JP2011152052A - Biochip - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a biochip capable of applying the stable thermal cycle to a sample even when air bubbles exist in a chip. <P>SOLUTION: The biochip 100 comprises: a first cavity 10 filled with oil; a second cavity 20 which is adjacent to the first cavity 10 and filled with the oil; and an opening and closing mechanism 30 which is operated based on the gravity to communicate or intercept the first cavity 10 and the second cavity 20. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a biochip.

  In recent years, PCR (Polymerase Chain Reaction), which amplifies an extremely small amount of DNA in a short time, has been used in the medical field such as gene diagnosis and gene therapy, agricultural and livestock field such as breeding and breeding, food field, and crime investigation Has been developed in the field. In the PCR method, in general, a test solution (PCR reaction solution) is prepared by mixing a sample DNA and a reagent such as DNA polymerase, template DNA, primer DNA, and dNTP. This is done by raising and lowering the temperature (thermal cycling).

  On the other hand, the development of the PCR method requires an apparatus for performing the thermal cycle efficiently. Conventionally, multiple incubators set at different temperatures are prepared, and the reaction vessel (tube) containing the test solution is moved between these incubators to perform a thermal cycle, or the temperature of one thermal block A method of adjusting the temperature to a plurality of different temperatures and performing a thermal cycle has been adopted. However, these methods have a limit in reducing the time required for the thermal cycle.

  Therefore, for example, Patent Document 1 proposes a chip and a thermal cycler that efficiently perform thermal cycling of a sample by a method in which a small amount of reaction liquid is moved by gravity in a container filled with oil that is immiscible with the reaction liquid. ing. And the chip | tip disclosed by patent document 1 is filled with the oil and the reaction liquid inside, and the droplet of the reaction liquid is moved between the temperature control areas of the thermal cycler based on the action of gravity in the oil. By doing so, a desired thermal cycle can be performed.

JP 2009-136250 A

  However, the chip disclosed in Patent Document 1 is filled with oil and reaction liquid in a sealed space. For this reason, when performing an operation of filling them, there are cases where the bubbles are also filled together. Moreover, even if it can be filled without mixing air bubbles, dissolved gas contained in the filled reaction liquid or oil may cause air bubbles in the container.

  When a thermal cycler of a reaction solution is performed by a thermal cycler of a method in which the reaction solution is moved by gravity in the chip (hereinafter, sometimes referred to as “elevating type” in this specification), bubbles are generated in the chip. If there is, not only the reaction solution but also the bubbles move in the chip based on the action of gravity. For this reason, bubbles may interfere with the movement of the droplets of the reaction solution, or the movement of the bubbles may cause an unnecessary flow of oil in the chip, thereby disturbing temperature control in the chip. That is, when performing a thermal cycle with an elevating type apparatus, if bubbles exist in the chip, it may not be possible to perform a desired thermal cycle on the reaction solution.

  One of the objects according to some embodiments of the present invention is to provide a biochip capable of performing a stable thermal cycle on a sample using a thermal cycler even when bubbles are present in the chip. is there.

  SUMMARY An advantage of some aspects of the invention is to solve at least a part of the problems described above, and the invention can be implemented as the following aspects or application examples.

[Application Example 1]
One aspect of the biochip according to the present invention is:
A first cavity filled with oil;
A second cavity adjacent to the first cavity and filled with the oil;
An opening / closing mechanism that operates based on gravity, and that connects or blocks the first cavity and the second cavity;
Have

  According to such a biochip, when a liquid exists in the biochip, the bubbles can be collected in either the first cavity or the second cavity. Therefore, it is possible to prevent bubbles from being present in the other cavity. Thereby, for example, when oil and a PCR reaction solution are filled in a biochip, a stable thermal cycle can be applied to the PCR reaction solution in the other cavity.

[Application Example 2]
In application example 1,
The opening / closing mechanism may include a movable plug that opens or closes the space between the first cavity and the second cavity.

  According to the biochip of this application example, when air bubbles are present in the biochip, the air bubbles can be reliably collected in either the first cavity or the second cavity.

[Application Example 3]
In application example 1,
The opening / closing mechanism may include a movable plug that opens or closes the space between the first cavity and the second cavity.

  In the biochip of this application example, when bubbles exist in the biochip, the bubbles can be reliably collected in the second cavity.

[Application Example 4]
In any one of Application Examples 1 to 3,
The first cavity may have a shape whose longitudinal direction is a direction connecting the first cavity and the second cavity.

  According to the biochip of this application example, it has the characteristics of the biochip of the above application example, and it is easy to provide different temperature regions in the chip.

[Application Example 5]
In any one of Application Examples 1 to 4,
A partition member is formed in the first cavity,
In the first cavity, a path through which the oil can circulate around the partition member can be provided.

  According to the biochip of this application example, when a bubble exists in the biochip, a passage through which the bubble moves in the first cavity can be defined. Thereby, for example, when the biochip is filled with droplets of oil and PCR reaction liquid and bubbles are present, the bubble passage and the droplet passage can be separated. In the cavity, it is possible to suppress the movement of the droplets from being hindered by the bubbles.

[Application Example 6]
One aspect of the biochip according to the present invention is:
A first cavity filled with oil;
A second cavity that communicates with the first cavity via a communication path and is filled with the oil;
A first barrier formed between the first cavity and the communication path and protruding from an inner wall surface defining the first cavity and the communication path;
A direction parallel to the first barrier is formed between the second cavity and the communication path, and is opposite to a direction in which the first barrier protrudes from an inner wall surface defining the second cavity and the communication path. A second barrier formed protruding in the direction of
Have
The first cavity, the communication path, and the second cavity are continuous closed spaces,
When the second cavity is viewed from a direction perpendicular to the first barrier and the second barrier through the communication path from the first cavity side, the second cavity is the first barrier and It is shielded by the second barrier.

  According to such a biochip, even when bubbles exist in the biochip, the bubbles are collected in the second cavity by the action of the first barrier and the second barrier, and the bubbles are collected in the first cavity. It can be prevented from returning into the tee. Thereby, for example, when oil and PCR reaction liquid droplets are filled in a biochip and bubbles are present, a stable thermal cycle is applied to the droplets in the first cavity. be able to.

