JP6012518B2 - Temperature control mechanism for biochemical cartridge, temperature control block, and biochemical processing equipment - Google Patents

Description

  The present invention relates to a temperature control mechanism for a biochemical cartridge, a temperature control block, and a biochemical processing apparatus suitable for amplifying a DNA (deoxyribonucleic acid) fragment by PCR (Polymerase Chain Reaction).

  2. Description of the Related Art In recent years, research has been conducted to react a sample with a reagent in a microspace of a microfluidic device to perform a chemical reaction (extraction, purification, amplification, etc.) and analysis. Microfluidic devices can be applied to a wide range of applications such as gene analysis. In addition, the use of microfluidic devices (1) requires less sample and reagent consumption compared to normal equipment, (2) is easier to carry than when various reagents are set, (3 ) It has the advantage of being disposable.

  Genetic analysis requires biochemical treatments and reactions such as extracting and amplifying nucleic acids such as DNA and RNA (ribonucleic acid) from samples obtained from living organisms. In these treatments and reactions, temperature may be an important factor, and it is necessary to heat or cool the sample as necessary.

  PCR is an example of a biochemical process or reaction where temperature is an important factor. PCR is an enzyme chain reaction and is an important molecular biology technique for amplifying DNA. In PCR, DNA is amplified by applying a three-stage or two-stage temperature cycle to an appropriately prepared reaction solution. The temperature cycle is an operation of repeating different temperature states called heat denaturation, annealing, and elongation, and temperatures around 95 ° C., 60 ° C., and 72 ° C. are used, respectively. The time for maintaining each temperature is several seconds to several minutes. The number of cycles is generally around 30.

  The time required for normal PCR is about 90 minutes. In order to shorten the time required for PCR, FastPCR has been devised in which the maintenance time at each temperature is shortened and the time required for PCR is shortened to 30 to 45 minutes. In order to further shorten the maintenance time, it is necessary to quickly change the reaction solution to the target temperature. On the other hand, when PCR is performed for the purpose of gene analysis or the like, it is necessary to prevent contamination of DNA not to be analyzed. This is because, if DNA not to be analyzed is mixed from outside during PCR, the mixed DNA is also amplified. For this reason, the microfluidic device employs a structure in which reagent adjustment, PCR and the like can be operated in a closed system shut off from the outside, reducing the possibility of contamination. For heating and cooling of the sample fluid on the microfluidic device, for example, a temperature control mechanism described in Patent Document 1 is used.

JP 2011-30522 A

  The temperature control mechanism disclosed in Patent Document 1 forms a concave portion for adjusting the sample temperature in a semicircular cross section, and adjusts the sample temperature through contact between the membrane layer and the concave portion that are deformed by its own weight as the sample is injected. Realize. However, in this method, the membrane layer and the entire inner peripheral surface of the recessed portion are not necessarily in close contact with each other, and there is a possibility that heat exchange with the sample fluid may be hindered. Therefore, the present invention provides a temperature control mechanism for a biochemical cartridge capable of efficiently exchanging heat with a sample fluid.

  The temperature control mechanism for a biochemical cartridge according to the present invention assumes mounting of a biochemical cartridge in which a membrane, which is an elastic body that can be deformed by air pressure, is attached to the surface. In order to solve the above problems, the present invention provides a reaction structure forming concave structure that faces an opening end of a flow path formed on the biochemical cartridge side across a membrane when the biochemical cartridge is mounted, A temperature control block having at least one air suction opening provided on the inner peripheral surface of the concave structure and a pipe connected to the opening, and a pump for sucking air from the concave structure through the pipe And have.

  According to the present invention, the reaction part can be formed by deforming the membrane so as to be in close contact with the concave structure on the temperature control block side, and the temperature of the reaction liquid held in the membrane can be adjusted efficiently. Problems, configurations, and effects other than those described above will become apparent from the following description of embodiments.

