JP2009002933A - Analytical medium - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an analytical medium for smoothly sending liquid, while preventing evaporation of a sample solution passing through a flow passage, in the analytical medium having a rotatable structure and arranging the flow passage for passing the sample solution on the inside. <P>SOLUTION: This analytical medium 10 is provided with a sample filling port 20, a supplying liquid storage part 22 communicating with this sample filling port 20, a receiving liquid storage part 23 positioned on the outer peripheral side of the medium more than this supplying liquid storage part 22, and the flow passage 17 communicating with the supplying liquid storage part 22 and the receiving liquid storage part 23, on a base board, and is constituted so that a sample flows to the receiving liquid storage part 23 via the flow passage 17 from the supplying liquid storage part 22 by its centrifugal force by rotating the analytical medium 10 in a state of sealing connection ports 15 and 16 for communicating the flow passage 17 with an external part by sealing films 18 and 19. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to an analysis medium having a rotatable structure and provided with a flow path through which a sample solution can pass.

  Conventionally, each step of analyzing a sample requires an operation with an independent apparatus, which is troublesome. Each process is often a batch process, and the working time increases in proportion to the increase in the number of samples.

  In recent years, as a technique for reducing the work time by reducing the labor of each process of sample analysis, a micro total analysis system (hereinafter referred to as “μTAS”) or a lab chip (Lab-on-a) -chip: a device called "LOC") has appeared. These techniques attempt to reproduce a series of processes related to analysis such as each operation and sample movement between operations, which have been performed manually, on a single substrate. Specifically, a narrow flow path is formed on the substrate, liquid flow is controlled by branching or merging of the flow path, valves, etc., reaction control by mixing, heating, cooling, etc., detection using spectroscopic or electrical action Are devices formed on one substrate.

  One of the problems with μTAS devices is the problem of driving force for feeding a sample solution. That is, it is desired to send liquid without using a pump for liquid feeding which has a limit in miniaturization.

  Regarding such a problem, the following Patent Document 1 discloses that a polymerase chain reaction (in which a reaction solution is sent so as to alternately pass through regions having different temperatures in a flow path formed in a rotating analysis disk). An analytical disk for proceeding Polymerase chain reaction (hereinafter abbreviated as “PCR”) is disclosed. Further, Patent Document 2 below discloses a micro system platform for performing micro analysis suitable for PCR.

In these prior arts, a configuration is employed in which a solution is injected into a fine flow path formed in the disk or platform, and the sample solution injected into the flow path is sent with a centrifugal force generated by rotation. . In addition, the sample solution injected into the flow path is heated to a predetermined temperature in order to advance the reaction for analyzing the nucleic acid.
JP 2005-295877 A Special table 2003-502656 gazette

  In the analysis disk described in Patent Document 1, since the air hole and the injection port are provided at the end of the flow path, the air flows into the last side of the solution along with the liquid feeding, and the air is discharged from the front side. As a result, air is not compressed or expanded in the flow path, and liquid feeding is smooth. However, it was not possible to prevent the ejection of liquid or vapor in the flow path.

  In addition, in the method and apparatus for performing microanalysis described in Patent Document 2, the end of the flow path has an air channel that is thin enough to pass only air holes and air but not liquid. Therefore, it is possible to prevent ejection of liquid while allowing air to be discharged when the solution is fed. However, even with this technique, it was still impossible to prevent the ejection of steam. Accordingly, the moisture in the flow path is vaporized by heating or the like and exits from the air hole or the injection port, so that the concentration of the sample changes with time, and there is a possibility that it will eventually dry.

  Accordingly, an object of the present invention is to provide an analytical medium that can be used as a μTAS device, and that can smoothly feed liquid while preventing evaporation of a sample solution passing through a flow path. .

To achieve the above object, the first analytical medium of the present invention comprises a rotatable medium, a sample inlet, a supply reservoir that communicates with the sample inlet, a receiver reservoir, A flow path communicating with the supply liquid reservoir and the reception liquid reservoir is provided, and a sample flows from the supply liquid reservoir to the reception liquid reservoir through the flow path by centrifugal force due to rotation. In the analytical medium configured as follows:
The medium is mainly composed of a substrate, and the substrate includes the flow path, a first connection port communicating with one end of the flow path, and the other end of the flow path. A second connection port is formed,
A first sealing film for sealing the first connection port and a second sealing film for sealing the second connection port are attached to the substrate surface,
Between the first sealing film and the substrate surface, the supply liquid reservoir portion including a bag-like space communicating with the first connection port is formed, and the second sealing film and the substrate surface In between, the receiving liquid reservoir portion formed of a bag-like space communicating with the second connection port is formed,
The first sealing film is formed with a sample communication port that communicates with the sample injection port, the sample injection port, and the supply liquid reservoir, and the first sealing film is supplied through the sample injection port. The sample communication part can be sealed after the sample is filled in the liquid storage part.

  According to the first analytical medium, when a sample is injected from the sample injection port of the first sealing film, the sample is filled into a supply liquid reservoir portion that is formed of a bag-like space communicating with the first connection port. After injecting the sample, sealing the sample communication part communicating with the sample inlet of the first sealing film and the supply liquid reservoir part provides communication with the supply liquid reservoir part, the receiving liquid reservoir part, and the like. The sample passage composed of the flow path is sealed or closed.

  In this state, when the analysis medium is rotated and a centrifugal force is applied to the sample, the sample flows from the supply medium reservoir for the analysis medium to the reception liquid reservoir through the channel. At this time, the supply liquid reservoir portion is deflated so that the first sealing film forming the bag-like space approaches the substrate surface, and the volume is reduced. On the other hand, the receiving liquid reservoir swells so that the second sealing film forming a bag-like space in a pressure-increasing state is separated from the substrate surface, and the volume increases. By such an action, the sample can be flowed through the flow path from the supply reservoir to the receiving reservoir.

  When the sample passes through the flow path, even if the sample is heated, for example, the sample passage is sealed or closed by the first sealing film and the second sealing film, so that the solution in the sample evaporates. Therefore, it is possible to prevent the sample concentration and the like from changing. What is necessary is just to design the sealing degree of a sample channel | path suitably according to the precision of analysis, and the objective can be achieved also by sticking a sealing film etc.

  In the first analysis medium, the receiving liquid reservoir is formed on the outer peripheral side of the medium with respect to the supply liquid reservoir, and the second connection port is the first connection It is preferable to be formed so as to be positioned on the outer peripheral side of the medium with respect to the connection port. This prevents the centrifugal force applied to the sample from reaching the equilibrium state on the sample supply side and the sample reception side as the sample moves from the supply side to the reception side, thereby preventing the flow from stopping. Can do.

