite to the protruding direction of the convex portion. A liquid storage chamber may be formed.

The surplus liquid storage chamber may be provided with an air passage that opens to the outside.

A plurality of lid members attached to the surface of the plate-like portion may be provided so as to cover each of the first reaction chamber, the measurement chamber, and the second reaction chamber group.

The plurality of lid members may include a metal lid member provided with a metal sheet-like member.

The plurality of lid members may include a composite lid member provided with a composite material of a metal and a resin film.

A light-absorbing material may be included between the metal sheet and the resin film, and the lid member in which the metal sheet and the resin film are bonded via the light-absorbing material may be included.

The plate-like portion may be made of a resin material.

According to the sample analysis chip according to one aspect of the present invention, the first reaction chamber, the measurement chamber, and the second reaction chamber (first) are separated into the first surface and the second surface of the plate-like portion (separately). 2 reaction chamber groups), it is easy to send and measure the sample liquid to be analyzed, and it is possible to perform a plurality of types of biochemical reactions even with a compact configuration.

It is a typical front view showing the composition of the sample analysis chip concerning a 1st embodiment of the present invention. It is the top view which looked at the sample analysis chip | tip in FIG. 1 from the A direction. It is the top view which looked at the sample analysis chip | tip in FIG. 1 from the B direction. It is a typical back view of the base material used for the sample analysis chip concerning the 1st embodiment of the present invention. It is a typical top view of a substrate used for a sample analysis chip concerning a 1st embodiment of the present invention. It is a typical sectional view near the injection mouth of the sample analysis chip concerning a 1st embodiment of the present invention. It is typical sectional drawing in the connection part of the 1st reaction chamber and measurement chamber of the sample analysis chip concerning the 1st embodiment of the present invention. It is the elements on larger scale of the back view of the base material used for the sample analysis chip concerning the 1st embodiment of the present invention. FIG. 8 is a cross-sectional view taken along the line CC shown in FIG. It is typical sectional drawing near the vent hole of the sample analysis chip concerning the 1st embodiment of the present invention. It is the elements on larger scale of the top view of the base material used for the sample analysis chip concerning the 1st embodiment of the present invention. It is typical sectional drawing in the connection part of the measurement chamber of a sample analysis chip concerning the 1st Embodiment of this invention, and a 2nd reaction chamber. It is a typical top view which shows the structure of the sample analysis chip concerning the 2nd Embodiment of this invention. It is a typical front view which shows the structure of the sample analysis chip concerning the 2nd Embodiment of this invention. It is the top view which looked at the sample analysis chip in Drawing 12B from the D direction. It is a typical back view of the base material used for the sample analysis chip concerning a 2nd embodiment of the present invention. It is a typical top view of a substrate used for a sample analysis chip concerning a 2nd embodiment of the present invention. It is a typical sectional view in a vent of a sample analysis chip concerning a 2nd embodiment of the present invention. It is sectional drawing in the EE line shown to FIG. 12A. It is a typical front view which shows the structure of the sample analysis chip concerning the 3rd Embodiment of this invention. It is the top view which looked at the sample analysis chip | tip in FIG. 18 from the F direction. It is a typical exploded perspective view of the sample analysis chip concerning a 3rd embodiment of the present invention. It is a typical top view of a substrate used for a sample analysis chip concerning a 3rd embodiment of the present invention. It is a typical back view of the base material used for the sample analysis chip concerning the 3rd embodiment of the present invention. It is sectional drawing in the GG line shown to FIG. 21A.

Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings. In all the drawings, even if the embodiments are different, the same or corresponding members are denoted by the same reference numerals, and common description is omitted.

[First Embodiment]
A sample analysis chip according to the first embodiment of the present invention will be described.
FIG. 1 is a schematic front view showing the configuration of the sample analysis chip according to the first embodiment of the present invention. FIG. 2A is a plan view of the sample analysis chip in FIG. 1 viewed from the A direction. FIG. 2B is a plan view of the sample analysis chip in FIG. 1 viewed from the B direction. FIG. 3 is a schematic back view of a base material used for the sample analysis chip according to the first embodiment of the present invention. FIG. 4 is a schematic plan view of a substrate used for the sample analysis chip according to the first embodiment of the present invention. FIG. 5 is a schematic cross-sectional view in the vicinity of the inlet of the sample analysis chip according to the first embodiment of the present invention. FIG. 6 is a schematic cross-sectional view of the connection portion between the first reaction chamber and the measurement chamber of the sample analysis chip according to the first embodiment of the present invention. FIG. 7 is a partially enlarged view of a back view of the base material used in the sample analysis chip according to the first embodiment of the present invention. 8 is a cross-sectional view taken along the line CC shown in FIG. FIG. 9 is a schematic cross-sectional view of the vicinity of the vent hole of the sample analysis chip according to the first embodiment of the present invention. FIG. 10 is a partially enlarged view of a plan view of a base material used in the sample analysis chip according to the first embodiment of the present invention. FIG. 11 is a schematic cross-sectional view of a connection portion between the measurement chamber and the second reaction chamber of the sample analysis chip according to the first embodiment of the present invention.

As shown in FIGS. 1 to 3, the sample analysis chip 1 according to the present embodiment injects a sample liquid (not shown) to be analyzed in the biochemical analysis, and the first reaction and internal reaction are performed on the sample liquid. It is a substantially disk-shaped device used for performing the second reaction. Examples of the sample solution, the first reaction, and the second reaction will be described later.
In the analysis using the sample analysis chip 1, it is possible to use pressure feeding and movement using centrifugal force with respect to movement of the sample liquid. For this reason, the sample analysis chip 1 is arranged in an analysis device (not shown) such that the center of the outer shape coincides with the rotation center of the analysis device.
As shown in FIG. 1, the sample analysis chip 1 includes a chip body 2 (base material, plate-like portion), an upper lid 3 (lid material), and a lower lid 4 (lid material).

The chip body 2 is composed of a plate-like member (a plate-like member having the first surface 2f and the second surface 2g) sandwiched between the first surface 2f and the second surface 2g. The outer shape of the chip body 2 in plan view is circular.
On the surface and inside of the chip body 2, in order to introduce or move the sample liquid, a flow path structure is formed by grooves, recesses, holes and the like communicating with each other.

The material of the chip body 2 is not particularly limited as long as it can form a flow channel structure described later and does not affect the sample such as a sample liquid.
As a material of the chip body 2, for example, a resin material or a metal material can be used.
As a manufacturing method of the chip body 2, in the case of a resin material, various resin molding methods such as injection molding and vacuum molding, mechanical cutting, and the like can be used. In order to manufacture at low cost, it is preferable to use a resin molded product using a mold. Further, if the cross-sectional dimension of the flow channel structure is about a sub-millimeter order, extrusion molding or the like can be used.
In the case of a metal material, it can be formed by grinding or etching using a thick base material, or pressing or drawing a thin metal sheet (sheet-like member).

Examples of resin materials that can be used for the chip body 2 include, for example, synthetic resins such as polypropylene resin, polycarbonate resin, acrylic resin, silicon resin, and fluororesin in consideration of ease of manufacture and mass productivity. Can do.

In particular, by using a resin material containing any of a polypropylene resin, a polycarbonate resin, and an acrylic resin, good visible light permeability can be ensured. For this reason, it is possible to use an optical detection method for the detection of the reaction in the first reaction or the second reaction.
In the present specification, “transparent” and “light transmittance” mean that the average transmittance in the wavelength region of the detection light is 70% or more.
The wavelength range of the detection light is not limited to the visible light region (wavelength 350 to 780 nm). If a material having optical transparency in the visible light region is used as the chip body 2, the sample state in the chip can be easily viewed. When a detector having sensitivity to detection light other than the visible light region is used as the detector, it is sufficient that the transmittance of only the wavelength region of the detection light is good.

As the polypropylene resin, for example, homopolypropylene or a random copolymer of polypropylene and polyethylene can be used.
Moreover, as an acrylic resin, the copolymer of monomers, such as polymethyl methacrylate or methyl methacrylate, and other methacrylic acid ester, acrylic acid ester, styrene, can be used, for example.
Moreover, when using these resin materials, the heat resistance and intensity | strength of a chip | tip can also be ensured.

Examples of the metal material that can be used for the chip body 2 include metal materials such as aluminum, copper, silver, nickel, and brass.
When a metal material is used as the material of the chip body 2, the thermal conductivity is good and the sealing performance can be further improved.

In the present embodiment, as an example, the chip body 2 uses a base material made of an ultraviolet curable resin using an additive manufacturing method.

The upper lid 3 is a disc-shaped lid member that is attached to the first surface 2f of the chip body 2 and seals the flow path structure formed on the first surface 2f.
However, as shown in FIG. 2A, an injection port 3a (sample injection port) composed of a circular hole for injecting a sample liquid into the chip body 2 and a flow channel structure in the chip body 2 are provided at the center of the upper lid 3. A vent hole 3b made of a circular hole for discharging a fluid such as air introduced into is provided so as to penetrate in the thickness direction.

The lower lid 4 is a disc-shaped lid member that is attached to the second surface 2g of the chip body 2 and seals the flow path structure formed on the second surface 2g. The lower lid 4 is made of a disk-shaped member having a smaller diameter than the outer shape of the chip body 2.

The material of the upper lid 3 and the lower lid 4 may be any material that can be attached in close contact with the surface of the chip body 2 and can seal a fluid such as a sample solution introduced into the channel structure of the chip body 2. Any suitable material can be used.
For example, as the upper lid 3 and the lower lid 4, for example, a metal sheet (metal sheet, metal lid member) such as aluminum or stainless steel can be used. As a method of attaching the upper lid 3 and the lower lid 4 to the chip body 2, for example, adhesion, welding, or the like can be used.
Thus, when a metal sheet is used as the upper lid 3 and the lower lid 4, the thermal conductivity is better than when the upper lid 3 and the lower lid 4 are made of a resin material or the like. Therefore, in order to control the reaction of the sample liquid in the flow channel structure, when the sample analysis chip 1 is heated or cooled, it can be quickly and accurately heated and cooled.

Further, as the upper lid 3 and the lower lid 4, a composite lid material made of a composite material of a metal and a resin film can be used. For example, when the chip body 2 is made of a resin, the upper lid 3 and the lower lid 4 are constituted by a laminate of a metal sheet and a resin film, a metal layer is deposited on the surface of the resin film, or a resin layer is applied to the metal sheet. It is possible to apply.
In this case, by selecting a resin material having a melting point close to that of the material of the chip body 2 as the material of the resin film, the resin film is brought into close contact with the chip body 2 and heat is applied from the metal side (surface) having good thermal conductivity. In addition, the upper lid 3 and the lower lid 4 can be easily attached to the chip body 2 by heat welding or the like.
Also, a laser that performs laser irradiation after applying and adhering a material that absorbs an arbitrary light wavelength (light absorbing material) such as carbon powder or an infrared absorber between a metal sheet and a resin film It is also possible to fix the metal sheet and the resin film by welding (via a light absorbing material).
In this case, by appropriately selecting the wavelength of the light-absorbing material, it is possible to absorb light having a wavelength other than that to be observed when analyzing the biochemical reaction. In this way, measurement sensitivity during analysis can be improved.

The materials of the upper lid 3 and the lower lid 4 may be the same, or different materials may be used as necessary.
For example, in this embodiment, as described later, a reaction that requires heating is performed in a portion where the lower lid 4 covers the chip body 2. For this reason, the lower lid 4 is preferably made of a metal sheet or a material containing metal. In this case, it is not essential to provide the metal sheet or the metal-containing material on the entire surface of the lower lid 4, and it can be provided only on the portion to be heated.
On the other hand, in the present embodiment, heating is not required in the portion covering the chip body 2 with the upper lid 3. For this reason, the upper lid 3 can be composed of only a less expensive resin film.