The figure which shows the structural example of the pre-processing integrated capillary electrophoresis apparatus which concerns on an Example. The figure which shows the structural example of the pre-processing mechanism which concerns on an Example. The figure explaining the structure before and behind the reaction part which concerns on an Example. The figure explaining schematic planar structure of the temperature control mechanism which concerns on an Example. The enlarged view explaining the structural example of the reaction well which concerns on an Example, and its periphery. The A-A 'sectional view of the temperature control mechanism concerning an example. B-B 'sectional drawing of the temperature control mechanism which concerns on an Example. The flowchart which shows an example of the liquid feeding method to the temperature control part which concerns on an Example. The figure explaining the liquid feeding to the temperature control part which concerns on an Example. The flowchart which shows an example of the liquid feeding method to the temperature control part which concerns on an Example. The flowchart which shows an example of the liquid feeding method to the temperature control part which concerns on an Example. The figure explaining the liquid feeding from the temperature control part which concerns on an Example. FIG. 6 is a diagram illustrating a configuration of a temperature control mechanism according to a second embodiment.

  Hereinafter, embodiments of the present invention will be described with reference to the drawings. The embodiment of the present invention is not limited to the examples described later, and various modifications are possible within the scope of the technical idea.

[Example 1]
(Overall configuration of the device)
In this embodiment, a pretreatment integrated capillary electrophoresis apparatus used for DNA analysis will be described as an example of a biochemical treatment apparatus. FIG. 1 shows a configuration example of a pretreatment integrated capillary electrophoresis apparatus 1. The pretreatment integrated capillary electrophoresis apparatus 1 includes a pretreatment unit 2 and an analysis unit 3. The pretreatment unit 2 performs pretreatment for extracting and amplifying DNA from the reaction solution, and the analysis unit 3 analyzes the DNA treated by the pretreatment unit 2 by capillary electrophoresis. The detailed configuration of the preprocessing unit 2 will be described later. The analysis unit 3 includes a capillary 5, a detection unit 6, an oven 8, and a high voltage power source 9.

  The pretreatment integrated capillary electrophoresis apparatus 1 drives the pretreatment unit 2 in the horizontal direction through the autosampler 4 after completion of the pretreatment, and connects one end of the flow path of the pretreatment unit 2 to one end of the capillary 5 of the analysis unit 3. Connect with. The capillary 5 is filled with a polymer and is kept at a constant temperature by an oven 8. After the connection of the capillary 5, the analysis unit 3 applies a high voltage to both ends of the capillary 5 by the high voltage power source 9, electrophoreses the DNA sample introduced into the capillary, and the detection unit 6 performs fluorescence analysis.

(Configuration of pre-processing unit)
FIG. 2 shows a configuration example of the preprocessing unit 2. The pretreatment unit 2 includes a liquid feeding mechanism 12 and a temperature adjustment mechanism 13. 2 shows a state where a cartridge 11 as a microfluidic device is mounted. The flat cartridge 11 is formed with a channel sealed from the outside by a membrane, and biochemical processing from DNA extraction to amplification is executed in the channel. The liquid feeding mechanism 12 is a mechanism (pump 7 in FIG. 1) that feeds a fluid such as a sample or a reagent through a support surface (mounting surface) that supports the cartridge 11 and a flow path formed in the cartridge 11. Including). The temperature adjustment mechanism 13 is composed of, for example, a Peltier element, and controls the fluid in the cartridge 11 to a temperature suitable for PCR.

  FIG. 3 shows a cross-sectional structure of the cartridge 11 and the pretreatment unit 2 in the mounted state. The cartridge 11 includes a well 24a that encloses a reagent used for each treatment, a narrow channel 23a for sending the reagent from the well 24a to the reaction unit that performs PCR, and a reaction solution from the reaction unit to the well 24b. Narrow flow path 23b for this purpose is formed.

  A membrane 21, which is an elastic body that can be deformed by air pressure, is attached to the lower surface of the cartridge 11 (the surface to be attached to the pretreatment unit 2). The membrane 21 includes at least the opening of the well 24a and one opening of the narrow channel 23a, the other opening of the narrow channel 23a and one opening of the narrow channel 23b, and the other opening of the narrow channel 23b. What is necessary is just to affix so that the opening of well 24b may be connected and a flow path may be formed. As will be described later, the membrane 21 is formed in the liquid supply passage grooves 22a and 22b formed on the support surface (mounting surface) of the liquid supply mechanism 12 and the groove (reaction well 34 described later) that defines the outer shape of the reaction portion. By deforming along, a flow path for liquid feeding and a reaction part are formed. That is, the reaction solution in the cartridge 11 is sent from the well 24 a to the well 24 b through a flow path formed by deformation of the membrane 21. That is, if the cartridge 11 is used, the reaction solution can be fed and reacted in a state where contamination with the outside air is eliminated.