  In the first analysis medium, the medium is mainly composed of a substrate formed by joining a first substrate member and a second substrate member, and the first substrate member and the second substrate are formed. It is preferable that the flow path is formed between the members. According to this, the flow path can be easily formed on the substrate by molding a sculpture-like groove on at least one substrate member and joining the other substrate members so as to be bonded together.

  In the first analysis medium, the medium is mainly composed of a substrate formed by joining a substrate member and a sealing member, and the flow path is provided between the substrate member and the sealing member. Is preferably formed. According to this, the flow path can be easily formed on the substrate by molding a sculpture-shaped groove on at least one substrate member and joining the sealing member so as to stick them together.

  In the first analysis medium, it is preferable that the first sealing film can be sealed by welding or adhering the first sealing film to the substrate in the sample communication portion. According to this, after injecting the sample from the sample injection port, the portion of the flow channel connecting the sample injection port and the supply liquid reservoir is sealed by welding or bonding to the substrate, and the sample passage can be easily and It can be surely sealed or closed.

Further, the second analysis medium of the present invention includes a rotatable medium, a sample inlet, a supply reservoir that communicates with the sample inlet, a receiving reservoir, and the supply reservoir. And a flow path communicating with the receiving liquid reservoir, and the sample flows from the supply liquid reservoir through the flow path to the receiving liquid reservoir by a centrifugal force due to rotation. In analytical media,
The medium is mainly composed of a substrate, and the substrate communicates with the supply liquid reservoir, the receiving liquid reservoir, the flow path, and the supply liquid reservoir. A first connection port and a second connection port communicating with the receiving liquid reservoir,
On the surface of the substrate, one sealing film for sealing the first connection port and the second connection port is attached,
Between the sealing film and the substrate surface, a communication gap that connects the first connection port and the second connection port is formed,
The sealing film is formed with the sample inlet, and a sample communication portion that communicates with the sample inlet and the first connection port, and the sealing film passes through the sample inlet to the supply liquid reservoir. The sample communication portion can be sealed after the sample is filled.

  According to the second analysis medium, when a sample is injected from the sample injection port of the sealing film, the sample is filled into the supply liquid reservoir through the first connection port. After injecting the sample, when the sample communication part communicating with the sample injection port and the first connection port is sealed, it is constituted by a supply liquid reservoir part, a reception liquid reservoir part, and a flow path communicating therewith. The sample passage is sealed or closed.

  In this state, when the analysis medium is rotated and a centrifugal force is applied to the sample, the sample flows from the supply medium reservoir for the analysis medium to the reception liquid reservoir through the channel. At this time, in the supply reservoir, the pressure decreases due to the outflow of the sample, and in the receiving reservoir, the pressure increases due to the inflow of the sample. However, the air or the like enclosed in the receiving reservoir is connected to the second connection. From the mouth, it is received in the communication gap. Furthermore, since it flows into the supply liquid reservoir through the first connection port, the sample can flow relatively easily.

  When passing through the flow path, for example, even if the sample is heated, the sample passage is sealed or closed by the sealing film, so that the solution in the sample does not evaporate and is released to the outside air. Etc. can be prevented from changing.

  In the second analysis medium, the receiving liquid reservoir is formed on the outer peripheral side of the medium with respect to the supply liquid reservoir, and the second connection port is the first connection port. It is preferable to be formed so as to be positioned on the outer peripheral side of the medium with respect to the connection port. This prevents the centrifugal force applied to the sample from reaching the equilibrium state on the sample supply side and the sample reception side as the sample moves from the supply side to the reception side, thereby preventing the flow from stopping. Can do.

  In the second analysis medium, the medium is mainly composed of a substrate formed by joining a first substrate member and a second substrate member, and the first substrate member and the second substrate are formed. It is preferable that the flow path is formed between the members. According to this, the flow path can be easily formed on the substrate by molding a sculpture-like groove on at least one substrate member and joining the other substrate members so as to be bonded together.

  In the first analysis medium, the medium is mainly composed of a substrate formed by joining a substrate member and a sealing member, and the flow path is provided between the substrate member and the sealing member. Is preferably formed. According to this, the flow path can be easily formed on the substrate by molding a sculpture-shaped groove on at least one substrate member and joining the sealing member so as to stick them together.

  In the second analysis medium, it is preferable that the sealing film can be sealed by welding or adhering the sealing film to the substrate in the sample communication portion. According to this, after injecting the sample from the sample injection port, the portion of the flow path that connects the sample injection port and the portion that becomes the communication gap is sealed by welding or bonding to the substrate, and the sample passage can be easily and It can be surely sealed or closed.

  In the analysis medium of the present invention, the flow path connecting the supply reservoir and the receiving reservoir is alternately folded outward and inward along the radial direction of the medium, It has a zigzag part that goes in the circumferential direction as a whole, and it is preferable that the inner circumference side and the outer circumference side of the zigzag part are arranged in different temperature regions when analyzing the sample. According to this, when the sample passes through the zigzag portion of the flow path, the high temperature region and the low temperature region are alternately passed. Therefore, for example, a specific DNA existing in the sample is detected by the PCR method. It can be suitably used for an analysis method for selectively amplifying and detecting.

  As described above, according to the analysis medium of the present invention, the closed state in which the sample passage constituted by the supply liquid reservoir, the reception liquid reservoir, and the flow path communicating with them is blocked from the outside air. Thus, the sample can be flowed from the supply reservoir to the receiving reservoir through the channel, and analysis can be performed by flowing in the channel while the sample is sealed. For this reason, even when it is necessary to heat the sample when passing through the flow path, the solution or the like in the sample is not evaporated and released to the outside air, and the sample concentration or the like can be prevented from changing.

  First, with reference to FIGS. 1-6, 1st Embodiment of the medium for analysis of this invention is described.

  As shown in FIG. 1, the analysis medium 10 is configured by joining a first substrate member 11 and a second substrate member 12 made of a round disc having substantially the same diameter. The material of the first substrate member 11 and the second substrate member 12 is not particularly limited. For example, thermoplastic resins such as acrylic, polystyrene, polycarbonate, polyethylene, polypropylene, polyethylene terephthalate, polyvinyl chloride, and polyvinylidene chloride are preferably used. Is done. When analyzing the sample by the light detection means, at least one of the first substrate member 11 and the second substrate member 12 is preferably made of a light transmissive material, and the other has a reflective layer such as a metal vapor deposition film. It may be provided.

  Support holes 13 and 14 through which the rotation shafts are inserted are formed at the centers of the first substrate member 11 and the second substrate member 12, respectively. Further, the first substrate member 11 is formed with a first connection port 15 and a second connection port 16 which are formed of through holes. The first connection port 15 is arranged on the inner peripheral side of the second connection port 16, that is, at a position close to the support hole 13.