Further, the material and shape of the upper lid 3 and the lower lid 4 can be selected in consideration of thermal deformation of the sample analysis chip 1.
If a part of the sample analysis chip 1 is heated to promote the biochemical reaction, the sample analysis chip 1 may be warped or distorted. However, in the present embodiment, the upper lid 3 and the lower lid 4 are attached to the first surface 2f and the second surface 2g of the chip body 2, respectively. Therefore, the curvature and distortion of the sample analysis chip 1 can be reduced by appropriately setting the rigidity and thermal deformation characteristics of each member.

In the present embodiment, as an example, both the upper lid 3 and the lower lid 4 use a bonding material (thickness 150 μm) of aluminum and polypropylene as a material.
In this embodiment, since the sample analysis chip 1 is heated via the lower lid 4, the outer diameter of the lower lid 4 is transmitted so that heat from the lower lid 4 is transmitted only to a necessary range in the sample analysis chip 1. Is smaller than the upper lid 3.

Next, the flow path structure of the chip body 2 will be described.
As shown in FIGS. 3 to 5, an injection hole 2a is provided at the center of the outer shape of the chip body 2 so as to penetrate the injection hole 3a of the upper lid 3 in the thickness direction. The inner diameter of the injection hole 2a may be the same as or different from the inner diameter of the injection hole 3a. In the present embodiment, as an example, the inner diameter of the injection hole 2a and the inner diameter of the injection port 3a are the same.
As shown in FIG. 3, a first reaction chamber 10, a second reaction chamber 12, and an air passage 11 are formed on the second surface 2 g.
As shown in FIG. 4, a measuring chamber 13 is formed on the first surface 2f.

As shown in FIG. 3, the first reaction chamber 10 is an elongated groove formed on the second surface 2g in order to store the sample liquid and cause the first reaction to occur in the entire sample liquid.
The shape of the first reaction chamber 10 is not particularly limited, but in the present embodiment, a circle C1 that is a virtual concentric circle centering on the injection hole 2a and the storage volume per unit area can be increased. Are provided in an annular region sandwiched between circles C2 having a larger diameter than the circle C1, and have a shape meandering in the circumferential direction (circumferential direction of the annular region). In the following description, it is assumed that the circle C1 is an inscribed circle of the region where the first reaction chamber 10 is formed, and the circle C2 is an circumscribed circle of the region where the first reaction chamber 10 is formed. .

In the following, in the description of the shape and arrangement, when a circle related to the shape or arrangement to be described is specified, the direction along the diameter of the circle is the radial direction regardless of whether the circle is a substance or a virtual circle. Called. Regarding the relative position in the radial direction, a position closer to the center of the circle (position close to the center of the circle) is referred to as the radially inner side, and a position closer to the outer periphery of the circle (position closer to the outer periphery of the circle) is referred to as the radially outer side. . Further, in the circle, a direction that circulates perpendicularly to the radial direction is referred to as a circumferential direction.
In addition, in order to simplify description, description of the circle which prescribes | regulates a radial direction may be abbreviate | omitted when there is no possibility of misunderstanding.

The starting end portion 10a of the first reaction chamber 10 is a linear groove extending from the circle C1 to the circle C2 along the radial direction thereof. Further, the starting end portion 10a of the first reaction chamber 10 communicates with an introduction path 2h extending radially outward (from the center of the circles C1 and C2 toward the outer periphery) along the radial direction from the injection hole 2a. Yes.
As shown in FIG. 5, the depth of the start end portion 10a and the introduction path 2h is the same as the depth h10. The depth h10 has a relationship of h10 <h2, for example, when the thickness of the chip body 2 is defined as h2.
As shown in FIG. 3, the terminal portion 10b of the first reaction chamber 10 is formed at a position adjacent to the starting end portion 10a in the circumferential direction. Further, the terminal portion 10b of the first reaction chamber 10 is provided between the vertical hole 2c provided to penetrate the chip body 2 in the thickness direction inside the circle C1 and the circle C2. Furthermore, the terminal portion 10b of the first reaction chamber 10 is a linear groove portion that is provided so as to pass through the center of the vertical hole 2c and to be inclined with respect to the diameter of the circle C2.
As shown in FIG. 6, the terminal portion 10b communicates with the vertical hole 2c. Moreover, the depth from the 2nd surface 2g to the termination | terminus part 10b is the depth h10 like the start end part 10a.

As shown in FIG. 7, an intermediate portion of the first reaction chamber 10 sandwiched between the start end portion 10a and the end end portion 10b is a folded flow passage 10A, a first straight flow passage, clockwise around the injection hole 2a. 10B, the return channel 10C, and the second straight channel 10D have a meandering shape that repeats in this order.
Note that the groove depths of the folded channel 10A, the first straight channel 10B, the folded channel 10C, and the second straight channel 10D are all the depth h10.

In the folded flow path 10A, the flow path from the radially inner side (position closer to the circle C1) to the radially outer side (position closer to the circle C2) is folded back approximately 180 degrees (including the case of 180 degrees) to the radially inner side. It is the flow path which has U shape provided so that it might return.
The first straight flow path 10B is connected to the folded flow path 10A, and is a flow path that is directed radially inward along a direction inclined with respect to the radial direction of the circle C2 (a direction toward a position shifted from the center of the circle C2). It is. The groove width of the first straight channel 10B decreases as it goes inward in the radial direction.
The inclination direction of the first straight channel 10B is not particularly limited as long as it is inclined in a certain direction. In the present embodiment, as an example, as illustrated in FIG. 3, the first linear flow path 10 </ b> B is inclined so as to proceed in the clockwise direction from the radially outer side toward the radially inner side.

The folded flow path 10C is a flow path having a U-shape provided so that the first straight flow path 10B is folded by approximately 180 degrees (including the case of 180 degrees) and folded to the circle C2 side.
The groove width of the return channel 10C is substantially the same as the groove width at the connection portion between the return channel 10C and the first straight channel 10B, and is narrower than the groove width of the return channel 10A.
The second straight channel 10D is a straight channel that is inclined in parallel with the first straight channel 10B folded back by the folded channel 10C and is parallel to the circle C2. The groove width of the second straight channel 10D increases as it goes outward in the radial direction.
The radially outer end of the second straight channel 10D is connected to another adjacent folded channel 10A.

In the intermediate part of the first reaction chamber 10, such a meandering flow path structure is repeated, so that the first straight flow path 10B and the second straight flow path 10D have substantially the same width (including the case of the same width). ) In parallel with a gap. That is, the first straight flow path 10B and the second straight flow path 10D are partitioned by the

convex portions

2j and 2i having a substantially constant wall thickness and a height h10.
The wall thicknesses of the

convex portions

2j and 2i are the strength necessary for pumping the sample liquid in the first reaction chamber 10, the adhesive strength of the lower lid 4 on the

convex portions

2j and 2i, In consideration of workability, it can be set to an appropriate size.
Within the range of these conditions, if the wall thickness of the

convex portions

2j and 2i is reduced, the area of the first reaction chamber 10 in the regions of the circles C1 and C2 can be maximized.
In the present embodiment, the

convex portions

2j and 2i are provided so as to extend obliquely with respect to the radial direction. Thereby, the change of a channel width | variety can be suppressed, enlarging the area of the 1st linear flow path 10B and the 2nd linear flow path 10D. By suppressing the change in the channel width, the flow of the sample liquid in the meandering first reaction chamber 10 becomes good.
When the

convex portions

2j and 2i are provided so as to extend along the radial direction, and the areas of the first linear flow channel 10B and the second linear flow channel 10D are made to have the same size, the radially inner side in each flow channel The difference between the channel width and the outer channel width becomes larger. For this reason, the flow of the sample liquid in the meandering first reaction chamber 10 is deteriorated, and the sample liquid may not be satisfactorily filled.

As shown in FIG. 3, the second reaction chamber 12 introduces the sample liquid (first reaction liquid) weighed by the measuring chamber 13 described later, and the second reaction chamber 12 individually receives the second sample liquid from the weighed sample liquid. It is a groove for causing a reaction.
On the second surface 2g outside the region where the first reaction chamber 10 is provided (outside the circle C2), a plurality of the second reaction chambers 12 are arranged along the circumferential direction of the circle C2 at an equal pitch (equally spaced). ) (A second reaction chamber group having a plurality of second reaction chambers 12 is provided).
The structures of the second reaction chambers 12 are the same, and as shown in FIG. 7, the shape in plan view has an oval shape extending along the radial direction of the circle C2 with a longitudinal axis. It is a groove.

In order to transfer the sample liquid (first reaction liquid) from the measurement chamber 13 to the radially inner end of each second reaction chamber 12, a transfer path that is a shallower and narrower groove than the second reaction chamber 12. 2k are connected.
As shown in FIG. 8, the depth of the second reaction chamber 12 is h12 (where h12 <h2), and the depth of the transfer path 2k is h2k (where h2k <h12).
The area and depth h12 in plan view of the second reaction chamber 12 are appropriately set based on the volume of the sample liquid (first reaction liquid) necessary for performing the second reaction.

As shown in FIG. 7, the transfer path 2 k extends from the radially inner end of the second reaction chamber 12 along the radial direction, and then extends in an oblique direction so as to intersect the radial direction. . Further, in the transfer path 2 k, the radially inner end communicates with the vertical hole 2 e that penetrates in the thickness direction of the chip body 2.

A branch path 2m having a common groove depth with the transfer path 2k is formed on the side opposite to the extending direction on the radially inner side of the transfer path 2k from the bent portion in the intermediate portion of the transfer path 2k.
The branch path 2m extends substantially along the circumferential direction of the circle C2, and communicates with the excess liquid storage chamber 14 disposed adjacent to the second reaction chamber 12 in the circumferential direction.

The surplus liquid storage chamber 14 is a groove formed in the second surface 2 g in order to store surplus liquid overflowing from the second reaction chamber 12.
Examples of the excess liquid include a sample liquid and a liquid introduced into the second reaction chamber 12 in addition to a sample liquid such as a lid solution described later.
The shape of the surplus liquid storage chamber 14 is set so that a volume capable of storing all the surplus liquid is obtained in consideration of variation in the amount of liquid such as the sample liquid (first reaction liquid) transferred from the measuring chamber 13. .
In the present embodiment, as an example, the shape of the surplus liquid storage chamber 14 in plan view is a radial shape that extends along the radial direction of the circle C2 and whose circumferential width increases from the radially inner side to the outer side. Or it is a groove part formed in pear shape. The depth of the excess liquid storage chamber 14 is, for example, h12.

As shown in FIG. 3, the air passage 11 is formed on the second surface 2 g inside the circle C <b> 1 in order to constitute a flow path for discharging fluid such as air inside the measuring chamber 13 described later to the outside. It is a groove.
In the air passage 11, an end portion (first end portion) provided in the vicinity of the circle C <b> 1 communicates with a vertical hole 2 d penetrating in the thickness direction of the chip body 2.
The other end (second end) of the air passage 11 opens at a position overlapping the air hole 3b of the upper lid 3 and communicates with the vertical hole 2b penetrating in the thickness direction of the chip body 2.
The shape of the air passage 11 in plan view is not particularly limited as long as an independent flow path can be formed between the

vertical holes

2d and 2b. In this embodiment, the ventilation path 11 is provided in the side of the introduction path 2h, and is formed in S shape in planar view.
As shown in FIG. 9, the depth of the ventilation path 11 is h11 (however, h11 <h2). The depth h11 of the ventilation path 11 only needs to have a depth that allows the fluid in the measurement chamber 13 to be smoothly discharged.