The cartridge 11 is made of, for example, a material such as polycarbonate resin (PC), polypropylene resin (PP), or cycloolefin resin (COP) that has resistance to both heat and chemicals. The membrane 21 is made of a rubber material such as silicon rubber, ethylene propylene diene rubber (EPDM) or butyl rubber, or a material such as an elastomer material. Furthermore, in order to perform stable analysis while preventing evaporation as much as possible, the membrane 21 is made of a material having low water vapor permeability and gas permeability. For example, a membrane having a gas permeability of 20 cc · cm / cm ² · sec · atm or less is used.

  On the other hand, the liquid feeding mechanism 12 includes liquid feeding flow path grooves 22a and 22b having a groove structure adapted to the structure of the cartridge 11, and a suction port 25 and a pressure port 26 that deform the membrane 21 into a flow path shape by air pressure. Is provided. The suction port 25 and the pressurization port 26 are connected to a pump 7 (FIG. 1) that controls the air pressure via a connection portion (not shown) and a piping tube. A switching valve such as a three-way valve is disposed in the connection portion, and the application timing of air to the suction port 25 and the pressure port 26 is controlled through opening and closing of the switching valve. The pump 7 used in the present embodiment is, for example, a diaphragm type or rotary type air pump, and can generate −1 MPa to 1 MPa.

  The liquid feeding method will be described with reference to FIG. When the air in the space surrounded by the liquid feeding channel groove 22a and the membrane 21 is sucked from the suction port 25, the membrane 21 is deformed along the surface of the liquid feeding channel groove 22a and is connected to the well 24a. A path is formed. The reaction solution is introduced from the well 24a into the liquid supply channel 22a thus formed. On the other hand, when air is introduced between the membrane 21 and the liquid-feeding channel groove 22a from the pressurizing port 26, the membrane 21 is pushed up and returns to a state of being in close contact with the lower surface of the cartridge 11. As a result, the reaction liquid introduced into the liquid sending channel groove 22a is sent to the reaction section through the narrow channel 23a.

  A temperature control mechanism 13 is disposed at a position where the reaction part is formed. The temperature control mechanism 13 includes a temperature control block 32, a heating / cooling device 51, and a heat dissipation block 52. The temperature control mechanism 13 is disposed between the two liquid feeding mechanisms 12 and radiates heat through a heat sink 53 (FIG. 6) attached to the heat dissipation block 52.

(Configuration of temperature control block)
FIG. 4 is a perspective view showing the structure of the temperature control block 32 as viewed from the mounting surface side of the cartridge 11. As shown in FIG. 4, the temperature control block 32 includes a reaction well 34, a heat insulating material 35, a pressure suction path 36, a pressure suction joint 31, and a temperature sensor 33. In the case of FIG. 4, eight grooves (reaction wells 34) that define the outer shape of the reaction part are formed on the surface of the temperature control block 32. The number of reaction wells 34 is determined according to the number of cartridges 11 to be mounted. In the temperature control block 32 according to the present embodiment, the heating process and the cooling process for the eight cartridges 11 can be executed simultaneously in parallel.

  In this embodiment, the reaction well 34 is formed in a hexagonal shape as shown in FIG. However, the shape of the reaction well 34 is not limited to the hexagonal shape, and may be other shapes (for example, a rectangle, a circle, and an ellipse). In the case of this embodiment, a total of 16 pressure suction ports 41a and 41b for controlling air pressure in the reaction well 34 are arranged at two apex positions of the six apex angles. The pressure suction ports 41a and 41b are arranged in the vicinity of the openings of the narrow channels 23a and 23b as shown in FIG.