The schematic diagram of the cross section of the biochip 100 of embodiment. The disassembled perspective view which shows typically the thermal cycler 200 of embodiment. The perspective view which shows typically the thermal cycler 200 of embodiment. The figure which shows typically a mode that the biochip 100 of embodiment is loaded in the thermal cycler 200, and rotates. The partial cross section perspective view which shows typically the biochip 110 of embodiment. The schematic diagram of the cross section of the biochip 110 of embodiment. The schematic diagram of the cross section of the biochip 120 of embodiment. The schematic diagram of the cross section of the biochip 130 of embodiment. The figure which shows typically a mode that the biochip 130 of embodiment is loaded in the thermal cycler 200, and rotates. The schematic diagram of the cross section of the biochip 140 of embodiment. The figure which shows typically a mode that the biochip 140 of embodiment is loaded in the thermal cycler 200, and rotates. The schematic diagram of the cross section of the biochip 150 of embodiment. The figure which shows typically a mode that the principal part of the biochip of embodiment rotates. The figure which shows typically the principal part of the biochip of embodiment. The figure which shows typically a mode that the biochip 150 of embodiment is loaded in the thermal cycler 200, and rotates. The figure which shows typically a mode that the biochip 150 of embodiment is loaded in the thermal cycler 200, and rotates. The figure which shows typically a mode that the biochip 150 of embodiment is loaded in the thermal cycler 200, and rotates.

  Preferred embodiments of the present invention will be described below with reference to the drawings. In addition, the following embodiment demonstrates an example of this invention. Therefore, this invention is not limited to the following embodiment, The various modifications implemented in the range which does not change a summary are also included. Note that not all the configurations described in the following embodiments are essential constituent requirements of the present invention.

1. 1. First embodiment 1.1. Biochip FIG. 1 is a schematic cross-sectional view of a biochip 100 of the present embodiment. The biochip 100 of this embodiment includes a first cavity 10, a second cavity 20, and an opening / closing mechanism 30.

  The biochip 100 is a biochip used for a thermal cycler 200 (described later) and has an outer shape that can be loaded into a mounting hole 220 formed in the rotating portion 210 of the thermal cycler 200. Therefore, the outer shape of the biochip 100 can have an arbitrary shape according to the shape of the mounting hole 220 of the rotating unit 210. The size of the biochip 100 is not particularly limited, but considering at least one of the amount of liquid such as oil to be filled, the thermal conductivity, the shape of the cavity formed inside, and the ease of handling. Selected.

  The material of the biochip 100 is not particularly limited, and examples thereof include inorganic materials (for example, single crystal silicon, Pyrex glass (Pyrex is a registered trademark)), and organic materials (for example, resins such as polycarbonate and polypropylene). The composite material may be used. When using the biochip 100 as a reaction vessel (reaction chip) for PCR (Polymerase Chain Reaction), the biochip 100 is made of a material with small spontaneous fluorescence. It is desirable to be formed by. Examples of such a material having a small spontaneous fluorescence include polycarbonate and polypropylene. When the biochip 100 is used as a PCR reaction container, the biochip 100 is preferably made of a material that can withstand heating in PCR.

  Further, the biochip 100 may be made of carbon black, graphite, titanium black, aniline black, Ru, Mn, Ni, Cr, Fe, Co or Cu oxide, Si, Ti, Ta, Zr or Cr. Black substances such as carbides can be blended. By mixing such a black substance into the material of the biochip 100, the spontaneous fluorescence of the resin or the like can be further suppressed. In addition, when the biochip 100 is used from the outside of the biochip 100 for an application in which the inside of the internal cavity is observed (for example, real-time PCR), the material of the biochip 100 is made transparent as necessary. Can be. In addition, when the biochip 100 is used as a PCR reaction chip, the biochip 100 is preferably made of a material that has little nucleic acid or protein adsorption and does not inhibit an enzyme reaction such as polymerase.

  The first cavity 10 is a cavity formed inside the biochip 100. The shape of the first cavity 10 is not particularly limited. The shape of the first cavity 10 can be, for example, an elongated shape as shown in FIG. When the first cavity 10 has an elongated shape, for example, when the temperature of the biochip 100 is controlled so that regions having different temperatures are provided in the first cavity 10, the distance between the regions having different temperatures can be increased. It becomes easy to separate. Further, when the first cavity 10 has an elongated shape, the ratio of the surface area of the container to the volume of the container increases. For example, when the first cavity 10 is filled with a liquid such as oil. Therefore, the efficiency of heat conduction is improved, and the temperature control of the liquid can be facilitated. In the present embodiment, in the biochip 100, the shape of the first cavity 10 is elongated so that the direction of the straight line connecting the center portion of the first cavity 10 and the center portion of the second cavity 20 becomes the longitudinal direction. ing. Accordingly, when the biochip 100 is mounted on the rotating unit 210 such that the longitudinal direction thereof coincides with the radiation direction from the center of the thermal cycler 200, it is easy to provide regions of different temperatures in the first cavity 10. It becomes. In the biochip 100, the position where the first cavity 10 is provided is not limited as long as the second cavity 20 can be provided.

  The first cavity 10 can be filled with a liquid such as oil. A mechanism capable of injecting a liquid into the first cavity 10 may be provided at a portion of the biochip 100 that forms the first cavity 10. As such a mechanism, it can be comprised by the hole which connects the 1st cavity 10 and the exterior, and the member which plugs this, for example.

  One of the functions of the first cavity 10 is to become a reaction chamber for the liquid when the liquid is filled. For example, when the first cavity 10 is filled with oil and a PCR reaction solution, the first cavity 10 can be a space for reacting the PCR reaction solution. In particular, when the first cavity 10 is elongated, by moving the PCR reaction solution in the first cavity 10, it becomes easy to perform a thermal cycle on the PCR reaction solution.

  The second cavity 20 is a cavity formed inside the biochip 100. The second cavity 20 is provided adjacent to the first cavity 10. The shape of the second cavity 20 is not particularly limited, and may be a spherical shape, a cylindrical shape, or the like. In the illustrated example, the shape of the second cavity 20 is a rectangular parallelepiped shape.