It is possible to introduce not only excess air but also excess liquid introduced into the measuring chamber 13 and overflowing from the measuring chamber 13, such as a sample solution and a lid solution described later, into the air passage 11. is there. Therefore, the air passage 11 is a surplus liquid storage chamber that stores surplus liquid overflowing from the measuring chamber 13, and includes the first reaction chamber 10, the measuring chamber 13, and the second reaction chamber 12 (second reaction chamber group). ) Is formed in a surplus liquid storage chamber provided in a central region (inside the small diameter circle of the circles C1 and C3).

As shown in FIG. 4, the measurement chamber 13 measures the sample liquid (first reaction liquid) in which the first reaction has occurred, and uses the centrifugal force to measure the sample liquid (first reaction liquid) that has been weighed. These are grooves formed on the first surface 2 f for moving to the respective second reaction chambers 12.
The measuring chamber 13 stores the sample liquid (first reaction liquid) in a measuring flow path 13b for measurement, and the sample liquid stored in the measurement flow path 13b (first reaction liquid) in the radial direction by centrifugal force. And a plurality of branch paths 13F for taking out to the outside.
The groove depth of the measuring chamber 13 is constant at the depth h13 (where h13 <h2).

The measurement flow path 13b meanders in the circumferential direction in an annular region sandwiched between a circle C3, which is a virtual concentric circle centered on the injection hole 2a, and a circle C4 having a larger diameter than the circle C3. It is composed of an elongated channel. In the following description, it is assumed that the circle C3 is an inscribed circle of the area where the measuring flow path 13b is formed, and the circle C4 is a circumscribed circle of the area where the measuring flow path 13b is formed.
In the present embodiment, the diameter of the circle C3 is smaller than the diameter of the circle C1. The diameter of the circle C4 is larger than the diameter of the circle C3 and smaller than the diameter of the circle C2.
The start end portion 13a of the measurement channel 13b communicates with the vertical hole 2c.
The end portion 13c of the measurement channel 13b is disposed at a position adjacent to the start end portion 13a in the circumferential direction, and communicates with the vertical hole 2d.
For this reason, as shown in FIG. 4, the measurement flow path 13b flows in a circular region sandwiched between the circles C3 and C4 from the start end portion 13a substantially once in the counterclockwise direction to the end portion 13c. Forming a road.
With such an arrangement, the metering channel 13b is located in a region where a part of the metering channel 13b overlaps the first reaction chamber 10 in the thickness direction of the chip body 2.

As shown in FIG. 10, the metering flow path 13b includes the first straight flow path 13A, the outer peripheral bent flow path 13B, the second straight flow path 13C, and the inner peripheral side in the counterclockwise direction centering on the injection hole 2a. The bent channel 13D has a meandering shape formed repeatedly in this order.
In addition, the start end part 13a of the measurement flow path 13b is the first straight flow path 13A, and the end part 13c is the inner circumferential side bent flow path 13D.

13 A of 1st straight flow paths, the outer periphery side bending flow path 13B, and the 2nd straight flow path 13C comprise the U-shaped flow path as a whole, and while U-shaped opening part faces the circle C3, U An outer peripheral side bent flow path 13B, which is a letter-shaped bent portion, is arranged so as to be inscribed in the circle C4.
As will be described later, the sample liquid (first reaction liquid) stored in the first straight flow path 13A, the outer peripheral bent flow path 13B, and the second straight flow path 13C is collected in one second reaction chamber 12. Moved. For this reason, the volumes of the first straight flow path 13A, the outer peripheral bent flow path 13B, and the second straight flow path 13C are appropriately set according to the amount of sample liquid.
The first straight flow path 13A and the second straight flow path 13C have a diameter L of a circle C4 passing through the bending center of the outer peripheral bent flow path 13B and angles θ1, θ2 (where θ1, θ2 are acute angles), respectively. So that they cross. The angles θ1 and θ2 may have different sizes and inclination directions.
The angles θ1 and θ2 are preferably 0 ° or more and 60 ° or less, respectively.
In addition, in order to increase the ratio of the measurement amount to the area of the annular region sandwiched between the circles C3 and C4 and to keep the measurement chamber 13 in a compact size, the angles θ1 and θ2 are made to coincide with each other. It is preferable to align the inclination direction. In this case, as the angles θ1 and θ2 are increased within the above preferable range (0 ° to 60 °), the annular region can be narrowed.
In the present embodiment, as an example, θ1 = θ2 = 25 (°) is set. As shown in FIG. 10, the inclination direction is a direction that moves counterclockwise around the injection hole 2a along the first linear flow path 13A from the radially inner side toward the radially outer side.

The diameter of the groove width of the first straight channel 13A increases from the radially inner side toward the radially outer side.
The groove width of the second straight channel 13C is constant, for example, the same width as the minimum groove width of the first straight channel 13A.
As described above, the outer peripheral bent flow path 13B is a flow path having an arc shape that smoothly connects the first straight flow path 13A and the second straight flow path 13C having different groove widths on the outer side in the radial direction.
The outer peripheral bending channel 13B has the largest groove width at the outermost peripheral portion in the radial direction.

The inner circumferential side curved channel 13D includes the first linear channel 13A, the U-shaped outer circumferential side curved channel 13B, and the second linear channel 13C, which is an adjacent channel among the second linear channel 13C. It is a groove part which makes the edge part inside radial direction with 13 A of 1st linear flow paths communicate in circular arc shape.
The groove width of the inner circumferential side bent flow path 13D is about the same as that of the second straight flow path 13C.

The branch path 13F is an elongated groove extending outward in the radial direction from a portion of the outer peripheral bent flow path 13B constituting the outermost peripheral portion of the measurement flow path 13b.
The groove width of the branch path 13F is preferably the same as that of the transfer path 2k, for example.
In this embodiment, after extending along the substantially radial direction from the outer periphery side bending flow path 13B, it is inclined and extended in the direction opposite to the inclination direction of the first linear flow path 13A.
As shown in FIG. 11, the end of each branch path 13F in the extending direction communicates with a vertical hole 2e to which one transfer path 2k is connected. Therefore, the branch path 13F communicates with the second reaction chamber 12 on a one-to-one basis via the transfer path 2k and the vertical hole 2e.
The branch path 13F and the transfer path 2k communicated with each other through the vertical hole 2e may have the same or different extending directions. In the present embodiment, as an example, an example in which the branch path 13F and the transfer path 2k indicate the same extending direction is used. For this reason, the branch path 13F and the transfer path 2k are aligned in the same straight line in plan view.

With such a configuration, a plurality of channel structures are formed in the sample analysis chip 1.
For example, when a fluid is injected from the injection port 3a, the fluid descends to the second surface 2g side through the injection hole 2a, enters the start end portion 10a of the first reaction chamber 10 via the introduction path 2h, One end of the reaction chamber 10 is reached through the middle portion of the reaction chamber 10.
The fluid that has reached the end portion 10 b rises to the first surface 2 f side through the vertical hole 2 c and reaches the start end portion 13 a of the measuring chamber 13.

In the measuring chamber 13, a branch path 13 </ b> F is formed on the side of the measuring channel 13 b, and the terminal portion 13 c of the measuring chamber 13 is lowered to the second surface 2 g side through the vertical hole 2 d and enters the vent path 11. An inward flow path (hereinafter referred to as “discharge flow path”) is formed.
The flow path following each branch path 13F descends to the second surface 2g side through the vertical hole 2e, and the flow path toward the second reaction chamber 12 through the transfer path 2k and the branch path from the middle of the transfer path 2k. It divides into 2 m and is divided into a flow path toward the surplus liquid storage chamber 14.
Since neither the second reaction chamber 12 nor the surplus liquid storage chamber 14 is connected to a flow path communicating with the outside, air is confined in an unused state. For this reason, in order to move the fluid from the branch path 13F to the transfer path 2k, it is necessary to apply at least a corresponding external force that resists the air pressure to the fluid.

Since the air passage 11 communicates with the vent hole 3b through the exhaust passage, the fluid in the metering passage 13b is discharged from the metering passage 13b rather than going to the second reaction chamber 12 through the branch passage 13F. It is much easier to flow to the ventilation path 11 through the flow path.
In the case of a gas such as air, the fluid that has reached the air passage 11 is sequentially discharged upward through the air holes 3b. Further, when the fluid reaching the air passage 11 is a liquid, the air enters the air passage 11 while discharging the air in the air passage 11 from the air vent 3b, and the volume range of the air passage 11 is limited. Is stored in the air passage 11.
As described above, since the vent hole 3b is provided in the upper part of the sample analysis chip 1, the clogging of the vent hole 3b due to the ingress of liquid is prevented, and the liquid is not easily leaked to the outside.

Such a sample analysis chip 1 can be manufactured by manufacturing the chip body 2, the upper lid 3, and the lower lid 4 and bonding them together.
Before performing this bonding, a reaction reagent for performing the second reaction is fixed in each second reaction chamber 12.
In each of the second reaction chambers 12, the same reagent may be used. However, depending on the purpose of analysis, a plurality of types of second reactions may be performed, and thus different reagents may be used.
When different reagents are fixed in each second reaction chamber 12, a plurality of processes can be performed on one type of specimen (sample).
Alternatively, a part of the reagent for performing the reaction may be fixed to each second reaction chamber 12 and the remaining reagent may be introduced together with the sample solution.
Similarly, the reagent used for causing the first reaction in the first reaction chamber 10 may be fixed in the first reaction chamber 10 or introduced together with the sample solution.

As an example of the reagent fixing method, for example, a liquid reagent is dropped into the first reaction chamber 10 and the second reaction chamber 12 of the chip body 2 with a pipette or the like, and the chip body 2 is centrifuged at 2000 rpm to 3000 rpm for about 5 minutes. A method of rotating and then drying can be used.
In this case, an appropriate amount of the liquid reagent remains in a flat state by the action of centrifugal force due to rotation, and the liquid reagent is dried, so that the first reaction chamber 10 and the second reaction chamber 12 The reagent can be fixed so that the layer thickness is constant.

As another method for fixing the reagent, a method using a hot-melt material is also possible.
For example, the hot-melt material is disposed so as to cover the reagent by dropping a molten hot-melt material onto the reagent disposed in the first reaction chamber 10 and the second reaction chamber 12 of the chip body 2. When the hot-melt material is solidified, the reagent is fixed.

The heat-meltable substance used in this fixation method should be solidified and liquefied at the temperature used for the analysis, should not adversely affect the reaction and liquid transfer, and can be replaced with the sample liquid. Any appropriate material that satisfies the above conditions can be used.
For example, when performing a biochemical reaction such as PCR (polymerase chain reaction), a wax dedicated to the biochemical reaction can be used.
For example, paraffin wax can be used. In this case, in order to change the melting temperature, a plurality of waxes having different melting points can be mixed and used.

As a method of attaching the upper lid 3 and the lower lid 4 to the chip body 2, in addition to a method using an adhesive or the like, a resin coating layer is provided as an adhesive layer on one surface of members to be bonded to each other, and the resin coating layer is A method of adhering members by melting them is mentioned.
In the case of providing a resin coating layer, it is preferable to provide a resin coating layer on a member having high thermal conductivity and melt-bond it. For example, if the member to be bonded is a member including a metal material and a resin material, it is preferable to provide a resin coating layer on the member including the metal material.
Examples of the material for the resin coating layer include synthetic resins such as PET (polyethylene terephthalate) resin, polyacetal resin, polyester resin, and polypropylene resin.

Further, when the resin coating layer is formed, welding using a laser is possible by forming an anchor layer as a base of the resin coating layer.
For example, carbon black (light-absorbing material) that absorbs laser wavelength light is kneaded in the anchor layer, and the resin coating layer can be melted and bonded by irradiating laser light.
Alternatively, instead of adding carbon black to the anchor layer, carbon black may be added to the resin coating layer, or the surface of the resin coating layer may be painted black. For example, the resin coating layer can be efficiently melted by irradiating infrared laser light having a wavelength of about 900 nm.
Laser welding, unlike thermal welding, does not require heating the chip body 2 in a wide range, so that the base body 2 and the reagent fixed to the chip body 2 can be attached without affecting the heating. Can be combined.

