WO2021015145A1

WO2021015145A1 - Chip for generating thermal convection and reaction method

Info

Publication number: WO2021015145A1
Application number: PCT/JP2020/027955
Authority: WO
Inventors: 井上　直哉; 大介甲田; 秋山　博; 良彰植森; 真人齋藤
Original assignee: コニカミノルタ株式会社; 国立大学法人大阪大学
Priority date: 2019-07-25
Filing date: 2020-07-17
Publication date: 2021-01-28
Also published as: JP7549358B2; JPWO2021015145A1

Abstract

This chip 1 for generating thermal convection is provided with a rotating body 1c having formed therein a flow path 5 for thermal convection, an introduction port 50, and a supply path 10. A liquid-receiving section 51 communicating with the introduction port is formed in the rotating body, an inner surface 51a and an inner bottom surface 51b of the liquid-receiving section abut each other at an acute angle, and the supply path comprises a liquid inflow port 11a that has the base thereof arranged on the inner bottom surface of the liquid-receiving section and that opens on the inner surface. In addition, a suction passage in the supply path has a weighing space 13 and is configured so that liquid within the weighing space is separated from other liquid and supplied to the flow path for thermal convection by the centrifugal force of the rotating body. The edge of the outlet 13b of the weighing space on the flow path for thermal convection side is separated from the edge of a planar surface formed by the rotating body and opens on the planar surface. A surplus liquid storage section 15 in which liquid separated from the liquid within the weighing space by centrifugal force is stored is formed in the rotating body. A sealing agent that is a solid at room temperature is held in a sealing agent space 20.

Description

Chip for heat convection generation and reaction method

The present invention relates to a chip for generating heat convection and a reaction method.

As a gene amplification method, a polymerase chain reaction (hereinafter abbreviated as "PCR") is known. PCR is a method that can amplify a large amount of a specific DNA fragment from an extremely small amount of DNA sample in a short time, and is used not only in basic research but also in a wide range of fields from clinical genetic diagnosis to food hygiene inspection and criminal investigation. ing.
Thermal convection PCR has been proposed as a method of promoting PCR. Patent Document 1 discloses a disk-shaped microchannel chip having an annular channel for performing centrifugally accelerated thermal convection PCR.
The thermal convection generation chip described in the same document has three solution inlets (“accepting part 121” in the same document), one is a sample liquid, one is a PCR liquid, and one is an evaporation suppressing liquid ( Mineral oil) is introduced. A micro flow path (“pull passage 122” in the same document) extends from there to form a V-shaped flow path, and a space region for weighing (“first region 122a” in the same document) for storing a solution on the downstream side. ). When the volumes of the sample solution region and the PCR solution region are combined, the volume becomes the same as that of the annular flow path (“heat convection flow path 11” in the same document). Each liquid introduced from the introduction port (“reception section 121” in the same document) enters the microchannel by capillarity and fills the weighing space area. At this time, by centrifuging, the liquid in the weighing region is transferred to the annular flow path starting from the valley of the V-shaped structure, and the surplus is returned to the introduction port side so that the liquid of the required capacity for the annular flow path is released. Be supplied. Further, by arranging a plurality of V-shaped channels for weighing and arranging an annular channel downstream of each branch, it is possible to detect a plurality of types of genes in one sample.

International Publication No. 2015/170753

However, in the above-mentioned prior art, it is further improved to control the liquid in the microchannel in a planned region by flowing and stabilizing it at each stage from the introduction of the solution to the execution of the centrifugally accelerated thermal convection PCR. There was room, and there was a risk of excess or deficiency in the amount of liquid supplied to the heat convection flow path.

The present invention has been made in view of the above problems in the prior art, and an object of the present invention is to improve the accuracy of the amount of liquid supplied to the heat convection flow path.

The first aspect of the present invention includes a rotating body in which a heat convection flow path, an introduction port and a supply path are formed, and the liquid introduced into the introduction port is supplied to the heat convection flow path by the supply path. A chip for generating heat convection in which heat is convected in the flow path for heat convection. A liquid receiving portion communicating with the introduction port is formed on the rotating body, and the inner side surface and the inner bottom surface of the liquid receiving portion have sharp angles. The supply path has a liquid inflow port that is open on the inner side surface by arranging the bottom surface on the inner bottom surface of the liquid receiving portion.