  The second cavity 20 can be filled with a liquid such as oil. A mechanism capable of injecting a liquid into the second cavity 20 may be provided in a portion of the biochip 100 that forms the second cavity 20. As such a mechanism, it can be comprised by the hole which connects the 2nd cavity 20 and the exterior, and the member which plugs this, for example.

  One of the functions of the second cavity 20 is to collect the bubbles present in the first cavity 10 or the second cavity 20 in the second cavity 20 when filled with a liquid such as oil. Can be mentioned. For example, when oil and PCR reaction liquid droplets are filled in the first cavity 10 and the second cavity 20 and bubbles are mixed, the bubbles are collected in the second cavity 20. In addition, the collected bubbles can be prevented from returning to the first cavity 10. In other words, the liquid (oil) filled in the second cavity 20 moves to the first cavity 10 instead of the bubbles, so that bubbles that hinder the movement of the PCR reaction solution are removed from the first cavity 10. It can be removed and a good thermal cycle can be applied to the droplets of the PCR reaction solution.

  The opening / closing mechanism 30 can communicate or block the first cavity 10 and the second cavity 20. The opening / closing mechanism 30 operates based on gravity. Here, “operating based on gravity” includes, for example, that an opening / closing operation is caused directly by gravitational acceleration. Further, the buoyancy of bubbles in the liquid caused by the gravitational acceleration and the opening / closing mechanism 30 are It is an expression including the operation of opening and closing due to the buoyancy or mass of the constituent members. Since the direction in which the gravitational acceleration is applied to the biochip 100 can be changed depending on the posture of the biochip 100 (arrangement direction with respect to the direction of the gravitational acceleration), for example, the opening / closing mechanism can be changed by changing the posture of the biochip 100. 30 can be operated.

  In the example of the biochip 100 (FIG. 1), the opening / closing mechanism 30 includes a movable plug 32, and the movable plug 32 moves according to the direction in which gravity acts on the biochip 100. The adjacent portions of the cavity 10 and the second cavity 20 are open or closed.

  In the example of the biochip 100, the opening / closing mechanism 30 shows a mode in which the movable plug 32 can move along the direction of a straight line connecting the central portion of the first cavity 10 and the central portion of the second cavity 20. However, the direction of operation of the movable stopper 32 is not limited to this. For example, the movable plug 32 may operate along a direction intersecting with a direction of a straight line connecting the center portion of the first cavity 10 and the center portion of the second cavity 20. The moving direction and arrangement of the movable plug 32 can be selected as appropriate.

  In the biochip 100, the movable plug 32 is provided in the second cavity 20 together with the support portion 34 that supports the movable plug 32 (see FIG. 5). When the movable plug 32 moves to the first cavity 10 side, the portion connecting the first cavity 10 and the second cavity 20 can be closed. Thereby, the 1st cavity 10 and the 2nd cavity 20 can become an independent closed space, respectively. On the other hand, when the movable plug 32 moves to the second cavity 20 side, the portion connecting the first cavity 10 and the second cavity 20 can be opened. Thereby, the 1st cavity 10 and the 2nd cavity 20 can be connected, and can become a continuous closed space.

  In the biochip 100, the movable plug 32 has a material or structure that has buoyancy with respect to a liquid such as oil to be filled. Examples of such a movable plug 32 include those formed of resin foam and those having a hollow structure. Therefore, in the biochip 100, when the liquid is filled therein, the movable plug 32 operates by buoyancy based on gravity.

1.2. Thermal Cycler FIG. 2 is an exploded perspective view schematically showing the rotating unit 210 of the thermal cycler of the present embodiment. FIG. 3 is a perspective view schematically showing the rotating unit 210 of the thermal cycler of the present embodiment.

  When the biochip of this embodiment is used as a PCR reaction container, it can be used in a thermal cycler to perform a very good thermal cycle on the PCR reaction solution.

  The thermal cycler includes a rotating unit 210 that can change the posture of the biochip and supply heat to the first cavity 10. The thermal cycler described below is an example of a thermal cycler having such a mechanism. In this specification, such a thermal cycler is sometimes referred to as an “elevating type” thermal cycler.

  2 and 3, the thermal cycler has a cylindrical rotating part 210 having a central axis R, and the rotating part 210 is rotated around the central axis R by a drive mechanism (such as a motor) not shown. Has a structure to rotate. Moreover, as shown in FIG. 4, the rotation part 210 has the temperature control part 211 in the center axis | shaft R vicinity and the cylindrical periphery part, respectively. The number of the temperature control units 211 provided is not particularly limited. The temperature control part 211 can be appropriately formed in an annular shape. That is, a desired thermal cycle can be realized by adjusting the number of the temperature adjusting units 211, for example, a thermal cycle at a two-level temperature or a three-level temperature according to the number of the temperature adjusting units 211. Thermal cycle can be realized. The temperature adjusting unit 211 can be a heating element (such as a heater) or a heat absorbing element (such as a Peltier element). Moreover, the temperature set in each temperature adjustment unit 211 can also be set as appropriate.

  The rotating unit 210 has a mounting hole 220 into which a biochip can be loaded. In the illustrated example, a plurality of mounting holes 220 are formed on the cylindrical outer peripheral surface of the rotating unit 210. The number of mounting holes 220 formed is not limited.

  The biochip can be temperature-adjusted so as to have a different temperature for each part of the biochip by being loaded into the mounting hole 220 of the rotating part 210 of the thermal cycler. In addition, the biochip is loaded in the mounting hole 220 and the rotating portion 210 of the thermal cycler is rotated, whereby the posture of the biochip with respect to the direction of gravity acceleration can be changed. In this case, the rotation speed of the rotating unit 210 is preferably such that the centrifugal force applied to the biochip does not become too large, for example, 1 rpm or more and 10 rpm or less is preferable. However, when it is desired to apply centrifugal force to the biochip, the rotation speed of the rotating unit 210 is not limited to this.