Next, the operation of the sample analysis chip 1 in biochemical analysis will be described focusing on the sample analysis method using the sample analysis chip 1.
The sample analysis chip 1 according to this embodiment can be used for detection and analysis of biochemical substances in samples such as DNA and protein.
In order to detect and analyze these biochemical substances using the sample analysis chip 1, a lid solution is poured from the inlet 3a in order to prevent evaporation and contamination (hereinafter referred to as contamination) of the sample liquid.
At this time, the lid solution is preferably a stable substance such as mineral oil or silicone oil, and does not inhibit the first reaction described later and has a specific gravity smaller than that of the sample liquid.

The sample analysis apparatus that performs analysis using the sample analysis chip 1 has, as a mechanism for sample analysis, for example, a dispensing mechanism that injects a fluid such as a reagent, sample liquid, and air described later into the injection port 3a, and the sample analysis chip 1. A heating mechanism for heating the sample analysis chip 1, a rotation mechanism for rotating the sample analysis chip 1, and a detection mechanism for detecting a biochemical reaction in the sample analysis chip 1 or a measurement mechanism for measuring the biochemical reaction.
The detection mechanism or the measurement mechanism may be arranged anywhere around the sample analysis chip 1 or on the surface as long as it can detect and measure a biochemical reaction. For example, it can be arranged at a position facing each second reaction chamber 12 through the lower lid 4 of the sample analysis chip 1 or on the side surface of the chip body 2.
When using the detection mechanism or the measurement mechanism, the position of the sample analysis chip 1 is rotated by the rotation mechanism, thereby moving the target part to be detected and measured with respect to the detection mechanism or the measurement mechanism. Can be done.
All of these mechanisms for sample analysis may be included in one apparatus, or one or more mechanisms may be constituted by another apparatus or another instrument.

Next, the sample liquid to be analyzed is injected from the injection port 3a using a dispensing mechanism or a dispensing instrument. The sample liquid is injected into the first reaction chamber 10 via the injection hole 2a and the introduction path 2h.
When an amount of sample liquid necessary for analysis is injected into the first reaction chamber 10, a lid solution for preventing evaporation and contamination of the sample liquid is poured again from the injection port 3a.

Next, in the first reaction chamber 10, the sample solution and the reagent disposed in the first reaction chamber 10 are mixed to cause the first reaction. However, the first reaction occurs only by mixing the sample solution and the reagent. Absent. For example, when a biochemical reaction such as a PCR reaction is performed as the first reaction, heat treatment or the like needs to be performed on the sample solution in the first reaction chamber 10.
Examples of the heating mechanism and the heating tool for performing the heat treatment include a mechanism and a tool using a cartridge heater, a heating wire, a Peltier element, and the like.
For example, the sample liquid in the first reaction chamber 10 covered by the lower lid 4 is heat-treated by heating the lower lid 4 with a cartridge heater. Thereby, in the 1st reaction chamber 10, the 1st reaction which requires heat treatment like PCR reaction occurs, for example.
For example, in the PCR reaction, when the sample DNA increases sufficiently, the heating is stopped and the first reaction is terminated.
Thus, the 1st reaction in this embodiment is a biochemical reaction performed with respect to the whole sample liquid.

Next, the sample liquid (first reaction liquid) subjected to the first reaction is moved to the measuring chamber 13.
In the present embodiment, the sample liquid (first reaction liquid) in the first reaction chamber 10 is raised to the first surface 2 f side via the vertical hole 2 c and moved into the measurement flow path 13 b of the measurement chamber 13. .
This movement is performed by, for example, injecting air or oil from the injection port 3a using a pipette, a syringe, or the like, so that the sample liquid (first reaction liquid) in the first reaction chamber 10 is discharged from the vertical hole 2c. Do by pushing up.
That is, by injecting air, oil, or the like corresponding to the volume of the first reaction chamber 10 from the injection port 3a, the sample liquid (first reaction solution) in the first reaction chamber 10 is injected into the measuring chamber 13. Can be moved sequentially.
At this time, the pressure for injecting air, oil, or the like is such that the sample liquid (first reaction liquid) does not flow into the branch path 13F. That is, the air pressure and the sample liquid (which are hereinafter referred to as “measuring channel side space”) held in the space of the branch path 13F, the transfer path 2k, the second reaction chamber 12, the branch path 2m, and the excess liquid storage chamber 14 The pressure should not exceed a resistance pressure derived from the surface tension of the first reaction solution.
For this reason, for example, by setting the size of the vent hole 3b to an appropriate size, the resistance of the discharged fluid in the discharge channel of the measuring chamber 13 is made sufficiently lower than the resistance pressure. .

In this way, the sample liquid (first reaction liquid) moved to the measuring chamber 13 through the vertical hole 2c advances through the measuring channel 13b without entering the side space of the measuring channel.
The fluid existing in the metering channel 13b before the introduction of the sample solution (first reaction solution) is sequentially pushed out to the metering channel 13b side by the sample solution (first reaction solution), and aerated through the discharge channel. It moves to the path 11 and is discharged to the outside from the vent hole 3b.
In this way, when the sample liquid (first reaction liquid) in the first reaction chamber 10 is moved to the measuring chamber 13 by a necessary amount, the injection of air or oil is terminated.

Next, the sample analysis chip 1 is rotated around a rotation axis extending in the normal direction of the chip body 2 through the centers of the circles C3 and C4 in which the measurement chambers 13 are arranged, so that a sample liquid (first reaction liquid) is obtained. Weigh the amount. In the present embodiment, the centers of the circles C3 and C4, the center of the outer shape of the chip body 2, and the central axis of the injection port 3a are all coaxial.
The sample analysis chip 1 can be rotated using a centrifuge or the like having a rotation mechanism that holds the sample analysis chip 1 and rotates the sample analysis chip 1 around the central axis of the injection port 3a.
As for the rotation speed of the rotation mechanism, the centrifugal force acting on the sample liquid (first reaction liquid) in the measurement flow path 13b generates the air pressure in the side space of the measurement flow path and the surface tension of the sample liquid (first reaction liquid) itself. The rotational speed that flows into the second reaction chamber 12 at least is required.
Such a rotational speed depends on the shape and dimensions of the flow channel structure, but for example, when the average radius of the measurement flow channel 13b is 15 mm, a sufficient centrifugal force can be obtained if it is about 1000 rpm or more. .

When the sample analysis chip 1 is rotated at such a rotation speed, the sample liquid (first liquid crystal) located in the first straight flow path 13A, the outer circumferential bent flow path 13B, and the second straight flow path 13C in the measurement flow path 13b. The reaction solution) moves to the radially outer branch 13F by the action of centrifugal force.
The sample liquid (first reaction solution) that fills each inner peripheral bent channel 13D is adjacent to the first straight channel 13A and the inner periphery adjacent to the top of the bend of the inner peripheral bent channel 13D in contact with the circle C3. It is divided into any of the side bent flow paths 13D and moves radially outward.
For this reason, the sample liquid is included in the branch path 13F within a U-shaped space formed by the first straight flow path 13A, the outer peripheral bent flow path 13B, and the second straight flow path 13C communicating with the branch path 13F. (First reaction liquid) flows in. As a result, the sample liquid (first reaction liquid) moves toward the second reaction chamber 12 in a state where the sample liquid (first reaction liquid) is weighed according to the volume of the U-shaped space.

The sample liquid (first reaction liquid) moved to the branch path 13F moves down the vertical hole 2e and moves to the transfer path 2k.
Although the transfer path 2k is branched into the branch path 2m, since the centrifugal force acts along the radial direction, the sample liquid (first reaction liquid) does not flow into the branch path 2m along the circumferential direction. Then, it moves through the transfer path 2k and moves to the second reaction chamber 12.
When the sample liquid (first reaction liquid) exceeding the volume of the second reaction chamber 12 tries to flow in, the sample liquid (first reaction liquid) overflowing from the second reaction chamber 12 branches to the branch path 2m, and surplus It moves to the liquid storage chamber 14.
For this reason, a sample liquid (first reaction liquid) equal to the volume of the second reaction chamber 12 remains in the second reaction chamber 12.
Thereby, in this embodiment, even if variation occurs in the volume of the sample liquid (first reaction liquid) measured in each U-shaped space of the measurement flow path 13b, the excess sample liquid (first reaction) By discharging the liquid) to the excess liquid storage chamber 14, the sample liquid (first reaction liquid) can be measured more accurately according to the volume of the second reaction chamber 12.
In this way, when all of the sample liquid (first reaction liquid) in the measurement channel 13b has moved to the side space of the measurement channel, the rotation of the sample analysis chip 1 is stopped.

Next, a biochemical reaction as a second reaction is performed using the sample liquid (first reaction liquid) sent to the second reaction chamber 12.
That is, the second reaction corresponding to each reagent is performed by mixing the reagent arranged in each second reaction chamber 12 and the sent sample liquid (first reaction liquid) (the second reaction liquid is changed). can get).
The reaction state of the second reaction can be detected and measured by a detection mechanism or a measurement mechanism corresponding to the type of the second reaction. For example, the reaction can be detected by a technique such as fluorescence detection.
Thereby, a sample can be analyzed based on the detected or measured reaction state.

As described above, according to the sample analysis chip 1 according to the present embodiment, the first reaction chamber 10 includes the first reaction chamber 10 and the plurality of second reaction chambers 12 (second reaction chamber group). And a plurality of types of biochemical reactions can be performed by the sample liquid moved to each of the plurality of second reaction chambers 12.
At this time, since the measuring chamber 13 is provided between the first reaction chamber 10 and the second reaction chamber 12, the sample liquid (first reaction liquid) moved to the measuring chamber 13 is measured using centrifugal force. be able to. For this reason, since the weighed sample liquid (first reaction liquid) can be immediately moved to the second reaction chamber 12 to perform the second reaction, unevenness in the amount of sample liquid to be reacted is reduced (sample liquid volume). The sample can be analyzed with high accuracy and speed.
Further, in this embodiment, the first reaction chamber 10 and the second reaction chamber 12 (second reaction chamber group) are on the second surface 2g side, and the weighing chamber 13 is on the first surface 2f side. The first reaction chamber 10 and the measuring chamber 13 are partially spaced from each other in the thickness direction of the chip body 2.
For this reason, space saving is possible, and a compact configuration in which the outer diameter of the sample analysis chip 1 is reduced as compared with the amount of sample liquid accommodated is possible.

Since the measurement chamber 13 and the first reaction chamber 10 are spaced apart from each other in the thickness direction, the measurement chamber 13 and the first reaction chamber 10 that do not require thermal reaction can be thermally separated from each other. It becomes possible. For this reason, even if the first reaction chamber 10 is heated, a structure in which unnecessary heat is hardly transmitted to the measuring chamber 13 is possible.

The measuring chamber 13 and the second reaction chamber 12 (second reaction chamber group) are arranged such that the measuring chamber 13 is disposed above and the second reaction chamber 12 (second reaction chamber group) is disposed below, and the plate is disposed through the vertical hole 2e. It communicates in the thickness direction.
Therefore, in the second reaction chamber 12, bubbles generated at the time of the heating reaction or chemical reaction can be easily released upward from the second reaction chamber 12, or a lid solution such as oil sent to prevent evaporation and contamination. It is possible to easily separate the reaction solution in the thickness direction.

The first reaction chamber 10 and the second reaction chamber 12 (second reaction chamber group) are formed in concentric annular regions adjacent to each other in the radial direction and covered with the lower lid 4. For this reason, when the heating by the lower lid 4 is performed for the first reaction, the second reaction chamber 12 can also be heated at the same time, so that the second reaction chamber 12 can be preheated. Thereby, the second reaction can be performed in a short time after the sample liquid (first reaction liquid) has moved to the second reaction chamber 12.