A second aspect of the present invention includes a rotating body in which a heat convection flow path, an introduction port, and a supply path are formed, and supplies the liquid introduced into the introduction port to the heat convection flow path by the supply path. A chip for generating heat convection in which heat is convected in the heat convection flow path, and a liquid receiving portion communicating with the introduction port is formed in the rotating body, and the supply path allows the liquid in the liquid receiving portion to be formed. The suction passage has a suction passage for sucking by a capillary phenomenon, the suction passage has a weighing space, and the suction passage has a liquid in the weighing space in another region due to the centrifugal force when the rotating body is rotated. It is configured to be separated from the liquid inside and supplied to the heat convection flow path, and the outlet of the weighing space on the heat convection flow path side is separated from the edge of the plane formed by the rotating body at the edge of the outlet. And it is open to the plane.

A third aspect of the present invention includes a rotating body in which a heat convection flow path, an introduction port, and a supply path are formed, and a liquid introduced into the introduction port is introduced into the heat convection flow path by the supply path. A chip for generating heat convection that is supplied and heat convected in the flow path for heat convection. A liquid receiving portion communicating with the introduction port is formed in the rotating body, and the supply path is a liquid in the liquid receiving portion. The suction passage has a weighing space, and the suction passage has a liquid in the weighing space due to the centrifugal force when the rotating body is rotated. A surplus liquid storage portion is formed in the rotating body so as to be separated from the liquid in the region and supplied to the heat convection flow path, and the liquid separated from the liquid in the weighing space is stored by the centrifugal force. Has been done.

A fourth aspect of the present invention includes a rotating body in which a heat convection flow path, an introduction port, and a supply path are formed, and a liquid introduced into the introduction port is introduced into the heat convection flow path by the supply path. A heat convection generating chip that is supplied and heat convected in the heat convection flow path, and communicates with the rotating body to a connection portion of the supply path with the heat convection flow path separately from the supply path. The sealant space is formed, the sealant space holds the solid sealant at room temperature, and the sealant is closed in order to block the heat convection flow path to which the liquid is supplied by the supply path. The sealing agent in the space for stopping agent is heated and melted, and is transferred to the connecting portion by the centrifugal force when the rotating body is rotated so that it can be filled.

According to the first aspect of the present invention, the liquid can be reliably introduced into the supply path and the supply shortage can be prevented, so that the accuracy of the liquid supply amount to the heat convection flow path can be improved.
According to the second aspect of the present invention, the unintended leakage of the liquid from the weighing space for weighing the supply amount to the heat convection flow path side can be prevented, so that the accuracy of the liquid supply amount to the heat convection flow path can be improved. Can be improved.
According to the third aspect of the present invention, since the surplus liquid is held in the surplus liquid storage portion, the accuracy of the amount of liquid supplied to the heat convection flow path can be improved.
According to the fourth aspect of the present invention, the heat convection flow path can be blocked by the existing sealant in the heat convection generation chip by heating and rotating at an appropriate timing. The accuracy of the liquid supply to the can be maintained.

It is a top perspective view of the heat convection generation chip which concerns on one Embodiment of this invention. It is a top perspective view of the heat convection generation chip which concerns on one Embodiment of this invention, and shows the state which the upper lid is removed. It is a bottom perspective view of the heat convection generation chip which concerns on one Embodiment of this invention. It is a bottom perspective view of the heat convection generation chip which concerns on one Embodiment of this invention, and shows the state which the bottom lid is removed. It is a top view of the heat convection generation chip which concerns on one Embodiment of this invention, and shows the state which the upper lid is removed. It is a bottom view of the heat convection generation chip which concerns on one Embodiment of this invention, and shows the state which the bottom lid is removed. It is a central sectional view of the chip for heat convection generation which concerns on one Embodiment of this invention. It is a partially enlarged top perspective view of the heat convection generation chip which concerns on one Embodiment of this invention, and shows the state which removed the upper lid. It is a partially enlarged top perspective view of the heat convection generation chip which concerns on one Embodiment of this invention, and shows the state which removed the upper lid. It is a perspective view seen from the angle different from FIG. It is a partially enlarged top view of the heat convection generation chip which concerns on one Embodiment of this invention, and shows the state which removed the upper lid. It is a partially enlarged bottom perspective view of the heat convection generation chip which concerns on one Embodiment of this invention, and shows the state which removed the bottom lid.