  In the case of this embodiment, the membrane 21 at the corresponding position can be deformed along the surface shape of the reaction well 34 by sucking the air in the reaction well 34 through the pressure suction ports 41a and 41b. By this deformation, a reaction part is formed.

  In the case of FIG. 4, one pressure suction path 36 formed inside the heat dissipation block 52 is connected to the four pressure suction ports 41 a and 41 b located on the same vertex side. A pressure suction joint 31 is connected to each of the four pressure suction ports 41a and 41b. A pump 7 (FIG. 1) is connected to the pressure suction joint 31 through a valve such as a two-way valve or a three-way valve. By appropriately controlling these valves, the reaction well 34 can be pressurized or depressurized (suctioned) while using the same pressure suction port 41. The operation of the pump 7 is controlled by a control unit (not shown). In the case of the present embodiment, a resin adapter (not shown) is connected between the heat radiation block 32 and the pressure suction joint 31. This is to reduce heat radiation from the heat radiation block 32 to the pressure suction joint 31. A heat insulating material 35 is disposed around the outer periphery of the eight reaction wells 34. This is to keep the temperature of the eight reaction wells 34 uniform.

  6 and 7 show the AA cross section and the BB cross section of the temperature control block 32 shown in FIG. As shown in FIG. 6, the temperature sensor 33 is disposed below the reaction well 34 for measuring the reaction temperature of the eight reaction wells 34 arranged in a straight line. In the present embodiment, three temperature sensors 33 are arranged. However, the number of temperature sensors 33 may be two or less or four or more. In the case of the present embodiment, the number of temperature sensors 33 is provided according to the number of heating / cooling devices 51. This is for feedback control of each heating / cooling device 51 according to the measured temperature.

  As shown in FIG. 6, the heat insulating material 35 is also attached to the side surface portion of the temperature control block 32. By sticking the heat insulating material 35 to the side surface portion of the temperature control block 32, heat radiation from the side surface of the temperature control block 32 can be prevented.

  As shown in FIGS. 6 and 7, the heat dissipation block 52 is arranged so that the wind from the air-cooling fan 54 arranged at the lower position thereof is not directly hit. Incidentally, the wind of the air cooling fan 54 is set so as to directly hit the heat sink 53 attached to the lower surface of the heat dissipation block 52.

  Here, the cross-sectional structure of the reaction well 34 will be described. As shown in FIGS. 6 and 7, the cross-sectional shape of the reaction well 34 is rectangular, and has a structure in which the height of the groove (thickness in the direction of gravity) is smaller than the length in the liquid feeding direction. Yes. By reducing the height of the groove, the thermal resistance of the reaction solution is reduced, and a temperature difference is hardly generated between the upper surface side and the lower surface side of the reaction solution.

  In the case of Patent Document 1 described above, the concave portion corresponding to the reaction well 34 is formed in a semicircular cross section, the groove becomes deeper in proportion to the length in the liquid feeding direction, and the upper surface of the concave portion. A temperature difference is likely to occur between the reaction solution on the side and the reaction solution on the bottom surface side.

  As described above, the reaction well 34 according to the present embodiment can be made much thinner than the conventional structure, so that more accurate temperature adjustment is possible. In the case of the present embodiment, the reaction well 34 can be thinned, so that the temperature control block 32 can be thinned.

  Incidentally, the temperature control block 32 is made of a material having a certain level of rigidity, good thermal conductivity, and low specific heat (for example, aluminum alloy, copper, brass, etc.). Of course, in order to change the temperature of the temperature control block 32 quickly, it is desirable to keep the heat capacity of the temperature control block 32 as small as possible. For this reason, in the temperature control block 32 of the present embodiment, a structure in which a portion other than the portion into which the reaction well 34 and the temperature sensor 33 are inserted is adopted as necessary to reduce the heat capacity of the block itself.

  A known device is used as the heating / cooling device 51 of the temperature control mechanism 13. In this embodiment, a Peltier element that can be heated and cooled is used. One or a plurality of heating / cooling devices 51 are arranged in accordance with the size and shape of the temperature control block 32. However, depending on the application, a heating source and a cooling source may be arranged, or only one of them may be arranged.