In the sample analysis chip 1, the injection port 3a, the injection hole 2a, the ventilation path 11, and the ventilation hole 3b are surrounded by the first reaction chamber 10, the measurement chamber 13, and the second reaction chamber 12 (second reaction chamber group). In the central area.
That is, the inlet 3a, which is an opening to the outside air at a position closer to the center point of the centrifugal axis than the flow path of the first reaction chamber 10, the measurement chamber 13, and the second reaction chamber 12 (second reaction chamber group), There is a vent 3b. For this reason, when the sample analysis chip 1 is centrifugally rotated, the backflow of the oil or sample liquid from the flow path to the inlet 3a and the vent hole 3b is prevented, and excess air and bubbles are easily removed from the substrate. Can do.

As described above, according to the sample analysis chip 1, the first surface and the second surface of the plate-like part are separated (separately) from the first reaction chamber, the measurement chamber, and the second reaction chamber (second Since the reaction chamber group is provided, it is easy to send and measure the sample liquid to be analyzed, and a plurality of types of biochemical reactions can be performed even with a compact configuration.
For example, the processing for the whole and the processing for the divided state can be performed in stages on one type of specimen. Therefore, after first performing amplification, storage, measurement, etc. of a valuable DNA sample contained in the sample solution, different processing can be performed on the sample solution distributed by the measurement chamber.
Moreover, according to the sample analysis chip 1, when the first stage process (first reaction) is performed on the sample solution, a solution serving as a lid that can prevent evaporation and contamination can be placed. . In this case, if the solution is adjusted so that the specific gravity of the lid solution is lighter than the specific gravity of the sample liquid, the centrifugal force is applied after the first reaction, and the sample liquid (first reaction) is caused by the difference in specific gravity. Liquid), the lid solution is separated into the upper layer. As a result, the lid solution continues to act as a lid even after the second reaction, and contamination and evaporation between the sample solutions can be prevented.

Next, an example of sample analysis in which the sample analysis chip 1 according to this embodiment can be used will be described.
Examples of sample analysis in gene analysis include detection of somatic mutations and detection of germline mutations. Since the type of protein to be expressed differs depending on the genotype, for example, it causes a difference in the function of the metabolic enzyme of the drug, resulting in individual differences in the optimal dose of the drug and the likelihood of side effects. By making use of this in the medical field and examining the “genotype” of each patient, it is possible to provide tailor-made medical care.

-Detection of SNPs About 0.1% of the human genome has a unique nucleotide sequence difference, which is called SNP (Single Nucleotide Polymorphism), and is one of germline mutations. As one of the SNP identification methods, for example, a PCR-PHFA (PCR-Preferences Modulation Formation Assay) method using fluorescence is used. The PCR-PHFA method includes a PCR process for amplifying a detection mutation site and a competitive strand displacement reaction process using an amplified fragment and a corresponding probe.
According to this method, in the second reaction, a mutation is detected by the difference in luminescence of the fluorescent reagent, but by using the sample analysis chip 1 according to the present embodiment, there is little variation in liquid distribution in each second reaction chamber 12. Therefore, accurate SNP detection can be performed.
Similarly, the sample analysis chip 1 according to this embodiment can be used for the Invader (registered trademark) method, the Taqman PCR method and the like as SNP detection methods other than those described above.

As another example of sample analysis, PCR for SNPs involved in side effects on warfarin (anti-coagulant, used as a drug for heart disease and hypertension) using the sample analysis chip 1 according to the present embodiment will be described below. -An analysis example using the PHFA method will be described.

A sample nucleic acid obtained from blood or the like is purified to obtain a solution sample. After injecting this solution sample into the sample analysis chip 1 as a sample solution, amplification of the sample nucleic acid (first reaction) is performed by PCR in the first reaction chamber 10 (a first reaction solution is obtained).
Note that SNPs in VKORC1 and CYP2C9 are often discussed for detection of SNPs involved in warfarin, and CYP2C9 * 2 and CYP2C9 * 3 are famous. Gene fragments containing these SNPs from the specimen are amplified by multiplex PCR.

In the detection method described above, two detection wells are required to determine one SNP. Therefore, the sample analysis chip 1 in which 10 or more second reaction chambers 12 are formed is used for each sample. It is preferable. A reagent for SNP detection for performing competitive strand displacement reaction is fixed in each second reaction chamber 12.

The sample (first reaction solution) obtained by amplifying the nucleic acid by the PCR is weighed in the weighing chamber 13 and then filled into each second reaction chamber 12. The temperature of each second reaction chamber 12 is adjusted, and a mutation is detected based on the difference in luminescence of the fluorescent reagent mixed in the reagent fixed in each second reaction chamber 12.
If only one of the two second reaction chambers 12 is positive for one SNP, it can be determined to be homo, and if two are positive, it can be determined to be hetero.

-Detection of K-ras gene mutations Most of the mutations characteristic of upper cancer cells and the resistance to molecular target drugs are somatic mutations. In the case of germ cell mutations (SNP, etc.), a common mutation is seen in all cells, whereas in somatic mutations, cells are mutated only in the mutated cells, and cells that are not mutated (usually normal cells) ) Shows no mutation.

In other words, when many of the sample liquids are normal cells and partially mutated cells are contained, a few mutated genes present in many normal genes must be detected. This is the difference between mutation detection in germ cells and mutation detection in somatic cells, which makes detection of genetic mutations in somatic cells more difficult.

The K-ras gene is a gene whose molecular target drug has been shown to be ineffective in most patient groups when mutations are present in cancer cells. This gene should be detected simply, quickly, inexpensively and with high accuracy. Is being hoped for.

Hereinafter, as another example of sample analysis, an analysis example of the K-ras gene in the PCR-PHFA method will be described.

In the sample analysis chip 1, a reagent containing a probe nucleic acid is fixed in the second reaction chamber 12 for detecting the gene mutation. In detection of the K-ras gene, since there are 13 types of mutations with the wild type, the sample analysis chip 1 in which at least 14 second reaction chambers 12 are formed is used. -It is preferable that a reagent corresponding to detection of the ras gene is immobilized.

Collect cancer cells such as colorectal cancer and purify the sample nucleic acid to obtain a sample solution sample (sample solution). After injecting this sample solution into the sample analysis chip 1, first, a sample nucleic acid is amplified by PCR (first reaction) in the first reaction chamber 10 (a first reaction solution is obtained).

The sample (first reaction solution) obtained by amplifying the nucleic acid by the PCR is weighed in the weighing chamber 13 and then filled in each second reaction chamber 12. By adjusting the temperature of the second reaction chamber 12, the mutation can be detected based on the difference in luminescence of the fluorescent reagent mixed in the reagent fixed in the second reaction chamber 12.

[Second Embodiment]
A sample analysis chip according to the second embodiment of the present invention will be described.
FIG. 12A is a schematic plan view showing the configuration of a sample analysis chip according to the second embodiment of the present invention. FIG. 12B is a schematic front view showing the configuration of the sample analysis chip according to the second embodiment of the present invention. FIG. 13 is a plan view of the sample analysis chip in FIG. 12B viewed from the D direction. FIG. 14 is a schematic back view of the base material used in the sample analysis chip according to the second embodiment of the present invention. FIG. 15 is a schematic plan view of a substrate used for a sample analysis chip according to the second embodiment of the present invention. FIG. 16 is a schematic cross-sectional view of the vent hole of the sample analysis chip according to the second embodiment of the present invention. 17 is a cross-sectional view taken along line EE shown in FIG. 12A.

As shown in FIGS. 12A, 12B, and 13, a sample analysis chip 21 according to the present embodiment is replaced with a chip body 2 instead of the chip body 2 and the lower lid 4 of the sample analysis chip 1 according to the first embodiment. 22 (base material, plate-like portion) and a lower lid 24 (lid material).
Hereinafter, the second embodiment will be described focusing on differences from the first embodiment.

Like the chip body 2 according to the first embodiment, the chip body 22 is a plate member (a plate having a first surface 2f and a second surface 2g sandwiched between a first surface 2f and a second surface 2g. The outer shape in plan view is circular.
The chip body 22 is different from the chip body 2 only in the flow path structure formed on the surface and inside of the chip body 22.

The lower lid 24 is different from the lower lid 4 according to the first embodiment only in that it is a disk-like member that covers the entire second surface 2g of the chip body 22.

Next, the flow path structure of the chip body 22 will be described.
As shown in FIG. 14, the flow path structure on the second surface 2g of the chip body 22 is the same as that of the first embodiment, except that the air passage 11 and the excess liquid storage chamber 14 according to the first embodiment are removed. Instead of the first reaction chamber 10 and the second reaction chamber 12, a first reaction chamber 30 and a second reaction chamber 32 are provided.
As shown in FIG. 15, on the first surface 2f of the chip body 22, a measuring chamber 33 is provided instead of the measuring chamber 13 according to the first embodiment.
Further, as a flow path penetrating in the plate thickness direction of the chip body 22, the vertical hole 2b according to the first embodiment is removed, and a sample mixing chamber 34 is added.

The first reaction chamber 30 is similar to the first reaction chamber 10 according to the first embodiment in that the elongated grooves meander in the annular region with respect to the circumferential direction of the annular region. However, the arrangement of the annular regions is different from the meandering channel structure.
The annular region in which the first reaction chamber 30 is disposed is a region sandwiched between a circle C11 that is a virtual concentric circle centering on the injection hole 2a and a circle C12 having a larger diameter than the circle C11. The circle C11 has substantially the same diameter as the circle C1 according to the first embodiment, but the circle C12 has a larger diameter than the circle C2 according to the first embodiment.
In the following, it is assumed that the circle C11 is an inscribed circle of the region where the first reaction chamber 30 is formed, and the circle C12 is a circumscribed circle of the region where the first reaction chamber 30 is formed.

As the meandering channel structure, a V-shaped bent channel 30A having a bent portion in the vicinity of the circle C12 and gradually increasing in width radially inward is disposed adjacent to the circumferential direction. Further, the meandering flow path structure has a repetitive flow path structure in which the end portions in the vicinity of the circle C11 of each bent flow path 30A are connected by an arcuate inner peripheral bent flow path 30B.
The bent channel 30A has a line-symmetric shape in which the central axis of the bent channel 30A is aligned on the diameters of the circles C11 and C12. For this reason, the two linear flow paths of each bent flow path 30A are inclined in different directions with respect to the central axis O30.

The radially inner end of the starting end 30a of the first reaction chamber 30 communicates with the introduction path 2h extending to the circle C11.
The radially inner end of the end portion 30b of the first reaction chamber 30 communicates with the end portion 34a of the sample mixing chamber 34 disposed between the start end portion 30a and the end portion 30b in the vicinity of the circle C11. Yes.
As shown in FIG. 5, the groove depth of the first reaction chamber 30 is h10 similar to that of the first embodiment.
In addition, the groove width of the first reaction chamber 30 may be different depending on the part, but in the present embodiment, as an example, all of them have the same groove width.

A plurality of second reaction chambers 32 (second reaction chamber groups) are provided along the circumferential direction of the circle C12 on the second surface 2g outside the region where the first reaction chamber 30 is provided (outside the circle C12). The pieces are provided at equal pitches (equal intervals).
Each of the second reaction chambers 32 has a common structure, and is a groove having a circular shape in plan view.