  Then, when the biochip is filled with oil and PCR reaction liquid droplets, the thermal cycler is loaded into the rotating unit 210 and the rotating unit 210 is rotated. The droplet moves in the biochip based on gravity. In this case, in order to change the posture of the biochip with respect to the direction of gravity acceleration, the central axis R is installed so as not to be parallel to the direction of gravity acceleration.

1.3. Bubble Recovery Method FIG. 4 is a diagram schematically illustrating a state in which the biochip 100 of the present embodiment is loaded into the rotating unit 210 of the thermal cycler and rotated. In the illustrated example, the central axis R is orthogonal to the direction of gravity, and is drawn so that the upper side is a position with high potential energy in the figure. In addition, it is assumed that the rotation direction of the rotating unit 210 rotates clockwise around the central axis R. In the following description, when the position where the biochip 100 is present when the rotating unit 210 is rotating is described, the vertical upward direction in the figure is defined as an angle of 0 °, and 360 ° in one clockwise rotation from 0 °. It shall be expressed using angles. In the following description, the top and bottom are the same as the top and bottom directions in the figure.

  Hereinafter, the case where the biochip 100 is filled with oil and the PCR reaction solution w and the bubbles v exist in the first cavity 10 will be described. As the kind of oil, one having a specific gravity smaller than that of the PCR reaction solution w and incompatible with the PCR reaction solution w is used. The PCR reaction solution w includes, for example, a target nucleic acid (nucleic acid to be amplified), a primer (nucleic acid) for amplifying the target nucleic acid, an enzyme that performs a nucleic acid amplification reaction, and a fluorescent reagent for measuring the amount of amplification product (for example, SYBR GREEN) (Trademark)) and, if necessary, a liquid containing other nucleic acids. In the biochip 100, the liquid-liquid phase separation is performed in a form in which the PCR reaction liquid w becomes droplets in the oil, and further, the liquid (oil and PCR reaction liquid w) and the bubbles v are gas-liquid phase separated. It shall be.

  First, the state where the biochip 100 exists at the 0 ° position of the rotating unit 210 will be described. In FIG. 4, the PCR reaction solution is represented by the symbol w, and the bubbles are represented by the symbol v.

  When the biochip 100 is at the 0 ° position, both the movable plug 32 and the bubble v have buoyancy. Therefore, the movable plug 32 opens between the first cavity 10 and the second cavity 20, and is a space where both communicate. Therefore, the bubble v moves from the first cavity 10 to the second cavity 20 by buoyancy. Further, when the bubble v exists in the second cavity 20, it remains in the second cavity 20 as it is. On the other hand, at this time, the PCR reaction solution w moves to the end of the first cavity 10 opposite to the second cavity 20 because the specific gravity is greater than that of the oil. The movable stopper 32 does not operate until the rotation unit 210 rotates clockwise and the biochip 100 reaches the 90 ° position, and the bubble v is located at a position along the upper wall surface in the second cavity 20. The PCR reaction solution w exists at a position along the lower wall surface of the first cavity 10. Note that when the biochip 100 is in a position between 0 ° and 90 ° and a bubble cannot move from the first cavity 10 to the second cavity 20 (for example, at a position of 60 ° in FIG. 4). When air bubbles are generated in the first cavity 10, the rotating part 210 further rotates clockwise, and the biochip 100 comes to a position of 270 ° to 0 ° (360 °). Sometimes moved to the second cavity 20.

  When the position of the biochip 100 exceeds 90 ° due to the rotation of the rotating unit 210, the movable plug 32 receives buoyancy in the direction in which it is moved to the first cavity 10 side. The movable stopper 32 moves and closes between the first cavity 10 and the second cavity 20 until reaching the position of at least 180 °. The timing at which the movable stopper 32 moves may be between 90 ° and 180 °, and the position at which the movable stopper 32 is moved can be appropriately designed. Further, in this case, by adjusting the curvatures of the inner wall surfaces of the first cavity 10 and the second cavity 20, it is designed so that the bubbles v do not reach the communicating portion while the movable plug 32 is open. You can also

  When the biochip 100 reaches a position of 180 °, since the space between the first cavity 10 and the second cavity 20 is closed, the bubbles v in the second cavity 20 are buoyant and the bubbles v are caused by the buoyancy. The PCR reaction solution w in the first cavity 10 remains in the first cavity 10 without falling into the second cavity 20. .

  The movable plug 32 does not operate until the rotation unit 210 further rotates clockwise and the biochip 100 reaches the position of 180 ° to 270 °, and the bubbles v are generated on the upper wall surface in the second cavity 20. The PCR reaction solution is present at a position along the lower wall surface of the first cavity 10. In addition, when a bubble is generated in the first cavity 10 when the biochip 100 is at a position between 180 ° and 270 °, the rotation unit 210 further rotates clockwise when the bubble is generated in the first cavity 10. When the biochip 100 reaches a position of 270 ° to 0 ° (360 °), the biochip 100 is moved to the second cavity 20.

  Subsequently, when the position of the biochip 100 exceeds 270 °, the movable plug 32 receives buoyancy in the direction in which it is moved to the second cavity 20 side. Then, the movable stopper 32 moves and opens between the first cavity 10 and the second cavity 20 until reaching the position of at least 0 °. The timing at which the movable plug 32 moves may be between 270 ° and 0 ° (360 °), and the position at which the movable plug 32 is moved can be appropriately designed.

  As described above, in the biochip 100, each time the rotating unit 210 rotates and reaches the position of 0 °, the bubble v moves to the second cavity 20, and the bubble v then moves to the first cavity 10. It has never moved. That is, the biochip 100 can prevent the bubbles v existing in the biochip 100 from being collected in the second cavity 20 and returned to the first cavity 10.

  In the above, the biochip 100 is loaded in the rotating unit 210 of the thermal cycler, and the effect that the bubbles v in the first cavity 10 are removed by the rotation of the rotating unit 210 is exemplified. It will be easily understood that this is achieved not only by the change of the posture of the biochip 100 by the cycler but also if the posture of the biochip 100 with respect to the direction of gravitational acceleration can be changed by other methods.