  On the inner surface (the upper surface in FIG. 1) of the second substrate member 12, a flow path 17 made of a linear groove is formed. The flow path 17 has one end 17a extending to a position corresponding to the first connection port 15 of the first substrate member 11, and the other end extending to a position corresponding to the second connection port 16 of the first substrate member 11. It has an end portion 17b and a zigzag portion 17c that is located between the respective end portions 17a and 17b and alternately turns outward and inward along the radial direction and goes in the circumferential direction as a whole.

  As shown in FIGS. 1 and 2, a first sealing film 18 covering the first connection port 15 and a second sealing film covering the second connection port 16 are provided on the surface of the first substrate member 11 (upper surface in FIG. 1). 19 is attached. FIG. 1 shows a state before sticking, and FIG. 2 shows a sticking state. A portion A of the first sealing film 18 and the second sealing film 19 with dot-like hatching represents a portion that is or has been adhered to the first substrate member 11 by means such as adhesion or welding. Further, a small circle B formed by a dotted line of the first sealing film 18 represents a portion aligned with the first connection port 15, and a small circle C formed by a dotted line of the second sealing film 19 is aligned with the second connection port 16. Represents a part.

  Further, the inside of the great circle D formed by the dotted line of the first sealing film 18 forms a supply liquid reservoir 22 including a bag-shaped space surrounding the first connection port 15 between the first substrate member 11 and the first substrate member 11. It is part. The cross-sectional structure is shown in FIG. In addition, the inside of the great circle E formed by the dotted line of the second sealing film 19 forms a receiving liquid reservoir 23 formed of a bag-like space surrounding the second connection port 16 with the first substrate member 11. It is part. The cross-sectional structure is shown in FIG.

  A sample injection port 20 is formed in the first sealing film 18, and the sample injection port 20 is connected to the first sealing member 18 through a sample communication portion 21 formed by a thin linear portion that is not attached to the first substrate member 11. The supply liquid reservoir 22 communicates with the supply liquid reservoir 22. As a result, as shown in FIG. 3, the sample inlet 20 communicates with the supply liquid reservoir 22 via the sample communication portion 21, and the supply liquid reservoir 22 passes through the first connection port 15. The channel 17 communicates with one end 17a.

  As shown in FIG. 4, the inner side of the second sealing film 19 forms a receiving liquid reservoir 23 composed of a bag-like space surrounding the second connection port 16, and the receiving liquid reservoir 23 includes It communicates with the other end 17 b of the flow path 17 through the two connection ports 16.

  The first sealing film 18 and the second sealing film 19 may be composed of a single sealing film.

  The sealing film is preferably a low melting point material such as polypropylene or polyethylene, or a high melting point material such as nylon or polyethylene terephthalate (PET). Moreover, it is preferable to use what laminated | stacked transparent orientation oxides, such as an alumina and a silica, on the surface of nylon and polyethylene terephthalate (PET) made into the laminate film.

  Thus, one sample passage comprising the sample injection port 20, the sample communication portion 21, the supply liquid reservoir 22, the first connection port 15, the flow path 17, the second connection port 16, and the reception liquid reservoir 23 is formed. It is configured. In this passage, the supply liquid reservoir 22 is located on the inner peripheral side with respect to the reception liquid reservoir 23, and the flow path 17 having the zigzag portion 17 c is disposed between the liquid reservoirs 22 and 23. The shape is made.

  As shown in FIG. 5, the analysis medium 10 is formed with a plurality of sample passages as described above along the circumferential direction, eight in the case of this embodiment, and eight at the same time. The sample can be analyzed.

  As shown in FIG. 6, in the case of this embodiment, the zigzag portion 17c of the flow path 17 in each sample passage is a high temperature region where the inner peripheral side A is set to 90 to 99 ° C., and the outer peripheral side It is comprised so that it may become a low-temperature area | region set to 80 degreeC. Each of the above temperature regions can be formed by providing a temperature control means having a heating means composed of, for example, a heating wire, a Peltier element, a lamp heater, or the like, on a turntable on which the analysis medium 10 is installed or a portion facing the turntable. it can.

  FIG. 7 shows another aspect of the zigzag portion 17 c of the flow path 17. In this embodiment, the zigzag portion 17c of the flow path 17 in each sample passage is arranged so as to be directed from the inner peripheral side to the outer peripheral side as a whole while alternately folding back left and right in the circumferential direction along the radial direction of the analysis medium 10. The In this case, each temperature region can be formed to have different temperature settings on the right and left sides in the circumferential direction by providing the temperature control means on the turntable on which the analysis medium 10 is installed.

  According to the above configuration, since the sample passing through the flow path 17 repeatedly passes through the high temperature region A and the low temperature region B, it is preferably used for an analyzer that detects DNA amplified by PCR. Can do.

  Next, a method of using the analysis medium 10 having the above configuration will be described.

  First, a predetermined amount of a sample for analysis is injected from the sample injection port 20 using a syringe (not shown). In this case, an elastic ring that is in close contact with the periphery of the sample injection port 20 is provided at the discharge port of the syringe so that the sample is injected into the inner periphery of the first sealing film 18 through the sample injection port 20 without spilling. To.

  For example, when PCR is performed as a sample, a solution containing a reagent for analyzing specific DNA amplified by the PCR method is used. Specifically, a template DNA for amplification purposes, a heat-resistant DNA polymerase that is an enzyme that synthesizes DNA complementary to the template, characterized by a high optimum temperature, and two sites selected on the template DNA A solution containing two kinds of primer DNAs having the ability to form double strands in each of the specific sequences and nucleotides serving as substrates for DNA polymerase is used.

  In addition, when performing real-time PCR, as a sample for analysis containing nucleic acid, a dye (fluorescent dye) whose fluorescence intensity is increased by binding to double-stranded DNA, or a dye / quencher is placed nearby. Fluorescence resonance energy transfer between dyes by hybridization of nucleic acid probes designed to cause dissociation of dyes / quenchers by extension reaction, or two types of nucleic acid probes to which donor dyes and acceptor dyes are added. Nucleic acid probes designed to cause the phenomenon can be included. There is no particular limitation on the fluorescent dye or nucleic acid probe mixed with the analytical sample containing nucleic acid, and it can be selected according to the application. For example, a preferred example of the fluorescent dye is SYBR Green I (Molecular Probes). Moreover, as a nucleic acid probe, the TaqMan probe (made by Roche) which has arbitrary arrangement | sequences, Hybridization Probes (made by Roche) etc. can illustrate preferably. There are no particular limitations on the origin and base sequence of the template DNA for the purpose of quantification, and it may be collected from human blood, urine, saliva, semen, milk, etc., or derived from organisms other than humans. May be.