An embodiment of the present invention will be described below with reference to FIGS. 1 to 11. The following is an embodiment of the present invention and does not limit the present invention.

As shown in FIG. 1 and the like, the heat convection generation chip 1 of the present embodiment is a disk-shaped microchannel chip, and has a rotating body 1c having a structure in which a disk 1a and an upper cylinder 1b are connected by a coaxial A as a main body. ..
The disk 1a includes a core substrate 2, a ring-shaped top lid 3 into which an upper cylinder 1b can be inserted, and a bottom lid 4. The core substrate 2 has an upper surface 2A and a lower surface 2B. A groove forming a flow path or the like is formed in the core substrate 2, the upper surface opening of the groove is closed by the upper lid 3, and the lower surface opening is closed by the bottom lid 4, so that the flow path and the air passage are formed. However, the air port 100 provided on the upper surface is open.
A notch 99 that is held during rotation or the like is formed on the outer edge of the disk 1a.

Further, the heat convection generation chip 1 is a 12-channel PCR microchannel chip, and 12 sets of heat convection channels 5 and a supply path 10 for supplying a solution to the disk 1a are central angles. Is divided into 12 equal parts and arranged in a divided range. The heat convection flow path 5 is a flow path formed in an annular shape.
The upper end opening of the upper cylinder 1b is an introduction port 50. The space below the introduction port 50 is a liquid receiving unit 51 communicating with the introduction port 50. The bottom surface of the liquid receiving portion 51 is composed of the upper surface of the bottom lid 4.
The liquid introduced into the introduction port 50 is distributed and supplied to the heat convection flow paths 5 of 12 through the supply passages 10 of 12, and heat convection is performed in each heat convection flow path 5.

The configuration of the supply path 10 is as follows in the order from the most upstream liquid receiving portion 51 to the most downstream heat convection flow path 5.
That is, the supply passage 10 has a configuration in which the first passage 11 whose upstream end is connected to the liquid receiving portion 51, then the second passage 12, the weighing space 13, and finally the introduction chamber 14 are connected to each other. The downstream end is connected to the heat convection flow path 5.
The liquid receiving portion 51, the first passage 11, the second passage 12, the weighing space 13, the introduction chamber 14, and the heat convection flow path 5 are continuous in this order.
The first passage 11, the second passage 12, and the weighing space 13 correspond to a suction passage that sucks the liquid in the liquid receiving portion 51 by a capillary phenomenon. Therefore, the liquid in the liquid receiving portion 51 passes through the first passage 11 and the second passage 12 due to the capillary phenomenon, and is filled in the weighing space 13.
The first passage 11 is a flow path that transitions outward in the radial direction. The second passage 12 is a flow path that transitions inward in the radial direction. The weighing space 13 is a flow path that transitions outward in the radial direction. The term "diameter direction" refers to the radial direction centered on the rotation center axis A of the rotating body 1c.
Further, the introduction chamber 14 is arranged radially outward from the weighing space 13. The heat convection flow path 5 is arranged radially outward from the introduction chamber 14.

An air introduction port 13a is opened on the upper surface of the upstream end of the weighing space 13. The outlet 13b is open on the upper surface of the downstream end of the weighing space 13.
In the stationary state of the rotating body 1c, the liquid that fills the weighing space 13 due to the capillary phenomenon stops in the form of stretching the surfaces at the air introduction port 13a and the outlet 13b.