A sample solution (first reaction solution) is transferred from the measuring chamber 33 to the second reaction chamber 32 at the radially inner end of the second reaction chamber 32, so that it is shallower and narrower than the second reaction chamber 32. A transfer path 22k, which is a groove, is connected.
In this embodiment, the transfer path 22k is extended along the radial direction of the circle C12 from the vertical hole 2e formed between the second reaction chamber 32 and the circle C12.
The groove depths of the second reaction chamber 32 and the transfer path 22k can be appropriately set. In this embodiment, as an example, the depth of the second reaction chamber 12 is the same as in the first embodiment. Is set to h12, and the depth of the transfer path 2k is set to h2k.

As shown in FIG. 15, the measuring chamber 33 includes a measuring channel 33b and a branch channel 33F instead of the measuring channel 13b and the branch channel 13F of the measuring chamber 13 according to the first embodiment.
The groove depth of the measuring chamber 33 is constant at the depth h13 in the same manner as the measuring chamber 13 according to the first embodiment.

The measurement channel 33b is similar to the measurement channel 13b according to the first embodiment in that the elongated groove portion meanders in the annular region with respect to the circumferential direction of the annular region. The arrangement of the annular region is different from the meandering channel structure.
The annular region in which the measurement flow path 33b is arranged is a region sandwiched between a circle C13 that is a virtual concentric circle centering on the injection hole 2a and a circle C14 having a larger diameter than the circle C13. The circles C13 and C14 have the same diameter as the circles C11 and C12, respectively.
In the following, it is assumed that the circle C13 is an inscribed circle of the area where the measuring flow path 33b is formed, and the circle C14 is a circumscribed circle of the area where the measuring flow path 33b is formed.

The start end portion 33a of the measuring channel 33b communicates with the end portion 34b of the sample mixing chamber 34 in the vicinity of the circle C11.
As shown in FIG. 16, the end portion 33c of the measurement flow path 33b communicates with the air passage 35 extending from the first surface 2f to a position directly below the air hole 3b of the upper lid 3.
For this reason, as shown in FIG. 15, the metering flow path 33b flows substantially once in the clockwise direction from the start end portion 33a to the end portion 33c in an annular region sandwiched between the circles C13 and C14. Forming a road.
With such an arrangement, the metering channel 33b is located in a region where a part of the metering channel 33b overlaps the first reaction chamber 30 in the plate thickness direction of the chip body 22.

The metering flow path 33b is directed from the start end portion 33a toward the end end portion 33c (clockwise in FIG. 15), the first straight flow path 33A, the outer peripheral side bent flow path 33B, the second straight flow path 33C, and the inner peripheral side bent. The flow path 33D has a meandering shape that repeats in this order.
In addition, the start end part 33a of the measurement flow path 33b is the first straight flow path 33A, and the terminal end part 33c is the inner circumferential side bent flow path 33D.

The first straight flow path 33A, the outer peripheral bent flow path 33B, and the second straight flow path 33C constitute a U-shaped flow path as a whole, and the U-shaped opening is directed to the circle C13. An outer peripheral bent flow path 33B, which is a letter-shaped bent portion, is arranged inscribed in the circle C14.
Since the sample liquid (first reaction liquid) stored in the first straight flow path 33A, the outer peripheral bent flow path 33B, and the second straight flow path 33C is collectively moved to one second reaction chamber 32, The volumes of the first straight flow path 33A, the outer peripheral side bent flow path 33B, and the second straight flow path 33C are appropriately set according to the measurement amount of the sample liquid.
The first straight flow path 33A and the second straight flow path 33C are axisymmetric with respect to a central axis O33 passing through the bending center of the outer peripheral bending flow path 33B and the center of the injection hole 2a. Further, the angle formed between the first straight flow path 33A and the central axis O33 and the angle formed between the second straight flow path 33C and the central axis O33 are φ. The angle φ is preferably 0 ° or more and 60 ° or less.

The groove widths of the first straight channel 33A and the second straight channel 33C increase in diameter from the radially inner side toward the radially outer side.
The outer peripheral bent flow path 33B is an arc-shaped flow path that smoothly connects the first straight flow path 33A and the second straight flow path 33C.
The outer peripheral bending channel 13B has the largest groove width at the outermost peripheral portion in the radial direction.

The inner circumferential bent flow path 33D is a second straight line that is an adjacent flow path among the U-shaped first straight flow path 33A, the outer peripheral bent flow path 33B, and the second straight flow path 33C. It is a groove part which makes the edge part of the radial inside of the flow path 33C and the 1st linear flow path 33A communicate in circular arc shape with the same groove width.

The branch path 33F is a long and narrow groove extending radially outward along the radial direction from the portion of the outer peripheral bent flow path 33B constituting the outermost peripheral portion of the measurement flow path 33b.
The groove width of the branch path 33F is preferably the same as that of the transfer path 22k, for example.
As shown in FIG. 11, the end of each branch path 33F in the extending direction communicates with the vertical hole 2e to which the transfer path 22k is connected.
The branch path 33F and the transfer path 22k communicated via the vertical hole 2e may have the same or different extending directions. In the present embodiment, as an example, an example in which the branch path 33F and the transfer path 22k have the same extending direction along the radial direction is shown. For this reason, the branch path 33F and the transfer path 22k are aligned in the same straight line along the radial direction in plan view.

As shown in FIGS. 14 and 15, the sample mixing chamber 34 is a hole having a U shape in plan view and penetrating in the thickness direction of the chip body 22. The U-shaped opening of the sample mixing chamber 34 opens toward the circle C11 (C13), and the U-shaped bent portion is positioned in the vicinity of the circle C12 (C14).
As shown in FIG. 17, the end portion 30b of the first reaction chamber 30 is connected to the end portion 34a of the sample mixing chamber 34 at the second surface 2g. The end 34b of the sample mixing chamber 34 is connected to the start end 33a of the measuring chamber 33 on the first surface 2f.
The sample mixing chamber 34 is provided as a buffer space between the first reaction chamber 30 and the measuring chamber 33. For this reason, the volume of the sample mixing chamber 34 is at least large enough to store all the sample liquid stored in the first reaction chamber 30.
By providing such a sample mixing chamber 34, the sample liquid (first reaction liquid) is supplied before the sample liquid (first reaction liquid) sent from the first reaction chamber 30 is sent to the measuring chamber 33. When introduced into the sample mixing chamber 34, the sample liquid (first reaction liquid) sequentially fed in the space in the sample mixing chamber 34 that is much wider than the end portion 30b is mixed.
Thereby, even if unevenness (variation) occurs in the first reaction (first reaction liquid) in the first reaction chamber 30, the sample liquid (first reaction liquid) is stored in the sample mixing chamber 34 to store the sample. Since the mixing of the liquid (first reaction liquid) is promoted, the sample liquid (first reaction liquid) can be made uniform.

With such a configuration, a plurality of flow channel structures that are substantially the same as the sample analysis chip 1 according to the first embodiment are formed inside the sample analysis chip 21.
For example, when a fluid is injected from the injection port 3a, the fluid descends to the second surface 2g side through the injection hole 2a, enters the start end 30a of the first reaction chamber 30 via the introduction path 2h, The intermediate part of one reaction chamber 30 is reached and reaches the terminal part 30b.
The fluid reaching the end portion 30 b is introduced into the sample mixing chamber 34. When the amount of fluid exceeds the sample mixing chamber 34, the fluid overflows from the sample mixing chamber 34 and reaches the start end portion 33 a of the measuring chamber 33.

In the measuring chamber 33, a branch path 33 </ b> F is formed on the side of the measuring flow path 33 b, and a vent path 35 is in communication with the end portion 33 c of the measuring chamber 33.
The flow path following each branch path 33F is a flow path that goes down to the second surface 2g side through the vertical hole 2e and goes to the second reaction chamber 32 through the transfer path 22k.
Since the second reaction chamber 32 is not connected to a flow path communicating with the outside, air is confined in an unused state. For this reason, in order to move the fluid from the branch path 33F to the transfer path 22k, at least a corresponding external force that resists the air pressure must be applied to the fluid.

Since the air passage 35 is opened to the outside through the air holes 3b, the fluid in the metering flow path 33b is markedly more likely to flow through the air passage 35 than to the second reaction chamber 32 through the branch path 33F. Easy. The fluid reaching the ventilation path 35 is discharged to the outside through the ventilation path 35 and the ventilation hole 3b.

Such a sample analysis chip 21 is manufactured by manufacturing the chip body 22, the upper lid 3, and the lower lid 24 in the same manner as in the first embodiment, and appropriately fixing and attaching reagents necessary for reaction and analysis. It can be manufactured by combining them.

According to such a sample analysis chip 21, all of the sample liquid (first reaction liquid) measured in the measurement chamber 33 is moved to the second reaction chamber 32 communicated with the branch path 33F and the transfer path 22k. Except for this, sample analysis using a sample solution can be performed in the same manner as the sample analysis chip 1 according to the first embodiment.
That is, according to the sample analysis chip 21, the first and second surfaces of the plate-like portion are divided into the first reaction chamber, the measurement chamber, and the second reaction chamber (second reaction chamber group). It is easy to send and measure the sample liquid to be analyzed, and a plurality of types of biochemical reactions can be performed even with a compact configuration.

[Third Embodiment]
A sample analysis chip according to the third embodiment of the present invention will be described.
FIG. 18 is a schematic front view showing the configuration of the sample analysis chip according to the third embodiment of the present invention. FIG. 19 is a plan view of the sample analysis chip in FIG. 18 viewed from the F direction. FIG. 20 is a schematic exploded perspective view of a sample analysis chip according to the third embodiment of the present invention. FIG. 21A is a schematic plan view of a substrate used for a sample analysis chip according to a third embodiment of the present invention. FIG. 21B is a schematic back view of the base material used in the sample analysis chip according to the third embodiment of the present invention. 22 is a cross-sectional view taken along the line GG shown in FIG. 21A.

As shown in FIGS. 18, 19, and 20, the sample analysis chip 41 according to this embodiment is different from the sample analysis chip 1 according to the first embodiment that has a disk-shaped outer shape. The difference is that it has a three-dimensional outer shape.
Regarding the flow channel structure, the sample analysis chip 41 according to the present embodiment has a shape in plan view with respect to a similar flow channel structure in that a part of the flow channel structure is changed according to a change in the outer shape. However, the shape in the first embodiment is different from the mirror image.
Hereinafter, the difference between the third embodiment of the present invention and the first embodiment will be mainly described. In particular, a flow path structure that differs only in the mirror image relationship with the flow path structure according to the first embodiment is represented by adding “′” to the reference numeral according to the first embodiment, and has a detailed shape. The description of is omitted. Further, the flow path structure having the same shape as that of the first embodiment communicating with the flow path structure having such a mirror image relationship is denoted by the same reference numeral as that of the first embodiment, and the description thereof is omitted. To do.

The sample analysis chip 41 is replaced with the chip body 42 (base material), the upper lid 43 (lid material), and the lower lid instead of the chip body 2, the upper lid 3, and the lower lid 4 of the sample analysis chip 1 according to the first embodiment. 44 (cover material).

As shown in FIGS. 21A and 21B, the chip body 42 has a circular outer shape similar to the chip body 2 according to the first embodiment, and the first surface 2f and the second surface 2g are formed on the surface in the plate thickness direction. And a flat plate portion 52 (plate-like portion).
In the first surface 2f of the flat plate portion 52, a cylindrical side wall portion 51 protruding from the first surface 2f is formed on the outer peripheral portion, and has a truncated cone shape protruding from the first surface 2f in the central portion. A dispensing instrument holding base portion 54 that is a convex portion is formed.
The central axis O54 of the truncated cone-shaped outer shape of the dispensing instrument holding base 54 is coaxial with the central axis of the inner peripheral surface 51a of the side wall 51.
Further, in the first surface 2f of the flat plate portion 52, an annular region centering on the central axis O54 is provided between the inner peripheral surface 51a and the dispensing instrument holding base portion 54 in the first embodiment. A flow path structure similar to that of the first embodiment communicating with the measurement chamber 13 ′ and the measurement chamber 13 ′ in a mirror image relationship with the measurement chamber 13 is formed.