  In the above example, in the biochip 100, the direction of the straight line connecting the central portion of the first cavity 10 and the central portion of the second cavity 20 is the longitudinal direction of the elongated shape of the first cavity 10. ing. The rotating unit 210 of the thermal cycler includes an annular temperature adjusting unit 211 and a central temperature adjusting unit 211 along the outer periphery. Therefore, the temperature at each position of the first cavity 10 of the biochip 100 can be set to be different. In the above example, the PCR reaction solution w is introduced into the biochip 100, and the PCR reaction solution w can also move within the first cavity 10. Therefore, if such an aspect is taken, a thermal cycle for the PCR reaction solution w can be realized. That is, according to the biochip 100, the PCR reaction solution w can be subjected to a thermal cycle while collecting the bubbles v. Therefore, the movement of the PCR reaction solution w in the first cavity 10 is prevented from being hindered by the bubbles v, and a more stable thermal cycle can be applied to the PCR reaction solution w.

1.4. Modifications of Biochip Although modifications are described below, the same components as those described in the above embodiment are denoted by the same reference numerals, and detailed description thereof is omitted. Moreover, in the structure described in the following modification, the structure shown with the same code | symbol as the above-mentioned embodiment can have the same effect | action function.

1.4.1. Deformation of First Cavity FIG. 5 is a partial cross-sectional perspective view schematically showing a biochip 110 in which the first cavity 10 is deformed. FIG. 6 is a schematic diagram of a cross section of the biochip 110. FIG. 7 is a schematic view of a cross section of the biochip 120 in which the first cavity 10 is deformed.

  The biochip of this embodiment can have a partition member 12 in the first cavity 10. As shown in FIGS. 5 and 6, the biochip 110 has a partition member 12 formed in the first cavity 10.

  The partition member 12 is formed so as to connect two spaced apart locations on the inner wall of the first cavity 10. That is, the partition member 12 is formed so as to connect a part of the inner wall of the first cavity 10 and another part spaced apart from the part of the inner wall. Therefore, the first cavity 10 can be a loop-shaped cavity by the partition member 12. That is, a path around the partition member 12 is formed in the first cavity 10 in which the partition member 12 is formed. A plurality of such circulating paths may be formed. That is, as shown in FIG. 7, the plurality of partition members 12 can form a cavity having a plurality of loop-shaped paths. The partition member 12 can be formed of the same material as the biochip 110.

  In the biochip 110 and the biochip 120, the partition member 12 is formed in the first cavity 10. Therefore, the oil, the PCR reaction solution w, and the bubbles v (gas phase) are present in the first cavity 10, and the 90 ° or 270 ° position of the rotating part 210 of the thermal cycler described above. When the rotating part 210 is loaded so that the positional relationship between the path of movement of the bubble v and the path of movement of the PCR reaction solution w in FIG. The path along which the PCR reaction solution w moves can be separated (see FIGS. 9 and 11). Thereby, when moving the bubble v and the PCR reaction solution w, the bubble v and the PCR reaction solution w can be moved without passing each other directly. Therefore, the movement of the PCR reaction solution w in the first cavity 10 is further reduced by the bubbles v, and a more stable thermal cycle can be applied to the PCR reaction solution w. Further, since the bubbles v and the PCR reaction solution w do not pass each other directly, both the path of movement of the bubbles v and the path of movement of the PCR reaction solution w can be narrowed. As a result, the movement (flow) of the oil accompanying the movement of the bubbles v or the PCR reaction solution w can be reduced, and the temperature distribution set in the first cavity 10 can be prevented from being disturbed. The accuracy of the cycle can be further improved.

  When the biochip has the partition member 12, the following modifications are possible. FIG. 8 is a schematic view of a cross section of the biochip 130 in which the first cavity 10 is deformed. The biochip 130 has the partition member 12 described above in the first cavity 10, and the crank portion 14 is formed in the formed loop path. In the illustrated example, the crank portion 14 is formed at two locations near the second cavity 20 side of the first cavity 10 and the center in the longitudinal direction of the first cavity 10. By forming the crank portion 14 in the first cavity 10, the following effects can be obtained.

  FIG. 9 is a diagram schematically illustrating a state in which the biochip 130 is loaded into the rotating unit 210 of the thermal cycler and rotates. As shown in FIG. 9, when the biochip 130 is loaded in the rotating unit 210 and the rotating unit 210 of the thermal cycler 200 is rotated, the PCR reaction solution w stays in the first cavity 10 and the position. You can control the time you stay. That is, by adjusting the shape of the crank portion 14 and the position where the crank portion 14 is provided, the PCR reaction solution w can stay at a position where the distance from the central axis R of the rotating portion 210 is different for a certain period of time. In the example shown in FIG. 9, while the biochip 130 rotates from the 330 ° position to the 60 ° position via 0 °, the position close to the central axis R (the second cavity of the first cavity 10). Stays at the crank portion 14 near the center of the first cavity 10 at a position of 90 ° to 210 °. Then, at a position from 210 ° to 300 °, it stays at a position far from the central axis R (the end of the first cavity 10 on the second cavity 20 side). Even in this case, when bubbles are generated in the first cavity 10, the bubbles are collected in the second cavity 20 by the opening / closing mechanism as in the above-described embodiment. In the example shown in the figure, a case where a bubble v is generated in the first cavity 10 at a position of 60 ° is shown, and the bubble v is collected in the second cavity 20 at a position near 0 ° (360 °). An example is shown.

  In this case, as shown in the figure, the temperature adjusting portion 211 of the thermal cycler 200 is located close to the central axis R (the end of the first cavity 10 opposite to the second cavity 20), the first cavity. 10 near the center and at a position far from the central axis R (the end of the first cavity 10 on the second cavity 20 side), while the rotating part 210 rotates once in the PCR reaction solution w. Three levels of temperature control can be performed with more accurate time distribution. In this case, the temperature set in each temperature adjustment unit 211 is arbitrary as long as PCR can be performed, and can be set to 94 ° C., 55 ° C., and 73 ° C., for example. If the arrangement of the crank portion 14 in the temperature adjusting unit 211 or the first cavity 10 is selected so that the temperature of the PCR reaction solution w is controlled in this order, the thermal denaturation, annealing, and extension reaction of the PCR reaction can be further performed. It can be suitably performed.