  The sample injected from the sample injection port 20 is filled into the supply liquid reservoir 22 through the sample communication part 21. When a predetermined amount of the sample is injected, the first sealing film 18 is thermally welded to the first substrate member 11 at the portion of the sample communication portion 21 by the heater H shown in FIGS. . In addition, joining of a sealing film and a board | substrate can also be performed by methods, such as adhesion | attachment and thermocompression bonding. As a result, the passage of the sample leading to the supply liquid reservoir 22, the flow path 17, and the receiving liquid reservoir 23 is blocked from the outside air.

  Then, the analysis medium 30 is placed on a rotary table (not shown), and the temperature control means (not shown) sets the inner peripheral side of the zigzag portion 17c of the flow path 17 to a high temperature region and lowers the outer peripheral side to a low temperature. Region (see FIG. 6).

  In this state, when the rotating medium (not shown) is rotated to rotate the analysis medium 10 at a predetermined speed, the sample filled in the supply liquid reservoir 22 is moved by the centrifugal force to the first connection port 15. From the second connection port 16 to the receiving liquid reservoir 23 through the one end portion 17a of the flow path 17, the zigzag portion 17c, and the other end portion 17b. At this time, in the supply liquid reservoir 22, the first sealing film 18 shown in FIG. 3 is deflated so as to approach the first substrate member 11 and the volume is reduced. In the receiving liquid reservoir 23, the first sealing film 18 shown in FIG. Since the two-sealing film 19 swells away from the first substrate member 11 to increase the volume, the sample can flow.

  Then, when the sample passes through the zigzag portion 17c of the flow path 17, the sample is exposed to a temperature environment according to those regions by repeatedly passing through the high temperature region A and the low temperature region B. . Thus, for example, when performing PCR, the high temperature region (a) is formed so that the sample moving in the flow path can be heated to a temperature higher than the DNA dissociation temperature, and the low temperature region (B) is used for the sample moving in the flow path. It is formed so that it can be controlled below the DNA dissociation temperature. Then, the ratio of the area or flow path length of each set temperature region is formed so that the ratio of the time for which the sample passes is 1: 2 to 1: 3 in the high temperature region: low temperature region. Furthermore, by adjusting the rotation speed, the flow rate of the liquid sent by the centrifugal force can be controlled so as to pass through one temperature cycle over 10 seconds to 1 minute. In this way, the desired temperature cycle for PCR can be imparted. At this time, the passage of the sample is sealed by the first sealing film 18 and the second sealing film 19 and is sealed or closed so as to be shut off from the outside air, so that the sample passes through the zigzag portion 17c of the flow path 17. Even if heated, the vapor does not flow out to the outside, and the concentration of the sample does not change.

  In the above embodiment, the flow path 17 is formed in a groove shape on the bonding side surface of the second substrate member 12, but the flow path 17 is formed in a groove shape on the bonding side back surface of the first substrate member 11. You may form it.

  FIG. 8 shows another aspect of the analysis medium 10. In this embodiment, the channel 17 is formed in a groove shape on the back surface (the lower surface in FIG. 8) of the substrate member 11a. Then, the entire first connection port 15, the second connection port 16, and the flow path 17 formed in the substrate member 11a are covered with the sealing member 12a from the back side, and are sealed in a state where the passage of the flow path 17 is secured. Stick like so.

  And the 1st sealing film 18 which covers the 1st connection port 15 and the 2nd sealing film 19 which covers the 2nd connection port 16 are stuck on the surface (upper surface of Drawing 8) of substrate member 11a. A portion A of the first sealing film 18 and the second sealing film 19 with dot-like hatching represents a portion that is or has been adhered to the first substrate member 11 by means such as adhesion or welding. Further, a small circle B formed by a dotted line of the first sealing film 18 represents a portion aligned with the first connection port 15, and a small circle C formed by a dotted line of the second sealing film 19 is aligned with the second connection port 16. Represents a part.

  Further, the inside of the great circle D formed by the dotted line of the first sealing film 18 is a portion that forms the supply liquid reservoir portion 22 including a bag-shaped space surrounding the first connection port 15 between the substrate member 11a. There is no. FIG. 9 shows the cross-sectional structure in a cross-sectional view in the same arrangement as the cross-sectional view taken along the line III-III in FIG. Further, the inside of the great circle E formed by the dotted line of the second sealing film 19 is a portion that forms a receiving liquid reservoir portion 23 formed of a bag-like space surrounding the second connection port 16 with the substrate member 11a. There is no. FIG. 10 shows the cross-sectional structure in a cross-sectional view in the same arrangement as the cross-sectional view taken along the line IV-IV in FIG.

  With the above configuration, the analysis medium 10 of the present invention can be configured with a single substrate member without joining the two substrate members.

  The material of the sealing member is not particularly limited, but the same material as the sealing film is preferable in order to form a state in which the passage of the sample is blocked from outside air.

  In addition, the groove for forming the flow path formed in the substrate member may be formed by printing with the same kind of synthetic resin as the substrate on a portion other than the portion corresponding to the engraved groove described above. it can. Or you may comprise the said flow path formed in the said board | substrate member by sticking the separate thing shape | molded by resin in the pipe shape to a board | substrate member.

  Next, a second embodiment of the analysis medium of the present invention will be described with reference to FIGS. Note that the same reference numerals are given to substantially the same parts as those in the embodiment described above.

  As shown in FIG. 11, the analysis medium 30 is configured by joining the first substrate member 11 and the second substrate member 12 in the same manner as in the above embodiment. Are formed with support holes 13 and 14.

  In the second substrate member 12, a supply liquid reservoir 22, a receiving liquid reservoir 23, and a flow path 17 communicating with these are formed in a groove shape. In the flow path 17, one end portion 17 a communicates with the supply liquid reservoir portion 22, the other end portion 17 b communicates with the reception liquid reservoir portion 23, and an intermediate portion forms a zigzag portion 17 c.

  Furthermore, a supply path 31 extends from the supply reservoir 22, a return path 32 extends from the receiving reservoir 23, and the end of the supply path 31 is the outer peripheral side and the end of the return path 32. Is located on the inner peripheral side, and the respective end portions are arranged close to each other.

  In the first substrate member 11, the first connection port 15 is formed at a position corresponding to the end of the supply path 31, and the second connection port 16 is formed at a position corresponding to the end of the return path 32.