As shown in FIG. 22, a flange portion 53 that is a flange extending in parallel with the flat plate portion 52 is formed on the radially outer side at the distal end portion in the protruding direction of the side wall portion 51.
The collar portion 53 can be used for holding, positioning, position detection, etc. of the chip body 42, and can be formed with appropriate notches, through holes, concave portions, convex portions, colored surfaces, rough surfaces, etc. It is.
In the present embodiment, as shown in FIGS. 21A and 21B, as an example, an arc-shaped cutout portion 53a that is cut out in a semicircular shape from the outer periphery is formed at a position that divides the circumferential direction of the collar portion 53 into three equal parts. ing. Further, between the two arc-shaped cutout portions 53a, rectangular cutout portions 53b that are cut out in a rectangular shape from the outer periphery in order to detect the circumferential position of the chip body 42 are formed.
In addition, ribs 53 c are provided between the flange portion 53 and the outer peripheral surface of the side wall portion 51 at three locations that divide the circumferential direction into three equal parts.

As shown in FIG. 22, the dispensing instrument holding base portion 54 has a tapered surface 54 a on the outer peripheral portion, and a flat upper surface portion 54 b that is parallel to the flat plate portion 52 is formed at the tip end portion in the protruding direction.
An injection hole 54A (sample injection port) for injecting a sample liquid is provided in the central part of the dispensing instrument holding base 54 so as to penetrate in the same manner as the injection hole 2a according to the first embodiment. Yes.
In the injection hole 54A, a first taper portion 54d (taper hole portion) and a second taper portion 54e (taper hole portion) are formed in this order from the upper surface portion 54b toward the second surface 2g.

The first tapered portion 54d is a hole having a tapered shape that opens to the surface of the upper surface portion 54b and decreases in diameter toward the second surface 2g side.
The second taper portion 54e is connected to the end portion on the second surface 2g side of the first taper portion 54d, and has a tapered shape whose diameter decreases with a taper angle smaller than that of the first taper portion 54d toward the second surface 2g. It is a hole which has.

The detailed shape of the dispensing instrument holding base 54 such as the height from the upper surface 54b to the first surface 2f, the ratio of the length of the first taper 54d and the length of the second taper 54e, and the respective taper angles are as follows. , And can be set as necessary.
For example, an appropriate shape that can easily hold the tip of a dispensing device (not shown) that is used for injecting a sample liquid or other fluid into the sample analysis chip 41 can be used.
Moreover, the insertion amount of the dispensing device is also set as necessary. In the injection hole 54A, the side of the transfer path 2k from the insertion position of the dispensing device (portion close to the transfer path 2k) constitutes a flow path of the fluid to be injected.
In this embodiment, the height from the upper surface part 54b to the first surface 2f of the dispensing instrument holding base part 54 is higher than the height of the collar part 53 so that the length of the injection hole 54A can be sufficiently secured. I am trying.
The second tapered portion 54e is preferably matched to the outer shape of the distal end portion of the dispensing device so that at least a part on the first tapered portion 54d side can be held in close contact with the side surface of the dispensing device.
The first taper portion 54d has a shape that opens sufficiently larger than the outer shape of the distal end portion of the dispensing instrument so that the dispensing instrument can be easily inserted, or a shape in which the side surface of the dispensing instrument can be locked or closely attached. Is possible.
When the shape of either the second taper portion 54e or the first taper portion 54d is a shape that is in close contact with the side surface of the dispensing device, leakage of fluid injected by the dispensing device to the outside can be prevented.

In the present embodiment, the tip end portion (the lower end portion of FIG. 22) of the second taper portion 54e is formed coaxially with the central axis O54, as shown in FIG. 21B.
Further, the second tapered portion 54e communicates with the introduction path 2h similar to that of the first embodiment formed along the radial direction from the central axis O54 on the second surface 2g of the chip body 42.

As shown in FIG. 21B, a surplus liquid storage chamber 54f formed as a recess with respect to the second surface 2g by stealing meat is provided at the center of the second surface 2g.
The surplus liquid storage chamber 54f, except that a wall-like portion 54g extending from the back surface of the upper surface portion 54b to a position close to the second surface 2g is formed in the range surrounding the injection hole 54A and on the back surface of the introduction path 2h. For example, it is a recess having a shape hollowed out from the second surface 2g into a substantially truncated cone shape.
As shown in FIG. 22, in the vicinity of the injection hole 54A, a vent hole 54c, which is a hole penetrating the upper surface portion 54b and communicating with the surplus liquid storage chamber 54f, is formed.

On the outermost peripheral portion of the second surface 2 g of the chip body 42, a ridge portion 55 that protrudes from the second surface 2 g and extends along the outer peripheral surface of the side wall portion 51 is formed.
The protruding portion 55 can have an appropriate shape that can position an outer peripheral portion of a lower lid 44 described later. For example, the planar view shape may be circular, or a plurality of arc-shaped protrusions may be provided. In the present embodiment, as an example, a pair of substantially semicircular ridge portions 55 are provided so as to be spaced apart from each other at two locations in the radial direction.

Between the surplus liquid storage chamber 54f and the protrusion 55, in the annular region centered on the central axis O54, the first reaction chamber 10 and the second reaction chamber 12 (the second reaction chamber 12 according to the first embodiment). 1st reaction chamber 10 'and 2nd reaction chamber 12' (3rd implementation) which has the relationship of a mirror image with the 2nd reaction chamber group in 1 embodiment), the excess liquid storage chamber 14, the transfer path 2k, and the branch path 2m. Second reaction chamber group), surplus liquid storage chamber 14 ', transfer path 2k', branch path 2m ', and these first reaction chamber 10', second reaction chamber 12 ', surplus liquid storage chamber 14', transfer A flow path structure similar to that of the first embodiment communicating with the path 2k ′ and the branch path 2m ′ is formed.
Further, an air passage 11 ′ having a mirror image relationship with the air passage 11 according to the first embodiment is formed on the side of the introduction passage 2h. In the air passage 11 ′, the end of the second surface 2 g near the outer periphery (at a position close to the outer periphery) communicates with the vertical hole 2 d that penetrates the flat plate portion 52 and communicates with the terminal end portion 13 c ′.
Unlike the first embodiment, the end of the ventilation path 11 ′ on the second surface 2g near the inner periphery (position close to the inner periphery) communicates with the excess liquid storage chamber 54f.

The chip body 42 having such a configuration can be manufactured by using the same material as that of the chip body 2 according to the first embodiment and by the same manufacturing method as the chip body 2.

As shown in FIG. 20, the upper lid 43 is a lid member having an annular shape for sealing the flow path structure formed on the first surface 2 f of the chip body 42, according to the first embodiment. It is comprised with the material similar to the upper cover 3. FIG.
The outer diameter of the outer peripheral portion 43 b of the upper lid 43 is smaller than the inner peripheral surface 51 a of the side wall portion 51. The hole 43 a penetrating through the center of the upper lid 43 is a circular hole having a diameter larger than that of the dispensing instrument holding base 54.
The upper lid 43 is attached to the first surface 2f in the same manner as the upper lid 3 according to the first embodiment.
In this way, the upper lid 43 is formed in an annular shape so that the dispensing instrument holding base 54 has a structure that penetrates the hole 43a, so that the upper lid 43 has the same structure as in the first embodiment. Since the injection hole 2a and the vertical hole 2b can be omitted, they can be easily attached to the first surface 2f without matching the positions of the holes.

The lower lid 44 is a disk-shaped lid member that is attached to the second surface 2g of the chip body 42 and seals the flow path structure formed on the second surface 2g. It is comprised with the material similar to the lower cover 4 which concerns on a form.
The outer diameter of the outer peripheral portion 44 b of the lower lid 44 has an inner diameter that can be inserted inside the protrusion 55.
The lower lid 44 is attached to the second surface 2g in the same manner as the lower lid 4 according to the first embodiment.

According to the sample analysis chip 41 having such a configuration, an appropriate dispensing instrument can be inserted into the injection hole 54A, and a fluid such as a sample solution can be introduced into the flow path structure of the sample analysis chip 41.
For this reason, by forming the shape of the first taper portion 54d and the second taper portion 54e into a shape that can be stably held by the dispensing device, it becomes easy to dispense fluid such as sample liquid, Air leakage can be suppressed.
In particular, in the present embodiment, the dispensing instrument holding base 54 has a truncated cone shape supported by the tapered surface 54a that is inclined radially outward from the outer peripheral portion of the upper surface portion 54b. A support structure is obtained.

Next, the flow path structure formed on the sample analysis chip 41 will be described focusing on differences from the first embodiment.
For example, when a fluid is injected from a dispensing tool (not shown) inserted into the injection hole 54A, the fluid enters the start end portion 10a ′ of the first reaction chamber 10 ′ via the introduction path 2h, and enters the first reaction chamber. The terminal part 10b ′ is reached through the middle part 10 ′. At that time, since the first reaction chamber 10 ′ is in a mirror image relationship with the first reaction chamber 10 according to the first embodiment, only the point that the fluid moving direction is a mirror image is different from that in the first embodiment. Different.
The fluid that has reached the end portion 10b ′ moves up the vertical hole 2c and reaches the start end portion 13a ′ of the measuring chamber 13 ′.
Inside the measuring chamber 13 ′, the measuring chamber 13 ′ has a mirror image relationship with the measuring chamber 13 according to the first embodiment, and therefore only the point in which the fluid moving direction is a mirror image relationship is the first embodiment. Different from form.

The air passage 11 ′ communicates with the terminal portion 13c ′ of the measuring chamber 13 ′ through the vertical hole 2d, and also communicates with the surplus liquid storage chamber 54f that communicates with the outside through the air hole 54c.
For this reason, the fluid that has reached the end portion 13c ′ is discharged to the surplus liquid storage chamber 54f through the air passage 11 ′ when further pressure is applied.
The air discharged into the surplus liquid storage chamber 54f is discharged above the dispensing instrument holding base 54 through the vent hole 54c.
Moreover, it is also possible to discharge | emit liquids, such as an excess sample liquid (1st reaction liquid), to the excess liquid storage chamber 54f. In this case, the liquid such as the sample liquid (first reaction liquid) discharged into the surplus liquid storage chamber 54f is stored in the surplus liquid storage chamber 54f with the volume of the surplus liquid storage chamber 54f as a limit.
For this reason, it is possible to store all the excess liquid in advance by setting the volume of the excess liquid storage chamber 54f to be equal to or larger than the volume of the liquid to be discharged.
Thus, since the liquid discharged from the measurement chamber 13 ′ is stored from below in the surplus liquid storage chamber 54f, the clogging of the vent hole 54c is prevented and the liquid is difficult to leak out. Yes.

Further, by making the surplus liquid storage chamber 54f a sufficiently large space with respect to the cross section of the air flow path 11 ′, the vertical relationship between the lid solution and the sample liquid (first reaction liquid) is changed due to the difference in specific gravity. The lid solution covers the surface of the sample liquid (first reaction liquid) in the surplus liquid storage chamber 54f. For this reason, it is possible to improve the lid performance (covering performance) of the sample liquid (first reaction liquid) with the lid solution or the like, and the phenomenon that the sample liquid (first reaction liquid) overflows from the vent hole 54c can be suppressed. . The volume at which the lid solution and sample solution (first reaction solution) are exchanged varies depending on the solution, but the sample solution (first reaction solution) has a specific gravity of approximately water, and the lid solution has a specific gravity of 0.8 to 0, such as oil. In the case of the solution of .9, replacement occurs even in a space having a width of about 10 μL to 500 μL.