1.4.2. Modification of Opening / Closing Mechanism FIG. 10 is a cross-sectional view schematically showing a biochip 140 in which the opening / closing mechanism 30 is modified. FIG. 11 is a diagram schematically illustrating a state in which the biochip 140 is loaded in the rotating unit 210 of the thermal cycler and rotates.

  The opening / closing mechanism 30 included in the biochip 140 according to the modification is a valve 36. The valve 36 can be opened from the first cavity 10 side toward the second cavity 20 side, and is provided to be closed from the second cavity 20 side toward the first cavity 10 side. The valve 36 operates based on gravity. Specifically, the valve 36 operates by the buoyancy of bubbles when bubbles are present in the biochip 140. A partition member 14 is formed in the first cavity 10 of the biochip 140. The valve 36 is formed of a material having moderate elasticity.

  Hereinafter, a case where the biochip 140 is filled with oil and the PCR reaction solution w and the bubbles v are present in the first cavity 10 will be described. The oil, the PCR reaction solution w, and the bubbles v are the same as described in “1.3. Method for recovering bubbles”.

  First, the state where the biochip 140 exists at the 0 ° position of the rotating unit 210 will be described with reference to FIG. Also in FIG. 12, the PCR reaction solution is represented by the symbol w, and the bubbles are represented by the symbol v.

  When the biochip 140 is in the 0 ° position, the valve 36 is pushed up and opened by the buoyancy of the bubble v, the first cavity 10 and the second cavity 20 communicate with each other, and the bubble v is buoyant. Move from the first cavity 10 to the second cavity 20. Further, when the bubble v exists in the second cavity 20, it remains in the second cavity 20 as it is. On the other hand, at this time, the PCR reaction solution w moves to the end of the first cavity 10 opposite to the second cavity 20 because the specific gravity is greater than that of the oil. Then, when the rotating unit 210 rotates clockwise or the bubble v moves to the second cavity 20 and no longer exists in the first cavity 10, the valve 36 is closed (30 ° in FIG. 11). And 60 ° position). At this time, the bubbles v collected at the 0 ° position are present along the upper wall surface in the second cavity 20, and the PCR reaction solution is present along the lower wall surface of the first cavity 10. It will be. In the example shown in the figure, when the biochip 140 is at a position other than near 0 ° and the air bubbles cannot move from the first cavity 10 to the second cavity 20, the biochip 140 enters the first cavity 10. When bubbles are generated, the bubbles are moved to the second cavity 20 when the rotating unit 210 further rotates clockwise and the biochip 100 comes to a position near 0 ° (360 °). . In the example shown in the figure, a case where a bubble v is generated in the first cavity 10 at a position of 60 ° is shown, and the bubble v is collected in the second cavity 20 at a position near 0 ° (360 °). An example is shown.

  Further, since the partition member 14 is provided in the first cavity 10 of the biochip 140, when both the bubbles v and the PCR reaction solution w exist in the first cavity 10, a loop shape is formed. It moves through paths opposite to each other in the flow path. As a result, the movement of the PCR reaction solution w in the first cavity 10 is further prevented from being hindered by the bubbles v, and a more stable thermal cycle can be applied to the PCR reaction solution w. The same as described in “1.4.1. Deformation of first cavity”.

  When the rotating unit 210 further rotates and the position of the biochip 100 reaches 180 °, the PCR reaction solution w receives gravitational acceleration in a direction to open the valve 36 toward the second cavity 20. However, since the specific gravity difference between the oil and the PCR reaction solution w is smaller than the specific gravity difference between the oil and the bubble v, at this position, the valve 36 is not opened, and the PCR reaction solution w is in the first cavity 10. stay. The force required to open the valve 36 can be designed by appropriately selecting the type of oil and PCR reaction solution w, the elasticity of the material of the valve 36, the size of the valve 36, and the like.

  When the rotating unit 210 further rotates clockwise, the biochip 100 reaches the position of 0 ° (360 °) again, and when bubbles are generated in one rotation until then, the valve 36 operates. Then, new bubbles are collected in the second cavity 20.

  As described above, in the biochip 140, every time the rotating unit 210 rotates and reaches the position of 0 °, the bubble v moves to the second cavity 20, and the moved bubble v then moves to the first cavity 10. It has never been moved to. That is, the biochip 140 can prevent the bubbles v existing in the biochip 140 from being collected in the second cavity 20 and returned to the first cavity 10.

  In the biochip of the present embodiment, when the elevating thermal cycler 200 is used, even when bubbles are present in the biochip 10, the opening / closing mechanism 30 operates based on gravity, whereby the second cavity 20. Bubbles can be collected inside the interior. Therefore, it is possible to prevent bubbles from being present in the first cavity 10. Thereby, for example, when oil and a PCR reaction solution are filled, a stable thermal cycle can be applied to the PCR reaction solution in the first cavity 10.

2. Second Embodiment A second embodiment will be described below. The same components as those described in the first embodiment are denoted by the same reference numerals, and detailed description thereof is omitted. Moreover, in the structure described in 2nd Embodiment, the structure shown with the same code | symbol as 1st Embodiment can have the same effect | action function.

  FIG. 12 is a schematic view of a cross section of the biochip 150 of the present embodiment. 13 and 14 are diagrams schematically showing the main part of the biochip. FIGS. 15, 16, and 17 are diagrams schematically illustrating how the biochip 150 is loaded in the rotating unit 210 of the thermal cycler and rotates.

  The biochip 150 of the present embodiment is a biochip used for a lift-type thermal cycler, and has a first cavity 10, a second cavity 20, a first barrier 41, and a second barrier 42. In addition, a communication path 40 is formed between the first cavity 10 and the second cavity 20.

  The first cavity 10 and the second cavity 20 are the same as described in “1. First embodiment”. The communication path 40 is formed between the first cavity 10 and the second cavity 20. The communication path 40 allows the first cavity 10 and the second cavity 20 to communicate with each other. The shape of the communication path 40 is not particularly limited.