  Then, referring also to FIG. 12, one sealing film 33 is attached to the surface of the first substrate member 11 so as to cover the first connection port 15 and the second connection port 16. . In the sealing film 33, a portion A with dot-shaped hatching in FIGS. 11 and 12 represents a portion attached to the first substrate member 11 by means of adhesion, welding, or the like, and also includes a dotted line. A small circle B represents a portion that matches the first connection port 15, and a small circle C that is a dotted line represents a portion that matches the second connection port 16. A sample injection port 20 is formed in the sealing film 33.

  The inside of the sealing film 33 communicates with the first connection port 15, the second connection port 16, and the sample injection port 20, and forms a supply channel for flowing a sample from the sample injection port 20 to the first connection port 15, A communication gap is formed to receive air or the like pushed out from the flow path as the sample is fed into the flow path. Therefore, the air pushed out from the second connection port 16 is received in the communication gap and further returned to the first connection port 15 communicating with the communication gap.

  That is, as shown in FIG. 13, the sample injection port 20 communicates with the first connection port 15 through the inside of the sealing film 33, and the sample injected from the sample injection port 20 is inside the sealing film 33. Through the first connection port 15 and then into the supply path 31 and into the supply liquid reservoir 22. The sample injection port 20 is also communicated with the second connection port 16, but the second connection port 16 is formed by an adhesive portion Aa extending in a partition shape between the sample injection port 20 and the second connection port 16. The sample is difficult to flow into. The position of the sample injection port 20 may be appropriately changed to a position shifted substantially parallel to the heater H shown in FIG. 11, or may be appropriately changed to a position closer to the small circle B than the position. Also good.

  Further, as shown in FIG. 14, the air or the like pushed out from the receiving liquid reservoir 23 passes through the return path 32 and flows from the second connection port 16 into the communication gap in the internal space of the sealing film 33. Further, the air enters the first connection port 15 through the communication gap in the internal space, and further returns to the supply liquid reservoir 22 through the supply path 31.

  Next, a method of using the analysis medium 30 having the above configuration will be described.

  First, a predetermined amount of a sample for analysis is injected from the sample injection port 20 using a syringe (not shown). As described above, the sample injected from the sample injection port 20 flows into the supply path 31 from the first connection port 15 through the inner space of the sealing film 33 and flows into the supply liquid reservoir 22. When the sample is injected, the sealing film 33 is thermally welded to the first substrate member 11 between the sample injection port 20 and the first connection port 15 and the second connection port 16 by, for example, the heater H, and the first connection port 15 and the second connection port 16 are sealed with a sealing film 33. At this time, a communication gap 34 is formed in the sealing film 33 to communicate the first connection port 15 and the second connection port 16 (see FIGS. 12 and 14). In addition, joining with the board | substrate of a sealing film can also be performed by methods, such as adhesion | attachment and thermocompression bonding.

  Then, the analysis medium 30 is placed on a rotary table (not shown), and the temperature control means (not shown) sets the inner peripheral side of the zigzag portion 17c of the flow path 17 to a high temperature region and lowers the outer peripheral side to a low temperature. Region (see FIG. 6).

  In this state, by rotating a rotary table (not shown) and rotating the analysis medium 30, the sample stored in the supply liquid reservoir 22 moves in the outer diameter direction by the action of centrifugal force, and the flow path 17 and flows into the receiving liquid reservoir 23. At this time, the air or the like originally filled in the receiving liquid reservoir 23 is pushed out by the sample flowing from the flow path 17, passes through the return path 32, passes through the second connection port 16, and is received in the communication gap 34. The Further, it flows into the first connection port 15, passes through the supply path 31, and is returned to the supply liquid reservoir 22. As a result, the sample can flow in the flow path 17.

  Then, when the sample passes through the zigzag portion 17c of the flow path 17, by repeatedly passing through the high temperature region A and the low temperature region B, for example, when performing PCR, A desired temperature cycle can be applied. At this time, the passage of the sample is sealed by the sealing film 33 and is sealed or closed so as to be shut off from the outside air. Therefore, even if the sample is heated when passing through the zigzag portion 17 c of the flow path 17, Thus, the sample does not flow out and the concentration of the sample does not change.

  In the above embodiment, the supply liquid reservoir 22, the flow path 17, and the reception liquid reservoir 23 are formed in a groove shape on the bonding side surface of the second substrate member 12. The first substrate member 11 may be formed in a groove shape on the bonding side surface.

  FIG. 15 shows another embodiment of the analysis medium 30 described above. In this embodiment, the channel 17 is formed in a groove shape on the back surface (the lower surface in FIG. 15) of the substrate member 11a. Then, the entire first connection port 15, the second connection port 16, and the flow path 17 formed in the substrate member 11a are covered with the sealing member 12a from the back side, and are sealed in a state where the passage of the flow path 17 is secured. Stick like so.

  A single sealing film 33 is adhered to the surface of the substrate member 11 a so as to cover the first connection port 15 and the second connection port 16. In the sealing film 33, a portion A with dot-like hatching in FIGS. 11 and 12 represents a portion attached to the substrate member 11a by means of adhesion, welding or the like, and is a small circle made of a dotted line. B represents a portion that matches the first connection port 15, and a small circle C that is a dotted line represents a portion that matches the second connection port 16. A sample injection port 20 is formed in the sealing film 33.

  The inside of the sealing film 33 communicates with the first connection port 15, the second connection port 16, and the sample injection port 20, and forms a supply channel for flowing a sample from the sample injection port 20 to the first connection port 15, A communication gap is formed to receive air or the like pushed out from the flow path as the sample is fed into the flow path. Therefore, the air pushed out from the second connection port 16 is received in the communication gap and further returned to the first connection port 15 communicating with the communication gap.

  FIG. 16 shows a cross-sectional view in the same arrangement as the cross-sectional view taken along the line III-III in FIG. As shown in FIG. 16, the sample injection port 20 communicates with the first connection port 15 through the inside of the sealing film 33 by sticking the sealing member 12a to the substrate member 11a from the back side. The sample injected from the sample injection port 20 passes through the inside of the sealing film 33, passes through the first connection port 15, flows into the supply path 31, and flows into the supply liquid reservoir 22. Yes.

  FIG. 17 shows a sectional view in the same arrangement as the sectional view taken along the line IV-IV in FIG. As shown in FIG. 17, by sticking the sealing member 12 a to the substrate member 11 a from the back side, the air or the like pushed out from the receiving liquid reservoir 23 passes through the return path 32 and passes through the second connection port 16. To the communication gap in the internal space of the sealing film 33. Further, the air enters the first connection port 15 through the communication gap in the internal space, and further returns to the supply liquid reservoir 22 through the supply path 31.

  With the above configuration, the analysis medium 30 of the present invention can be configured with a single substrate member without joining the two substrate members.