  One or more temperature sensors 33 provided in the temperature control block 32 are provided depending on the number of heating / cooling devices 51 provided in the temperature control mechanism 13 and the size and shape of the temperature control block 32. In this embodiment, three temperature sensors 33 and a heating / cooling device 51 are provided. A platinum electrode, a thermistor, or a resistance temperature detector can be used for the temperature sensor 33, and a Peltier element can be used for the heating / cooling device 51.

  As shown in FIG. 7, the temperature sensor 33 and the heating / cooling device 51 are connected to a temperature control board 61. Detection signals from the temperature sensors 33 are input to the temperature control board 61. The temperature control board 61 transmits a control signal to each heating / cooling device 51 based on the calculation result, and controls its temperature. The temperature control board 61 includes elements necessary for temperature control (for example, a memory, a CPU, a circuit necessary for on / off control of the heating / cooling device 51, and the like).

  At the time of heating or cooling, the temperature control board 61 feedback-controls the heating / cooling device 51 based on all the measured values of the corresponding pair or provided temperature sensors 33. In the present embodiment, the temperature of the temperature control block 32 is changed at a rate of 3 ° C./sec or more by using a heating / cooling performance of each Peltier element of 15 W or more.

  By controlling each of the plurality of heating / cooling devices 51 independently, temperature locality is unlikely to occur in the temperature control block 32.

(Liquid feeding operation to reaction part 1)
The operation of feeding the fluid in the cartridge 11 to the temperature-controlled reaction unit is performed by a pressure suction mechanism (pressure suction path) formed in the liquid feed channel groove 22a of the liquid feed mechanism 12 and the temperature control block 32. 36, the pressure suction ports 41a and 41b, the pressure suction joint 31) and the pump 7 are executed through cooperative control. As described above, the reaction part is created along the shape of the reaction well 34.
FIG. 8 shows an example of the liquid feeding operation to the reaction unit.

[Step 101]
Prior to the start of liquid feeding, the membrane 21 is adhered to the surface of the cartridge 11 as shown in FIG. The space between the membrane 21 and the reaction well 34 is filled with air. In this state, when air is pressurized and introduced into the liquid feeding channel groove 22a from the pressurizing port 26, the liquid held in the membrane 21 of the liquid feeding channel groove 22a is transferred to the membrane 21 in the region facing the reaction well 34. Extruded. As a result, the membrane 21 on the reaction well 34 side is expanded and expanded as shown in FIG.

[Steps 102, 103, 104]
A control device (not shown) that manages the overall operation of the pretreatment integrated capillary electrophoresis apparatus (hereinafter referred to as “overall control unit”) confirms the amount of liquid fed at this time, and if the amount of liquid fed is a certain amount or more, (In this embodiment, if it is 15 μl or more), the pump 7 is driven and controlled, and suction (exhaust) of air in the reaction well 34 is started from the pressurized suction port 41a on the side close to the liquid inlet.

[Step 105]
The overall control unit continues the liquid feeding from the liquid feeding channel groove 22a while maintaining the suction from the pressure suction port 41a. When a certain amount or more of liquid is supplied, as shown in FIG. 9C, the membrane 21 that is performing suction is in close contact with the inner wall surface of the reaction well 34 (ie, the temperature control block 32). become. In particular, in the case of a thin reaction well 34 having a rectangular cross-sectional shape, an air pocket is formed at the corners and the degree of adhesion can be lowered when only the liquid is fed by its own weight as in the conventional apparatus. However, in this embodiment, it is possible to ensure close contact with the corners.

[Steps 106, 107, 108]
The overall control unit confirms the amount of liquid fed at this time, and if the amount of liquid fed is a certain amount or more (in this embodiment, 20 μl or more), the pressure suction port 41b far from the liquid inlet is used. Then, suction (exhaust) of air in the reaction well 34 is started.

[Steps 109, 110, 111]
The overall control unit continues the liquid feeding from the liquid feeding channel groove 22a while maintaining the suction from both the pressure suction ports 41a and 41b. When a liquid supply of a certain amount or more is further performed, the air between the temperature control block 32 and the membrane is sucked from the pressure suction port as shown in FIG. A reaction part in close contact with the entire inner wall surface (that is, the temperature control block 32) is formed.