  The first barrier 41 is a wall that is provided between the first cavity 10 and the communication path 40 and that protrudes from an inner wall surface that defines the first cavity 10 and the communication path 40. The first barrier 41 is formed so as to maintain the communication between the first cavity 10 and the communication path 40 without being blocked. The first barrier 41 can be formed of the same material as the biochip 150.

  The second barrier 42 is provided between the second cavity 20 and the communication path 40, and is opposite to the direction in which the first barrier 41 protrudes from the inner wall surface that defines the second cavity 20 and the communication path 40. It is a wall formed to project. The second barrier 42 is formed so as to maintain the communication between the second cavity 20 and the communication path 40 without being blocked. The second barrier 42 can be formed of the same material as the biochip 150.

  When the first barrier 41 and the second barrier 42 are viewed from the side of the first cavity 10 through the communication path 40 through the direction perpendicular to the first barrier 41 and the second barrier 42, The two cavities 20 are provided so as to be in a positional relationship shielded by the first barrier 41 and the second barrier 42. In other words, the first cavity 10 and the second cavity 20 cannot be seen through the communication passage 40, and the first barrier 41 and the second barrier 42 are like partitions that protrude in opposite directions. Yes.

  In order to explain the operation of the first barrier 41 and the second barrier 42 in more detail, FIG. 13 and FIG. 14 are referred to. FIGS. 13 and 14 are views showing a state in which the main part of the biochip (near the communication path 40, the first barrier 41, and the second barrier 42) rotates. The first cavity 10 and the second cavity 20 are shown in FIG. It is the figure drawn simplified. 13 and 14 also, the biochip is filled with oil and PCR reaction solution w. It is assumed that bubbles v1 and v2 exist in the biochip. The oil, the PCR reaction solution w, and the bubbles v1 and v2 are the same as described in “1.3. Method for recovering bubbles”.

  First, based on FIG. 13, the movement of the bubble v1, the bubble v2, and the PCR reaction solution w will be described. At the 0 ° position, as shown in the figure, the bubble v1 is present in the first cavity 10, and the bubble v2 and the PCR reaction solution w are present in the communication path 40. From this state, when rotating clockwise from the 0 ° position, the bubbles and the PCR reaction solution move in the biochip, and as shown in (b) at the 45 ° position, the bubble v1 and the PCR reaction solution are moved. w exists in the 1st cavity 10, and the bubble v2 comes to exist in the communicating path 40. FIG. Thereafter, the bubbles and the PCR reaction solution do not move to the 90 ° position, and move after 90 °.

  FIG. 13D shows a state immediately after the biochip passes the 90 ° position. At this time, the bubble v2 and the PCR reaction solution w move along the inner wall of the second cavity 20 and the inner wall of the first cavity 10, respectively, but the bubble v1 moves along the first barrier 41, When the first barrier 41 is interrupted, it floats on the wall surface of the second barrier 42 on the side of the communication path 40 by buoyancy, and then moves along the second barrier 42 (shown by a broken line in FIG. 13D). . Then, as shown by (e) at a position of 135 °, the bubble v1 is present in the communication path 40, the PCR reaction solution w is present in the first cavity 10, and the bubble v2 is present in the second cavity. 20 will be present. Thereafter, when the biochip rotates to a position of 270 °, as shown in FIG. 13 (h), the bubble v1 and the PCR reaction solution w exist in the first cavity 10, and the bubble v2 is in the second cavity. 20 will be present.

  (I) of FIG. 13 shows a state immediately after the biochip passes the position of 270 °. At this time, the bubble v <b> 1 moves along the inner wall of the first cavity 10. At the same time, the bubble v2 moves along the second barrier 42, and floats on the wall surface of the first barrier 41 on the side of the communication path 40 by buoyancy where the second barrier 42 is interrupted, and then along the first barrier 41. Move. At the same time, the PCR reaction solution w moves along the first barrier 41 and falls to the wall surface of the second barrier 42 on the side of the communication path 40 due to gravity where the first barrier 41 is interrupted, and then the second barrier 42. Move along. Then, as indicated by (j) at a position of 315 °, the bubble v1 exists in the first cavity 10, and the bubble v2 and the PCR reaction solution w exist in the communication path 40, This is the state at the 0 ° position.

  As described above, it will be understood that both the bubble and the PCR reaction solution move within the cavity in which they originally existed. Next, a method for moving only the bubble v2 to the second cavity 20 will be described with reference to FIG.

  FIG. 14 (e) corresponds to FIG. 13 (e) and shows a state at a position between 90 ° and 180 ° by the clockwise rotation in FIG. Here, the state in which the biochip is rotated in the reverse direction (counterclockwise) and is rotated to an angle smaller than 90 ° across the 90 ° position is (k) in FIG. At this position, the bubble v2 and the PCR reaction solution w move along the inner wall of the second cavity 20 and the inner wall of the first cavity 10, respectively, while the bubble v1 moves along the second barrier 42. When the second barrier 42 is interrupted, it floats toward the inner wall of the second cavity 20 by buoyancy. As a result, the bubbles v1 and the bubbles v2 are present in the second cavity 20, and the PCR reaction solution w is present in the first cavity 10. When the rotation direction of the biochip is reversed from clockwise to counterclockwise and the phenomenon occurs, the rotation direction is reversed again and the rotation direction of the biochip is rotated clockwise. If it does so, it will be in the state as shown to (m) of FIG. 14 in the position between 90 degrees and the position to 180 degrees. After that, both the bubbles v1 and the bubbles v2 move in the second cavity 20 through the same path as the path of the bubbles v2 described above, and do not return to the first cavity 10. Will move.

  In this way, the bubbles v1 in the first cavity 10 can be collected in the second cavity 20 by reversing the rotation direction at a specific position. Then, the recovered bubbles can be retained in the second cavity 20 without returning from the second cavity 20 to the first cavity 10. In addition, in the case where a plurality of biochips can be mounted on the rotating unit 210 of one thermal cycler, and when two biochips are provided symmetrically with respect to the central axis R, with respect to one biochip, If the above reversal operation across the 90 ° position is performed in order to collect the bubbles, the reverse operation across the 270 ° position is performed on the other biochip. However, in this case, since the specific gravity difference between the oil and the PCR reaction liquid is smaller than the specific gravity difference between the oil and the bubbles, the movement speed of the PCR reaction liquid is smaller than the movement speed of the bubbles. The PCR reaction solution can be prevented from moving to the second cavity 20.