  As with the analysis medium 10 described above, the material of the ceiling member is not particularly limited. However, in order to form a state where the passage of the sample is blocked from the outside air, the same material as the sealing film is preferable. . In addition, the groove for forming the flow path formed in the substrate member may be formed by printing with the same kind of synthetic resin as the substrate on a portion other than the portion corresponding to the above-mentioned engraved groove. it can. Or you may comprise the said flow path formed in the said board | substrate member by sticking the separate thing shape | molded by resin in the pipe shape to a board | substrate member.

  As a detection means for analyzing the sample, a light detection means using an optical system similar to a general absorptiometer or fluorometer can be used. According to this, it is possible to analyze the sample without collecting the sample in the medium. For example, taking the case of performing PCR using the fluorescent dye SYBR green as an example, the fluorescent dye is inserted (intercalated) into double-stranded DNA synthesized by PCR to generate fluorescence. The amount of double-stranded DNA synthesized by PCR can be detected by measuring the fluorescence intensity using a general fluorescence excitation / detection means. In order to enable measurement at a plurality of points in the flow path, the medium may be moved to move the measurement point to a position that matches the position of the detection means, or the light detection means may be moved. Or a combination of both methods may be employed.

  Examples of the light source of the light detection means by the optical system include a xenon lamp, a halogen lamp, and a semiconductor laser. Examples of optical receivers include photomultiplier tubes, CCDs, and photodiodes. The spectroscopic means includes a prism, a filter, a grating, a slit, a beam splitter, a lens, a mirror, and the like. In measuring the absorbance, light may be passed through the measurement point or reflected. For detection of fluorescence, a light receiver may be disposed at any efficient location. Turbidity measurement is basically the same as absorbance measurement.

  Hereinafter, the present invention will be specifically described with reference to examples. The disc-shaped analysis medium manufactured in these examples is an example of the present invention, and the present invention is not limited to this.

<Production example 1>
An analytical medium having the structure shown in FIGS. 1 to 5 was manufactured under the following conditions.

  As the first substrate member 11, a polycarbonate substrate having a diameter of 3 mmφ which is the first connection port 15 and the second connection port 16 and having an outer diameter of 120 mmφ, an inner diameter of 15 mmφ and a thickness of 500 μm was molded by injection molding. In addition, the holes with a diameter of 3 mm.phi. Used as both connection ports were formed at positions matching the both end portions 17a and 17b of the flow path on the second substrate member 12 formed as described later.

  As the second substrate member 12, a polycarbonate substrate having a groove serving as a flow path 17 and having an outer diameter of 120 mmφ, an inner diameter of 15 mmφ, and a thickness of 500 μm was molded by injection molding. The shape of the flow path 17 is a flow path having a width of 200 μm and a height of 30 μm, folded back with a length of 10 mm, and having a total length of 200 mm after 10 reciprocations, and a wall between the flow paths. The width was 200 μm. And the zigzag part of the flow path 17 was arranged so that it might be located in the range of 40 mm to 50 mm from the center of a disk-shaped medium, and eight were formed along the circumferential direction.

  As the first sealing film 18, a laminate film of modified polypropylene / silica / polycarbonate (30 μm / 1 μm / 100 μm) having a hole with a diameter of 2 mmφ serving as the sample injection port 20 was punched and molded in a size of 15 mm × 15 mm. The modified polypropylene of the laminate film serves as a sealant for bonding with the substrate.

  As the second sealing film 19, a laminate film of modified polypropylene / silica / polycarbonate (30 μm / 1 μm / 100 μm) was punched and molded in a size of 15 mm × 15 mm.

  In order to obtain the arrangement shown in FIG. 1, both molded substrates were heated and pressure-bonded while being evenly pressurized. The pressure was 106 Pa, the temperature was 130 ° C., and heating and pressing were performed for 1 minute. Furthermore, from above the second substrate, the sealing films 18 and 19 are heat-sealed at 150 ° C. along the shape of the affixed portion (the portion A with dot-shaped hatching in FIGS. 1, 2 and 5). Stuck. At this time, the first sealing film 18 was adhered so that the non-adhered portion of the first sealing film 18 formed the supply liquid reservoir portion 22 of 10 mmφ and the sample communication portion 21 up to the sample injection port 20. Further, the second sealing film 19 was stuck so that the non-adhered portion of the second sealing film 19 formed a receiving liquid reservoir 23 having a diameter of 10 mm.

<Example 1>
The following tests were conducted using the analytical medium produced in Production Example 1.

  After about 10 μL of pure water is injected from the sample inlet 20 through an O-ring with a microsyringe and stored in the supply liquid reservoir 22, the sample inlet 20 and the supply liquid reservoir 22 are approximately in the middle. The sample communication part 21 was heat-sealed with a baking iron (indicated by H in the figure). In this way, pure water was injected into all eight flow paths on the analysis medium and heat sealed. As shown in FIG. 18, the analysis medium is installed in an analysis medium heating and rotating apparatus having a rotary table 42 rotated by a motor 43 and heating means including a lamp 41 arranged on the rotary table 42. The analysis medium was heated to 95 ° C. and allowed to stand for 10 minutes, and then rotated at a predetermined rotation speed and time as shown in Table 1 below. After the elapse of a predetermined time, the rotation of the analysis medium was stopped, and the position of the tip of the supplied pure water was measured as the distance from the outlet of the supply liquid reservoir 22 to the tip of the pure water. In addition, in order to investigate the decrease amount of the injected pure water, after measuring the mass of the whole medium after the injection of pure water, it was rotated at a rotational speed of 1000 rpm for 40 seconds, the analysis medium was stopped, and the sample was analyzed at 95 ° C. After holding the medium and allowing it to stand for 30 minutes, the analytical medium was removed from the rotary heating apparatus, allowed to cool to room temperature, and then the mass of the entire analytical medium was measured again. The results are shown in Tables 1 and 2 below.

  As is apparent from Table 1, pure water could be sent to the flow path of the analytical medium produced in Production Example 1 by centrifugal force due to rotation. Moreover, when the rotational speed was changed to 1000, 1250, and 1500 rpm, pure water was fed at a liquid feeding speed corresponding to the rotational speed. Further, as is apparent from Table 2, the mass reduction of the entire analysis medium was very small, and it became clear that water and vapor did not leak from the flow path even when the analysis medium was heated to 95 ° C. for a certain time. .

<Production example 2>
Analytical media having the structures shown in FIGS. 11 to 14 were manufactured under the following conditions.

  As the first substrate member 11, a polycarbonate substrate having a diameter of 3 mmφ which is the first connection port 15 and the second connection port 16 and having an outer diameter of 120 mmφ, an inner diameter of 15 mmφ and a thickness of 500 μm was molded by injection molding. In addition, the holes with a diameter of 3 mm, which are both connection ports, were formed at positions matching the supply path 31 and the return path 32 on the second substrate member 12 to be formed as described later.