Next, a flow path and a space other than the series system from the liquid receiving portion 51 to the heat convection flow path 5 will be described.
The downstream end of the first passage 11 and the upstream end of the second passage 12 are connected to the upstream end of the surplus liquid storage portion 15.
A dam portion 15a is formed at the upstream end of the surplus liquid storage portion 15.
The surplus liquid storage portion 15 bypasses the radial outer side of the heat convection flow path 5 and extends in an arc shape so as to straddle the outermost radial end 15e of the surplus liquid storage portion 15. The downstream end of the surplus liquid storage portion 15 is connected to the air passage 16a in the radial direction from the outermost radial end 15e. The air passage 16a, the filter installation chamber 17, and the air passage 18 are continuous in this order.
The sealant space 20 communicates with the introduction chamber 14 via the sealant supply path 19. The sealing agent space 20 is arranged inward in the radial direction from the introduction chamber 14 in order to enable the supply of the sealing agent by centrifugal force.
Further, the air introduction port 13a of the weighing space 13 and the radial inner end of the sealing agent space 20 are connected to the air passage 16b. The air passage 16b, the filter installation chamber 17, and the air passage 18 are continuous in this order.
The portion of the air passage 18 that is not blocked by the upper lid 3 is the air port 100. In order to prevent liquid leakage from the air port 100, the air port 100 is installed at a position radially inward from the weighing space 13 and the sealing agent space 20. In the filter installation chamber 17, a filter having a property of allowing air to pass through and not allowing liquid to pass through is installed.

With the above configuration, the liquid in the weighing space 13 is separated from the liquid in the other regions (11, 12) and supplied to the heat convection flow path 5 by the centrifugal force when the rotating body 1c is rotated. .. At this time, the introduction of air from the air passage 16b promotes the separation of the liquid in the weighing space 13 and the liquid in the second passage 12.

As shown in FIG. 7, the inner side surface 51a and the inner bottom surface 51b of the liquid receiving portion 51 are in contact with each other at an acute angle.
The supply path 10 has a liquid inflow port 11a having a bottom surface arranged on the inner bottom surface 51b of the liquid receiving portion 51 and opening to the inner side surface 51a. The inner bottom surface 51b is composed of the upper surface of the bottom lid 4. The liquid inflow port 11a is an upstream end opening of the first passage 11. Since the inner bottom surface of the first passage 11 is also composed of the upper surface of the bottom lid 4, the inner bottom surface 51b of the liquid receiving portion 51 and the inner bottom surface of the first passage 11 are at the same height level. Therefore, the bottom of the liquid inlet 11a is arranged on the inner bottom surface 51b of the liquid receiving portion 51.
As described above, the surface (inner side surface 51a) through which the liquid inflow port 11a opens is continuous in contact with the inner bottom surface 51b of the liquid receiving portion 51 at an acute angle, and adjacent surfaces are in front of the liquid inflow port 11a. By forming a narrow space that creates an acute angle, a capillary phenomenon is generated there, and the liquid in the liquid receiving portion 51 can be quickly flowed into the liquid inlet 11a.
As a result, the introduction of the liquid into the supply path 10 is quick and reliable, and the supply shortage can be prevented, so that the accuracy of the liquid supply amount to the heat convection flow path 5 can be improved.

In the present embodiment, the liquid inlets 11a of the plurality of sets of supply passages 10 are similarly opened to the inner side surface 51a of one liquid receiving portion 51. This makes it possible to distribute the solution from one liquid receiving unit 51.
Further, the central axis A of the introduction port 50 and the liquid receiving portion 51 is arranged on the rotating central axis A of the rotating body 1c, and the inner side surface 51a of the liquid receiving portion 51 is arranged equidistant from the rotating central axis A. .. Further, at least the lower portion of the inner side surface 51a of the liquid receiving portion 51 is formed by a tapered surface that expands toward a lower position, and the liquid inflow port 11a opens in the tapered surface. As a result, it is easy to distribute the liquid from one liquid receiving unit 51 to a plurality of sets of supply paths 10 evenly and simultaneously.