  Here, the conditions for allowing the bubbles and the PCR reaction solution to move in the biochip as described above appear in (d) and (i) of FIG. In the movement of the bubbles and the PCR reaction solution in (d) and (i) of FIG. 13, it can be seen that after one barrier is interrupted, the other barrier only needs to be moved. That is, when the first barrier 41 and the second barrier 42 are viewed from the first cavity 10 side through the communication path 40 and in a direction perpendicular to the first barrier 41 and the second barrier 42, It is a condition that the second cavity 20 is provided so as to be in a positional relationship shielded by the first barrier 41 and the second barrier 42. On the other hand, the second barrier 42 and the first barrier 41 are seen from the direction perpendicular to the second barrier 42 and the first barrier 41 through the communication path 40 from the second cavity 20 side. Sometimes, the first cavity 10 may be provided so as to be in a positional relationship shielded by the second barrier 42 and the first barrier 41.

  Since the structure of the main part of the biochip has such a structure, the air bubbles v1 in the first cavity 10 are removed from the second cavity by the rotation method including the reversing operation of the biochip without requiring an opening / closing mechanism. It can be collected in the tee 20. Then, the recovered bubbles can be retained in the second cavity 20 without returning from the second cavity 20 to the first cavity 10.

  FIG. 15 is a diagram schematically illustrating a state in which the biochip 150 described above is loaded in the rotating unit 210 of the thermal cycler and rotates. As shown in FIG. 15, since the biochip 150 has the structure of the main part described above, the bubbles recovered in the second cavity 20 are secondly rotated by a clockwise rotation without requiring an opening / closing mechanism or the like. The cavity 20 can be retained in the second cavity 20 without returning to the first cavity 10. At the same time, a desired thermal cycle can be applied to the PCR reaction solution w in the first cavity 10 by clockwise rotation.

  FIG. 16 shows a state in which the rotating unit 210 is reversed across the 90 ° position. As shown in FIG. 16, the biochip 150 rotates counterclockwise across a 90 ° position, thereby removing bubbles in the first cavity 10 without requiring an opening / closing mechanism or the like. Can be recovered within. Then, as shown in FIG. 17, when the rotation direction is reversed again and the clockwise rotation is performed, the recovered air bubbles are not returned from the second cavity 20 to the first cavity 10, and are returned to the second cavity. 20 can be retained.

  When the biochip 150 is rotated by the thermal cycler 200 capable of loading a plurality of biochips 150, when bubbles are generated in the first cavities 10 of any of the biochips 150 during the clockwise rotation. In other words, when the biochip 150 in which the bubbles are generated reaches a position near 90 °, the bubbles can be collected by performing a reversal operation across the 90 ° position.

  In the biochip 150 of the present embodiment, when the elevating thermal cycler 200 is used, even if bubbles exist in the biochip 150, the configuration of the first barrier 41 and the second barrier 42 and the reversal operation in the rotation direction are performed. By the action of the above, it is possible to collect the bubbles in the second cavity 20 and prevent the bubbles from returning into the first cavity 10. Therefore, it is possible to prevent bubbles from being present in the first cavity 10. Thereby, for example, when oil and a PCR reaction solution are filled, a stable thermal cycle can be applied to the PCR reaction solution in the first cavity 10.

  The embodiments described above and the modified embodiments can be arbitrarily combined with a plurality of forms. Thereby, combined embodiment can have an effect which each embodiment has, or a synergistic effect.

  The present invention is not limited to the above-described embodiments, and various modifications can be made. For example, the present invention includes substantially the same configuration (for example, a configuration having the same function, method, and result, or a configuration having the same purpose and effect) as the configuration described in the embodiment. In addition, the invention includes a configuration in which a non-essential part of the configuration described in the embodiment is replaced. In addition, the present invention includes a configuration that achieves the same effect as the configuration described in the embodiment or a configuration that can achieve the same object. In addition, the invention includes a configuration in which a known technique is added to the configuration described in the embodiment.

DESCRIPTION OF SYMBOLS 10 ... 1st cavity, 12 ... Partition member, 14 ... Crank part, 20 ... 2nd cavity, 30 ... Opening / closing mechanism, 32 ... Movable stopper, 34 ... Support part, 36 ... Valve, 40 ... Communication path, 41 ... 1st barrier, 42 ... 2nd barrier, 100, 110, 120, 130, 140, 150 ... biochip, 200 ... thermal cycler, 210 ... rotating part, 211 ... temperature control part, 220 ... mounting hole, R ... central axis

Claims (6)

A first cavity filled with oil;
A second cavity adjacent to the first cavity and filled with the oil;
An opening / closing mechanism that operates based on gravity, and that connects or blocks the first cavity and the second cavity;
Having a biochip. In claim 1,
The open / close mechanism is a biochip having a movable stopper that opens or closes the space between the first cavity and the second cavity. In claim 1,
The biochip includes a valve that is opened from the first cavity side toward the second cavity side and is closed from the second cavity side toward the first cavity side. In any one of Claims 1 to 3,
The first cavity is a biochip having a shape whose longitudinal direction is a direction connecting the first cavity and the second cavity. In any one of Claims 1 thru | or 4,
A partition member is formed in the first cavity,
A biochip having a path through which the oil can circulate around the partition member in the first cavity. A first cavity filled with oil;
A second cavity that communicates with the first cavity via a communication path and is filled with the oil;
A first barrier formed between the first cavity and the communication path and protruding from an inner wall surface defining the first cavity and the communication path;
A direction parallel to the first barrier is formed between the second cavity and the communication path, and is opposite to a direction in which the first barrier protrudes from an inner wall surface defining the second cavity and the communication path. A second barrier formed protruding in the direction of
Have
The first cavity, the communication path, and the second cavity are continuous closed spaces,
When the second cavity is viewed from a direction perpendicular to the first barrier and the second barrier through the communication path from the first cavity side, the second cavity is the first barrier and A biochip shielded by the second barrier.

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