  The second substrate member 12 is a polycarbonate substrate having a groove having a depth of 30 μm which is a flow path 17, a supply liquid reservoir 22, a receiving liquid reservoir 23, a supply path 31, and a return path 32, and has an outer diameter of 120 mmφ. A product with an inner diameter of 15 mmφ and a thickness of 500 μm was molded by injection molding. The shape of the flow path 17 is a flow path having a width of 200 μm and a height of 30 μm, folded back with a length of 10 mm, and having a total length of 200 mm after 10 reciprocations, and a wall between the flow paths. The width was 200 μm. Then, the zigzag portion of the flow path 17 was arranged so as to be located within a range of 40 mm to 50 mm from the center of the disk-shaped medium, and eight were arranged side by side along the circumferential direction.

  As the sealing film 33, a laminate film of modified polypropylene / silica / polycarbonate (30 μm / 1 μm / 100 μm) having a hole with a diameter of 2 mmφ serving as the sample injection port 20 was punched and molded in a size of 15 mm × 15 mm. The modified polypropylene of the laminate film becomes a sealant for bonding to the substrate.

  In order to obtain the arrangement shown in FIG. 11, both molded substrates were heated and pressure-bonded while being evenly pressurized. The pressure was 106 Pa, the temperature was 130 ° C., and heating and pressing were performed for 1 minute. Further, the sealing film 33 is adhered from above the second substrate 12 by heat sealing at 150 ° C. along the shape of the affixed portion (the portion A with dot-shaped hatching in FIGS. 11 and 12). The inner space covered with the sealing film 33 was formed, and it was stuck between the sample injection port 20 and the second connection port 16 so as to form a sticking portion Aa extending in a partition shape.

<Example 2>
The following tests were performed using the analytical medium produced in Production Example 2.

  After injecting approximately 10 μL of pure water from the sample injection port 20 through an O-ring with a microsyringe and collecting it on the left side of the internal space covered with the sealing film 33 (left side as viewed from the side direction shown in FIG. 12) In the same manner, air was injected with a microsyringe to send pure water to the first connection port 15 side. At this time, since a sticking portion Aa extending in a partition shape was formed between the sample injection port 20 and the second connection port 16, pure water was not sent to the second connection port 16 side. 12 is a part extending in the shape of a partition of the sticking part Aa, and the inner space covered with the sealing film 33 is heat sealed with a baking iron in the middle of the left and right sides of the sealing film 33. The direction of the side surface shown in FIG. It was divided into right and left sides as seen from the side. In this way, pure water was injected into all eight flow paths on the analysis medium and heat sealed.

  The test similar to Example 1 was done about the analytical medium which inject | poured the pure water as mentioned above and heat-sealed. In the test for examining the liquid feeding due to the rotation of the analysis medium, the temperature condition is the same as in Example 1 at 95 ° C., and the analysis medium rotation heating apparatus (shown schematically in FIG. 18). The heating was not performed and the temperature was 25 ° C. The results are shown in Tables 3 to 5 below.

  As is apparent from Table 3, pure water could be sent to the flow path of the analytical medium produced in Production Example 2 by centrifugal force due to rotation. Moreover, when the rotational speed was changed to 1000, 1250, and 1500 rpm, pure water was fed at a liquid feeding speed corresponding to the rotational speed.

  Further, as is apparent from Table 3, even when performed at a temperature of 25 ° C., pure water can be sent to the flow path of the analytical medium produced in Production Example 2 by the centrifugal force due to rotation. It was. In this case, the liquid feeding speed was slower than that performed under the temperature condition of 95 ° C. This decrease in the feeding speed was considered to be caused by an increase in the viscosity of pure water injected into the analysis medium by changing the temperature condition from 95 ° C. to 25 ° C.

  Further, as is apparent from Table 5, it was clarified that the decrease in the mass of the entire analysis medium was very small, and even when the analysis medium was heated to 95 ° C. for a certain time, water and vapor did not leak from the flow path. .

<Comparative Example 1>
The following tests were performed using the analytical medium produced in Production Example 2.

  That is, the test was performed in the same manner as in Example 2 except that pure water was injected into the analytical medium produced in Production Example 2 and then the sealing film 33 was not heat sealed. The results are shown in Tables 6 and 7 below.

  As a result, as shown in Table 6, when the test was performed under the temperature condition of 25 ° C., the liquid could be fed in the same manner as in Example 2 when the test was performed under the temperature condition of 25 ° C. However, when performed at a temperature of 95 ° C., the pure water injected into the analysis medium at the stage of standing at 95 ° C. for 10 minutes before rotation decreases due to evaporation, and the pure water flow is measured. I couldn't. Furthermore, as is apparent from Table 7, all of the injected pure water disappeared when exposed to 95 ° C. for at least 30 minutes under the temperature condition.

<Comparative Example 2>
Among the analytical media manufactured in Production Example 2, the following test was performed using the media without the sealing film 33 attached.

  That is, about 2.5 μL of pure water was injected from the first connection port 15 through the O-ring with the microsyringe until the supply reservoir 22 was almost full. Thereafter, both connection ports 15 and 16 were filled with an epoxy resin and sealed. This is installed in the analytical medium rotation heating apparatus (schematically shown in FIG. 18), and is rotated at a predetermined rotation speed and time as shown in Table 8 below at 25 ° C. without operating the heating means. I let you. After the elapse of a predetermined time, the rotation of the analysis medium was stopped, and the position of the tip of the supplied pure water was measured as the distance from the outlet of the supply liquid reservoir 22 to the tip of the pure water. In addition, in order to investigate the decrease amount of the injected pure water, after measuring the mass of the whole medium after the injection of pure water, it was rotated at a rotational speed of 1000 rpm for 40 seconds, the analysis medium was stopped, and the sample was analyzed at 95 ° C. After holding the medium and allowing it to stand for 30 minutes, the analysis medium was removed from the rotary heating apparatus, allowed to cool to room temperature, and then the mass of the entire analysis medium was measured again. The results are shown in Tables 8 and 9 below.

  As a result, since both the connection ports 15 and 16 were sealed with epoxy resin, the decrease in pure water injected into the analysis medium was small as shown in Table 9, but the analysis was performed as shown in Table 8. Pure water injected into the working medium could not be fed by centrifugal force.