As shown in FIG. 10, the outlet 13b on the heat convection flow path 5 side of the weighing space 13 opens in the plane 13d with the edge of the outlet 13b separated from the edge of the plane 13d formed by the rotating body 1c. .. As a result, the surrounding surface surrounding the outlet 13b is arranged on a plane including the opening surface of the outlet 13b. Therefore, the liquid that has reached the outlet 13b is prevented from coming into contact with the outer surface of the outlet 13b, and the liquid is prevented from leaking from the outlet 13b. If there is a liquid leaking from the outlet 13b, the liquid will be introduced into the heat convection flow path 5, so that the weighing accuracy will drop. Therefore, the weighing accuracy is improved by preventing the liquid from leaking from the outlet 13b.
Similarly, the air introduction port 13a of the weighing space 13 also opens to the plane 13c with the edge of the air introduction port 13a separated from the edge of the plane 13c formed by the rotating body 1c.
Therefore, it is possible to prevent the liquid from leaking from the air inlet 13a, and the weighing accuracy is improved.
As described above, since only the liquid having a capacity that fills the weighing space 13 is supplied to the heat convection flow path 5, the accuracy of the liquid supply amount to the heat convection flow path 5 can be improved.
In the present embodiment, the plane 13d where the outlet 13b opens and the plane 13c where the air introduction port 13a opens are both upward horizontal planes, but in order to obtain the effect of preventing the liquid from leaking out. The angle of the opening surface of the outlet 13b and the air introduction port 13a may be any angle. For example, the same effect can be obtained by opening the outlet 13b and the air inlet 13a in a vertical plane or a downward horizontal plane.

The surplus liquid storage unit 15 is a space in which the liquid separated from the liquid in the weighing space 13 (the liquid in the passages 11 and 12) is stored by the centrifugal force generated by the rotation of the rotating body 1c. Exhaust through the air passage 16a promotes the inflow of liquid into the surplus liquid storage unit 15. Since the excess liquid storage portion 15 extends from the same position in the radial direction to the outside as the heat convection flow path 5, the centrifugal force due to the rotation of the rotating body 1c acts equal to or more than the heat convection flow path 5. When the centrifugal force due to the rotation of the rotating body 1c is working, the liquid is easily held around the outermost radial end 15e of the surplus liquid storage portion 15, and the air passage at the downstream end of the surplus liquid storage portion 15. By 16b, it is difficult for the liquid to move.

The dam portion 15a is formed so as to project upward from the bottom surface of the excess liquid storage portion and cross between both side surfaces. The dam portion 15a has an action of blocking the liquid that is about to flow out from the excess liquid storage portion 15. The dam portion 15a is provided inward in the radial direction from the outermost end 15e in the radial direction. The dam portions 15a may be provided in duplicate or more.
The cross section of the flow path of the excess liquid storage portion 15 has rounded corner portions 15R. The rounded corner portion 15R is a corner portion formed by the dam portion 15a and both side surfaces in the dam portion 15a. Other than that, the corners formed by the top surface and both side surfaces of the surplus liquid storage portion 15 are rounded corners 15R. This is because the corners formed by the top lid 3 or the bottom lid 4 and the core substrate 2 cannot be rounded due to molding reasons.
Sharp corners cause capillarity. Since the corner portion 15R is rounded, it is possible to suppress backflow due to the capillary phenomenon. Therefore, it is possible to prevent the surplus liquid that has once flowed into the surplus liquid storage unit 15 from flowing back toward the first passage 11 and the second passage 12. Further, since the excess liquid storage portion 15 is suppressed from flowing due to the capillary phenomenon, the liquid of the liquid receiving portion 51 passes through the first passage 11 and the second passage 12 due to the capillary phenomenon in the stationary state of the rotating body 1c, and is a weighing space. When introduced into 13, the inflow of the liquid into the surplus liquid storage unit 15 is also suppressed. Therefore, there is an effect of preventing a situation in which the liquid is unevenly distributed to the 12 channels due to the uneven inflow of a large amount of the liquid into a part of the surplus liquid storage portion 15 out of the 12 channels.
By providing the dam portion 15a, it is possible to provide a rounded corner portion 15R not only on the top surface side but also on the bottom surface side, so that the liquid to flow back along the corner portion is rounded. It is blocked by either corner 15R. The radius of curvature of the corner portion 15R is preferably 0.1 mm or more.
As described above, since the surplus liquid is reliably held in the surplus liquid storage unit 15, the accuracy of the amount of liquid supplied to the heat convection flow path 5 can be improved.