The surplus liquid storage chamber 54f is formed as a recess on the back surface of the dispensing instrument holding base 54 protruding from the first surface 2f, and is located in the central region inside the first reaction chamber 10 'and the measurement chamber 13'. Formed in space.
For this reason, by setting the protruding amount and outer diameter of the dispensing instrument holding base 54 to an appropriate size, the volume of the excess liquid storage chamber 54f can be variously changed without expanding the outer diameter of the sample analysis chip 41. Can be formed in size.
Thereby, the outer diameter of the sample analysis chip 41 is provided even when the sample liquid and the chemical liquid used for the analysis are different and the discharge amount is different by providing the mounting shape to the sample analysis device on the outer periphery of the sample analysis chip 41. Can be easily shared.

As described above, in the sample analysis chip 41, the injection hole 54A that is the sample injection port and the surplus liquid storage chamber 54f include the first reaction chamber 10 ', the measurement chamber 13', and the second reaction chamber 12 '(second It is provided in the central area surrounded by the reaction chamber group.
For this reason, as in the first embodiment, the central axis of the centrifugal axis is more than the flow path of the first reaction chamber 10 ′, the measurement chamber 13 ′, and the second reaction chamber 12 ′ (second reaction chamber group). An injection hole 54A and a vent hole 54c, which are openings to the outside air, are present at close positions. For this reason, when the sample analysis chip 41 is centrifugally rotated, back flow from the oil or sample liquid flow path to the injection hole 54A and the vent hole 54c is prevented, and excess air and bubbles are easily removed from the substrate. Can do.

Such a sample analysis chip 41 is manufactured by manufacturing the chip body 42, the upper lid 43, and the lower lid 44 in the same manner as in the first embodiment, and appropriately fixing and attaching reagents necessary for reaction and analysis. It can be manufactured by combining them.

In such a sample analysis chip 41, the flow path structure formed in the flat plate part 52 differs in the central region of the flat plate part 52, and the fluid in the flow path structure such as the first reaction chamber 10 ′ and the measurement chamber 13 ′. A sample using a sample solution in the same manner as the sample analysis chip 1 according to the first embodiment, except that the moving direction is in a mirror image relationship with the flow channel structure according to the first embodiment. Analysis can be performed.
That is, according to the sample analysis chip 41, the first surface and the second surface of the plate-like portion are separated (separately), the first reaction chamber, the measurement chamber, and the second reaction chamber (second reaction chamber group). Therefore, it is easy to send and measure the sample liquid to be analyzed, and a plurality of types of biochemical reactions can be performed even with a compact configuration.

In the description of each of the above embodiments, the measurement chamber is formed on the first surface of the plate-like portion, and the first reaction chamber and the second reaction chamber (second reaction chamber group) are formed on the second surface. As described in the example, any one of the first reaction chamber, the measurement chamber, and the second reaction chamber (second reaction chamber group) is formed on the first surface, and the other is formed on the second surface. (The first reaction chamber, the measurement chamber, and the second reaction chamber (second reaction chamber group) that are not formed on the first surface) may be formed.
For example, it is possible to form a first reaction chamber on the first surface and a measuring chamber and a second reaction chamber (second reaction chamber group) on the second surface. In addition, it is possible to form a second reaction chamber (second reaction chamber group) on the first surface and a first reaction chamber and a measuring chamber on the second surface.

In each of the above-described embodiments, the sample analysis is performed in a state where the plate-like portion of the sample analysis chip is horizontally arranged, and the example in which the first surface is upward and the second surface is downward is described. However, the sample analysis chip may be arranged such that the plate-like portion is inclined with respect to the horizontal plane. Further, the orientations of the first surface and the second surface are not limited to the orientations of the above embodiments.

In the description according to each of the above-described embodiments, the flow path structure provided on the first surface and the flow path structure provided on the second surface are described as examples in the case where the vertical holes communicate with each other. However, the channel that connects the channel structure provided on the first surface and the channel structure provided on the second surface is not limited to a vertical hole. For example, the flow path structure provided on the first surface and the flow path structure provided on the second surface may be communicated by a hole inclined with respect to the plate thickness direction of the plate-like portion, You may communicate by the flow path bent with respect to the plate | board thickness direction of a shape part.

In the description according to each of the above embodiments, the planar shape of the plate-like portion is described as an example in the case of a circle or an annular shape (donut type), but it is essential that the shape of the plate-like portion is a circle or an annular shape. is not.
For example, an appropriate shape such as a rectangular shape, an elliptical shape, a polygonal shape, or an indefinite shape can be adopted.
Further, regarding the lid member, it is not essential that the shape in plan view is a circular shape or an annular shape.
For example, a shape that is the same as or similar to the shape of the plate-like portion is possible.
In addition, the lid member may have a shape unrelated to the shape of the plate-shaped portion as long as it can cover the flow path structure formed by the groove portion or the hole portion formed on the first surface or the second surface. Is possible.

In the description according to each of the embodiments described above, the base material is described as an example in the case where the base material is composed of a single member. However, the base material may be formed by joining or bonding a plurality of members. Is possible. Further, a material may be added to the base substrate to form the shape of the substrate.

In the description according to the first and third embodiments, when the sample analysis chip has an excess liquid storage chamber for storing the fluid overflowing from the second reaction chamber, the excess liquid storage for storing the fluid overflowing from the measurement chamber. An example in the case of having a chamber has been described.
However, the excess liquid storage chamber may store excess fluid overflowing from the first reaction chamber.

In the description according to each of the above embodiments, as an example, the case where the base material is composed of the same material has been described. However, the base material can be configured by combining a plurality of types of materials.
For example, if the base material is composed of a composite material of a resin material and a metal material, the thermal conductivity of the part made of the metal material is different from the part made of the resin material, so that the temperature characteristics are changed for each part of the base material. Is possible. For example, it is possible to use a metal material for a portion where heat dissipation or heating is performed, and a resin material for a portion where it is preferable to reduce temperature change or to insulate.
The site | parts from which a material differs may be provided in a plate-shaped part, and a material may be changed with a site | part other than a plate-shaped part and a plate-shaped part.
For example, in the chip body 42 according to the third embodiment, the material of one or more portions of the side wall portion 51, the collar portion 53, and the dispensing instrument holding base portion 54 and the portion of the flat plate portion 52 is changed. Configuration is possible.
The material can also be selected based on differences in strength and rigidity.

All the components described in the above embodiments can be implemented by appropriately changing or removing combinations within the scope of the technical idea of the present invention.
For example, in all sample analysis exemplified in the first embodiment, the sample analysis chip according to the second and third embodiments can be used.
For example, in the first and third embodiments, as in the second embodiment, a configuration in which the excess liquid storage chamber 14 and the branch path 2m are removed is possible.

The sample analysis chip of the present invention can be used for detection and analysis of biochemical substances in samples such as nucleic acids. In particular, it can be widely used from detection of mutations in SNPs to techniques for detecting mutations in genes such as cancer, germ cells and somatic cells. It can also be used as a container for mixing a plurality of solutions or as a reaction container.

1, 21, 41

Sample analysis chip

2, 22 Chip body (plate-like part, base material)

2a Injection hole

2b, 2c, 2d, 2e Vertical hole 2f First surface

2g Second surface

2k, 2k ',

22k Transfer path

3, 43 Upper lid (lid material)
3a Inlet (sample inlet)
3b,

54c Ventilation holes

4, 24, 44 Lower lid (lid material)
10, 10 ', 30

First reaction chamber

10a, 10a', 30a

Start end portion

10b, 10b ',

30b End portion

11, 11', 35

Air passage

12, 12 ', 32

Second reaction chamber

13, 13', 33

Metering chambers

13F,

33F Branch channels

13a, 13a ′, 33a

Start end portions

13b,

33b Metering channels

13c, 13c ′,

33c End portions

14, 14 ′ Surplus liquid storage chamber 34 Sample mixing chamber 42 Chip body (base material)
52 Flat part (plate part)
54 Dispensing instrument holder (convex)
54a Tapered surface 54A Injection hole (sample injection port)
54d 1st taper part (taper hole part)
54e 2nd taper part (taper hole part)
54f Surplus liquid storage chamber

Claims

A sample analysis chip,
A substrate having a plate-like portion sandwiched between a first surface and a second surface;
A first reaction chamber provided in the plate-like portion for storing the sample liquid and causing a first reaction to the whole of the sample liquid;
The plate-like portion communicates with the first reaction chamber, has a plurality of branch paths for measuring the first reaction liquid obtained by causing the first reaction to occur in the sample liquid in the first reaction chamber, A weighing chamber formed in
A plurality of second reaction chambers formed in the plate-like portion to communicate with the plurality of branch paths and to individually generate a second reaction with respect to the first reaction liquid moved through the plurality of branch paths. A second reaction chamber group having
With
One of the first reaction chamber, the weighing chamber, and the second reaction chamber group is provided on the first surface,
Of the first reaction chamber, the weighing chamber, and the second reaction chamber group, a chamber that is not provided on the first surface is provided on the second surface,
Sample analysis chip.
Of the first reaction chamber, the weighing chamber, and the second reaction chamber group, at least a part of the chamber provided on the first surface and the chamber provided on the second surface are plate-shaped. The sample analysis chip according to claim 1, wherein the sample analysis chip is arranged so as to overlap in a plate thickness direction of the portion.
The weighing chamber is provided on the first surface;
The sample analysis chip according to claim 1, wherein the first reaction chamber is provided on the second surface.
The sample analysis chip according to claim 3, wherein the plurality of second reaction chambers are provided on the second surface.
Each flow path of the plurality of branch paths provided in the measurement chamber is paired with each chamber of the plurality of second reaction chambers by a plurality of vertical holes extending in the plate thickness direction of the plate-like portion. The sample analysis chip according to claim 4, wherein the sample analysis chip is in communication with one another.
The first reaction chamber includes
The sample analysis chip according to any one of claims 1 to 5, further comprising a groove portion meandering in a circumferential direction in an annular region and having an elongated shape.
The first reaction liquid communicated with the first reaction chamber and the measurement chamber is stored before the first reaction liquid sent from the first reaction chamber is sent to the measurement chamber, and the components of the first reaction liquid are stored. The sample analysis chip according to any one of claims 1 to 6, further comprising a sample mixing chamber for promoting mixing.
A central region surrounded by the first reaction chamber, the metering chamber, and the second reaction chamber group;
A sample inlet for feeding the sample solution to the first reaction chamber in the central region;
An excess liquid storage chamber for storing excess liquid overflowing from any of the first reaction chamber, the metering chamber, and the second reaction chamber group;
The sample analysis chip according to any one of claims 1 to 7, further comprising:
A convex portion having a frustoconical outer shape whose diameter decreases along a protruding direction protruding from the plate-like portion in the central region;
In the center of the convex portion, the sample injection port having a tapered hole portion whose diameter decreases in a direction opposite to the protruding direction of the convex portion and is capable of supporting a dispensing device is provided,
The sample analysis chip according to claim 8, wherein the excess liquid storage chamber is formed by a recess formed in the convex portion in a direction opposite to the protruding direction of the convex portion.
The sample analysis chip according to claim 8 or 9, wherein the excess liquid storage chamber includes an air passage that opens to the outside.
A plurality of lid members attached to the surface of the plate-like portion so as to cover each of the first reaction chamber, the measurement chamber, and the second reaction chamber group. The sample analysis chip according to Item.
The plurality of lid members are:
The sample analysis chip according to claim 11, comprising a metal lid member provided with a metal sheet-like member.
The plurality of lid members are:
The sample analysis chip according to claim 11, comprising a composite lid member provided with a composite material of a metal and a resin film.
Including a light-absorbing material between the metal sheet and the resin film, and including the composite lid material in which the metal sheet and the resin film are bonded via the light-absorbing material.
The sample analysis chip according to claim 13.
The plate-like portion is formed of a resin material,
The sample analysis chip according to any one of claims 1 to 14.