  From the above results, it is clear that the solution in the sample evaporates and the sample concentration and the like change unless the opening of the analysis medium is sealed. In addition, if the opening is sealed with epoxy resin or the like, there is no room to accept the air that moves as the sample moves through the flow path in the analytical medium. It became clear that liquid could not be fed. On the other hand, using the analysis medium manufactured in Production Examples 1 and 2, there is room for receiving the air that moves as the sample moves in the flow path while sealing the opening of the analysis medium. It was revealed that the sample can be smoothly fed by the centrifugal force generated by the rotation of the analysis medium.

1 is an exploded perspective view showing a first embodiment of an analysis medium of the present invention. It is a partial enlarged plan view which shows the whole passage of one sample of the same medium for analysis. It is sectional drawing along the III-III arrow line of FIG. It is sectional drawing along the IV-IV arrow line of FIG. It is a top view which shows the whole medium for the analysis. It is the elements on larger scale which show the zigzag part of the flow path of the medium for the analysis. It is a top view which shows the whole medium for the analysis which has the other aspect of the zigzag part of a flow path. It is a disassembled perspective view which shows 2nd Embodiment of the medium for analysis of this invention. It is sectional drawing in the same arrangement | positioning as sectional drawing along the III-III arrow line in FIG. It is sectional drawing in the same arrangement | positioning as sectional drawing along the IV-IV arrow line in FIG. It is a disassembled perspective view which shows 3rd Embodiment of the medium for analysis of this invention. It is a partial enlarged plan view which shows the whole passage of one sample of the same medium for analysis. It is sectional drawing along the IX-IX arrow line of FIG. It is sectional drawing along the XX arrow line of FIG. It is a disassembled perspective view which shows 4th Embodiment of the medium for analysis of this invention. It is sectional drawing in the same arrangement | positioning as sectional drawing along the III-III arrow line in FIG. It is sectional drawing in the same arrangement | positioning as sectional drawing along the IV-IV arrow line in FIG. It is explanatory drawing which shows the rotation and heating method of the medium for analysis in an Example.

Explanation of symbols

10, 30 Analysis medium 11 First substrate member 11a Substrate member 12 Second substrate member 12a Sealing member 13, 14 Support hole 15 First connection port 16 Second connection port 17 Channel 17a, 17b End portion 17c Zigzag portion 18 First DESCRIPTION OF SYMBOLS 1 Sealing film 19 2nd sealing film 20 Sample injection port 21 Communication path 22 Supply liquid reservoir part 23 Receiving liquid reservoir part 31 Supply path 32 Return path 33 Sealing film 34 Communication gap 40 Analytical medium rotation heating apparatus 41 Heating means 42 Analysis media support turntable 43 Motor A, Aa
H Heater b High temperature range B Low temperature range c Electromagnetic wave

Claims (11)

A flow that communicates with the rotatable medium through the sample inlet, a supply reservoir that communicates with the sample inlet, a receiver reservoir, the supply reservoir, and the receiver reservoir. An analysis medium configured to flow a sample from the supply reservoir to the receiving reservoir through the flow path by a centrifugal force caused by rotation,
The medium is mainly composed of a substrate, and the substrate includes the flow path, a first connection port communicating with one end of the flow path, and the other end of the flow path. A second connection port is formed,
A first sealing film for sealing the first connection port and a second sealing film for sealing the second connection port are attached to the substrate surface,
Between the first sealing film and the substrate surface, the supply liquid reservoir portion including a bag-like space communicating with the first connection port is formed, and the second sealing film and the substrate surface In between, the receiving liquid reservoir portion formed of a bag-like space communicating with the second connection port is formed,
The first sealing film is formed with a sample communication port that communicates with the sample injection port, the sample injection port, and the supply liquid reservoir, and the first sealing film is supplied through the sample injection port. An analysis medium, wherein the sample communication part can be sealed after the sample is filled in the liquid reservoir. The receiving liquid reservoir is formed on the outer peripheral side of the medium with respect to the supply liquid reservoir, and the second connection port is on the outer peripheral side of the medium with respect to the first connection port. The analytical medium according to claim 1, wherein the analytical medium is formed so as to be located in the area. The medium is mainly composed of a substrate formed by joining a first substrate member and a second substrate member, and the flow path is formed between the first substrate member and the second substrate member. The analytical medium according to claim 1 or 2, wherein The medium is configured mainly with a substrate formed by joining a substrate member and a sealing member, and the flow path is formed between the substrate member and the sealing member. The analytical medium described. The analysis according to any one of claims 1 to 4, wherein the first sealing film can be sealed by welding or adhering the first sealing film to the substrate at the sample communication portion. Medium. A flow that communicates with the rotatable medium through the sample inlet, a supply reservoir that communicates with the sample inlet, a receiver reservoir, the supply reservoir, and the receiver reservoir. An analysis medium configured to flow a sample from the supply reservoir to the receiving reservoir through the flow path by a centrifugal force caused by rotation,
The medium is mainly composed of a substrate, and the substrate communicates with the supply liquid reservoir, the receiving liquid reservoir, the flow path, and the supply liquid reservoir. A first connection port and a second connection port communicating with the receiving liquid reservoir,
On the surface of the substrate, one sealing film for sealing the first connection port and the second connection port is attached,
Between the sealing film and the substrate surface, a communication gap that connects the first connection port and the second connection port is formed,
The sealing film is formed with the sample inlet, and a sample communication portion that communicates with the sample inlet and the first connection port, and the sealing film passes through the sample inlet to the supply liquid reservoir. An analysis medium characterized in that the sample communication part can be sealed after the sample is filled. The receiving liquid reservoir is formed on the outer peripheral side of the medium with respect to the supply liquid reservoir, and the second connection port is on the outer peripheral side of the medium with respect to the first connection port. The analytical medium according to claim 6, wherein the analytical medium is formed so as to be located in the area. The medium is mainly composed of a substrate formed by joining a first substrate member and a second substrate member, and the supply liquid is provided between the first substrate member and the second substrate member. The analysis medium according to claim 6 or 7, wherein a reservoir, the receiving liquid reservoir, and the flow path are formed. The medium is mainly composed of a substrate formed by joining a substrate member and a sealing member, and the supply liquid reservoir and the receiving liquid are interposed between the substrate member and the sealing member. The analysis medium according to claim 6 or 7, wherein a reservoir and the flow path are formed. The analysis medium according to any one of claims 6 to 9, wherein the sealing film can be sealed by welding or adhering the sealing film to the substrate at the sample communication portion. The flow path communicating with the supply liquid reservoir and the receiving liquid reservoir has a zigzag portion that is directed in the circumferential direction as a whole while alternately folding outward and inward along the radial direction of the medium. The medium for analysis according to any one of claims 1 to 10, wherein the medium for analysis is arranged in a temperature region in which the inner peripheral side and the outer peripheral side of the zigzag portion are different when the sample is analyzed. .

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