The contents of the experiment for investigating the relationship between the radius of curvature of the corner portion (15R) of the cross section of the flow path and the flow distance following the corner portion are disclosed below.
A required number of grooves having a depth of 0.5 mm, a width of 2.5 mm, and a length of 30.0 mm were formed on the upper surface of the resin plate, and these grooves were used as an experimental flow path. Therefore, the experimental flow path has a corner formed by the bottom surface and one side surface, and a corner portion formed by the bottom surface and the other side surface, and the upper surface is open.
Five types of flow paths for experiments were prepared in which the radius of curvature R of the two corners was 0.0, 0.05, 0.1, 0.2, and 0.3.
The resin plate was kept horizontal, and the same amount (5 μliter) of the colored solution was injected into the longitudinal end of each experimental flow path and observed. Then, the colored solution moved so as to extend along the corners on both sides of the flow path. The moving distance 90 seconds after the injection was as follows.
22.3 mm in the flow path of R = 0.0
In the flow path of R = 0.05, 14.1 mm
In the flow path of R = 0.1, 7.0 mm
In the flow path of R = 0.2, 3.2 mm
2.8 mm in the flow path of R = 0.3

Further, the width of the flow path of the dam portion 15a, the front portion 15b, and the rear portion 15c thereof is widened with respect to the storage portion 15d further downstream in the inflow direction from the rear portion 15c. As a result, the backflow path that follows the corners becomes longer, so that it becomes difficult for the liquid in the surplus liquid storage portion 15 to flow back and flow out toward the first passage 11 and the second passage 12.
Since the core substrate 2 penetrates vertically in the flow path cross sections of the front portion 15b and the rear portion 15c, the rounded corner portions 15R are not provided. However, the vertical corners 15U in the front 15b and the rear 15c are rounded. The axis of the radius of curvature of the corner portion 15U is parallel to the rotation center axis A of the rotating body 1c. In addition to the rounded corners 15R, the presence of the rounded corners 15U further suppresses the flow due to the capillary phenomenon in the excess liquid storage portion 15, and prevents unintended inflow and backflow. ..
The dam portion 15a and the storage portion 15d are provided with rounded corner portions 15R.

As described above, in the rotating body 1c, apart from the supply path 10, a connecting portion of the supply path 10 with the heat convection flow path 5, that is, a sealing agent space 20 communicating with the introduction chamber 14 is formed. There is.
Paraffin or the like is pre-loaded in the sealing agent space 20 as a solid sealing agent at room temperature, and then the top lid 3 and the bottom lid 4 are attached to the core substrate 2.
As described above, after the liquid from the liquid receiving portion 51 is introduced into the heat convection flow path 5 by the introduction of the liquid into the weighing space 13 by the capillary phenomenon and the subsequent rotation of the rotating body 1c, the following is performed. The heat convection flow path 5 is closed.
That is, the sealant in the sealant space 20 is heated, melted, and transferred to the connection portion (introduction chamber 14) by the centrifugal force when the rotating body 1c is rotated and filled. At this time, the introduction of air from the air passage 16b promotes the outflow of the sealant from the sealant space 20.
As a result, the introduction chamber 14 is filled with the sealant, and the heat convection flow path 5 is closed. Therefore, the introduction chamber 14 is adjacent to the heat convection flow path 5 downstream of the weighing space 13 and is also a space for closing the inlet of the heat convection flow path 5 by filling with a sealing agent. Since the heat convection flow path 5 is blocked, leakage due to evaporation of the liquid in the heat convection flow path 5 can be prevented.
As described above, by heating and rotating at an appropriate timing, the heat convection flow path 5 can be blocked by the existing sealant in the heat convection generation chip 1, so that the liquid to the heat convection flow path 5 can be closed. The accuracy of the supply amount can be maintained.

The procedure from liquid introduction to thermal convection reaction will be explained again as follows.
Primer DNA and probe DNA are arranged in advance in each heat convection flow path 5 in a dried state. This corresponds to various DNAs (bacterial species).
First, a sample which is a collected biological substance is mixed in a sample container containing a predetermined reaction reagent solution to prepare a solution containing the sample and the reaction reagent. As the sample container, a container in which the lower end opening is laminated with aluminum foil or the like is used.
In the center of the liquid receiving portion 51, an opening protrusion 52 having a sharp upper portion is provided.
The sample container is inserted into the liquid receiving portion 51 from the introduction port 50, the laminate is broken by the opening protrusion 52 to open the sample container, and the solution containing the sample and the reaction reagent in the sample container is received as a liquid. Introduce to section 51.

Then, as described above, the solution is introduced up to the weighing space 13 due to the weight of the solution and the capillary phenomenon.
After the weighing space 13 is filled with the same solution, the rotating body 1c is rotated to supply only the solution in the weighing space 13 to the heat convection flow path 5.
Since the same flow occurs in the supply passage 10 and the heat convection flow path 5 of each set, the solution is distributed and supplied to the heat convection flow paths 5 of a plurality of sets.
Next, after closing the heat convection flow path 5 with the above-mentioned encapsulant, the solution is thermally convected in each heat convection flow path 5 under each predetermined condition to carry out a polymerase chain reaction or a reverse transcription polymerase chain reaction. I do.

As described above, according to the heat convection generation chip of the present embodiment, the accuracy of the amount of liquid supplied to the heat convection flow path 5 can be improved.
Further, it is easy to automate the work from loading the sample container into the introduction port 50 to the heat convection reaction, and the reactions in the plurality of heat convection flow paths 5 can be carried out at the same time, so that the workability is improved. Can be done. The sample container may be manually loaded into the introduction port 50, but by letting the machine perform the loading, more stable control of the work operation and a wider range of automation can be realized. In particular, the heat convection generation chip 1 already has a solid sealant, and the sample container has one loading part (introduction port 50), and after the sample container is loaded into the introduction port 50, it rotates. Since it can be carried out by rotating the body 1c and controlling the temperature of each part, workability is improved, and the work load of the machine due to automation is small and automation is easy.
In the heat convection generation chip of the above embodiment, the number of channels (the number of heat convection channels) is set to 12, but the number of channels is not limited and may be more than 12 channels or less than 12 channels. It can be implemented by selecting the number of channels. This makes it possible to support a wide range of required channels.

The present invention can be used for a chip for generating heat convection and a reaction method.

1 Chip for heat convection generation 1a Disk 1b Upper cylinder 1c Rotating body 2 Core substrate 2A Upper surface of core substrate 2B Lower surface of core substrate 3 Top lid 4 Bottom lid 5 Heat convection flow path 10 Supply path 11 First passage 11a Liquid inlet 12 2nd passage 13 Weighing space 13a Air introduction port 13b Outlet

13c Flat surface

13d Flat surface 14 Introduction chamber 15 Surplus liquid storage part

15R Square part

15a Dam part 15b Front part of dam part 15c Rear part of dam part 15d Storage part 15e Surplus liquid storage part Radial outermost end

16a Air passage

16b Air passage 17 Filter installation chamber 18 Air passage 19 Sealant supply path 20 Sealing agent space 50 Inlet port 51 Liquid receiving part 51a Inner side surface 51b Inner bottom surface 52 Opening protrusion 99 Notch 100 Air port A Rotation center axis

Claims

A rotating body having a heat convection flow path, an introduction port, and a supply path is provided, and the liquid introduced into the introduction port is supplied to the heat convection flow path by the supply path, and heat is generated in the heat convection flow path. A chip for generating heat convection to be convected,
A liquid receiving portion communicating with the introduction port is formed on the rotating body, and the liquid receiving portion is formed.
The inner side surface and the inner bottom surface of the liquid receiving portion are in contact with each other at an acute angle.
The supply path is a heat convection generation chip having a liquid inlet having a bottom arranged on the inner bottom surface of the liquid receiving portion and opened on the inner side surface.
A plurality of sets of the heat convection flow path and the supply path for supplying the liquid to the heat convection flow path are formed in the rotating body.
The heat convection generation chip according to claim 1, wherein the liquid inlets of a plurality of sets of the supply passages are provided by opening on the inner side surface of one liquid receiving portion.
The central axis of the introduction port and the liquid receiving portion is arranged on the rotation central axis of the rotating body.
The inner surface of the liquid receiving portion is arranged equidistant from the rotation center axis.
At least the lower portion of the inner surface of the liquid receiving portion is formed by a tapered surface that expands toward a lower position.
The heat convection generation chip according to claim 2, wherein the liquid inlet is opened on the tapered surface.
A rotating body having a heat convection flow path, an introduction port, and a supply path is provided, and the liquid introduced into the introduction port is supplied to the heat convection flow path by the supply path, and heat is generated in the heat convection flow path. A chip for generating heat convection to be convected,
A liquid receiving portion communicating with the introduction port is formed on the rotating body, and the liquid receiving portion is formed.
The supply path has a suction passage for sucking the liquid in the liquid receiving portion by a capillary phenomenon.
The suction passage has a weighing space and has a weighing space.
The suction passage is configured so that the liquid in the weighing space is separated from the liquid in the other region and supplied to the heat convection flow path by the centrifugal force when the rotating body is rotated.
The outlet on the heat convection flow path side of the weighing space is a heat convection generation chip that opens to the plane with the edge of the outlet separated from the edge of the plane formed by the rotating body.
A rotating body having a heat convection flow path, an introduction port, and a supply path is provided, and the liquid introduced into the introduction port is supplied to the heat convection flow path by the supply path, and heat is generated in the heat convection flow path. A chip for generating heat convection to be convected,
A liquid receiving portion communicating with the introduction port is formed on the rotating body, and the liquid receiving portion is formed.
The supply path has a suction passage for sucking the liquid in the liquid receiving portion by a capillary phenomenon.
The suction passage has a weighing space and has a weighing space.
The suction passage is configured so that the liquid in the weighing space is separated from the liquid in the other region and supplied to the heat convection flow path by the centrifugal force when the rotating body is rotated.
A chip for generating heat convection in which a surplus liquid storage portion for storing a liquid separated from the liquid in the weighing space by the centrifugal force is formed on the rotating body.
The heat convection generation chip according to claim 5, wherein the cross section of the flow path of the excess liquid storage portion has rounded corners.
The heat convection generation chip according to claim 5 or 6, wherein the excess liquid storage portion has a dam portion that projects upward from the bottom surface and crosses between both side surfaces.
The heat convection generation chip according to claim 7, wherein the corners formed by the dam portion and the side surfaces on both sides are rounded.
A rotating body having a heat convection flow path, an introduction port, and a supply path is provided, and the liquid introduced into the introduction port is supplied to the heat convection flow path by the supply path, and heat is generated in the heat convection flow path. A chip for generating heat convection to be convected,
In the rotating body, a space for a sealing agent is formed in addition to the supply path, which communicates with the connection portion of the supply path with the heat convection flow path.
A solid sealant is held at room temperature in the sealant space,
When the rotating body is rotated by heating and melting the sealing agent in the sealing agent space in order to block the heat convection flow path to which the liquid is supplied by the supply path. A chip for generating heat convection that is transferred to the connection portion and can be filled by centrifugal force.
Using the heat convection generation chip according to claim 3,
A solution containing a sample and a reaction reagent is introduced into the liquid receiving portion through one of the inlets, the solution is distributed and supplied to a plurality of sets of the heat convection channels, and heat is generated in each of the heat convection channels. A reaction method in which a polymerase chain reaction or a reverse transcription polymerase chain reaction is carried out by convection.