  In the case of the present embodiment, the reaction part formed by the membrane 21 cannot expand beyond the shape of the reaction well 34. Therefore, even if the liquid feeding amount exceeds a certain amount, the temperature adjustment mechanism 12 is used. Thus, the amount of liquid for temperature control can be limited to a certain amount.

[Steps 111, 112, 113, 114]
When the liquid supply amount from the liquid supply channel 22a reaches a specified amount (25 μl or more in this embodiment), the overall control unit controls the liquid supply to end, and thereafter the suction from the pressure suction ports 41a and 41b is also ended. Then, temperature control processing (heating and cooling) is started. In FIG. 9, the suction is also finished before the temperature adjustment is started, but the suction may be continued during the temperature control.

  As shown in FIG. 9 (e), in this embodiment, the membrane 21 is firmly attached to the temperature control block 32 (without a gap), so that the contact thermal resistance between the membrane 21 and the temperature control block 32 is reduced. The heat can be efficiently supplied to or removed from the reaction solution 71 from the temperature control block 32 through the membrane 21. For example, it can be changed at a rate of 3 ° C./sec or more with respect to 25 μl of the reaction solution.

(Liquid feeding operation to reaction part 2)
By the way, the operation | movement (liquid feeding operation | movement to a reaction part) which forms a reaction part is not restricted to the suction timing shown in FIG. For example, like the suction timing shown in FIG. 10, the liquid feeding can be started after the reaction portion is formed in advance.

[Steps 201 and 202]
In this case, the overall control unit airs from the pressure suction port 41a formed in the reaction well 34 for a certain period of time (10 seconds in the present embodiment) before starting the liquid feeding from the liquid feeding channel groove 22a. The membrane 21 is partially inflated in advance. In other words, the membrane 21 is deformed so as to be in close contact with the inner wall of the reaction well 34 on the pressure suction port 41a side.

[Step 203]
Thereafter, the overall control unit controls the pump 7 to introduce air from the pressurizing port 26 of the liquid feed channel groove 22a located upstream of the reaction well 34 and hold it on the membrane 21 of the liquid feed channel groove 22a. The liquid thus fed is fed to the membrane 21 on the reaction well 34 side.

[Steps 204, 205, 206]
The overall control unit confirms the amount of liquid fed at this time, and if the amount of liquid to be fed is a certain amount or more (15 μl or more in this embodiment), the reaction well is opened from the pressurized suction port 41b far from the liquid inlet. The suction (exhaust) of the air in 34 is started.

[Step 207]
Here, the overall control unit executes the suction from the pressure suction port 41b for a certain period of time (in this example, 10 seconds), inflates the membrane 21, and completes the reaction unit. Specifically, the membrane 21 is brought into close contact with the entire inner wall of the reaction well 34 on the pressure suction port 41b side.

[Steps 208, 209, 210]
Thereafter, the overall control unit controls the pump 7 to resume the introduction of air into the pressurizing port 26 of the liquid feeding channel groove 22a located upstream of the reaction well 34, and the membrane of the liquid feeding channel groove 22a. The liquid held in 21 is sent to the membrane 21 on the reaction well 34 side.

[Steps 211 and 212]
When the liquid supply amount from the liquid supply channel groove 22a reaches a specified amount (25 μl or more in the present embodiment), the overall control unit controls the completion of liquid supply and starts temperature adjustment processing (heating and cooling).

(Liquid feeding operation to reaction part 3)
In addition, a reaction part can be formed (liquid feeding to the reaction part) using the suction timing shown in FIG. In FIG. 11, a method of simultaneously performing the liquid feeding from the liquid feeding channel groove 22a and the suction in the reaction well 34 will be described.

[Step 301]
In this case, the overall control unit performs a process of feeding the fluid by pressurizing and introducing air from the pressurizing port 26 to the liquid sending channel groove 22a, and pressurizing suction ports 41a and 41b formed in the reaction well 34. The process of sucking air and forming the reaction part is started simultaneously.

[Steps 302, 303, 304]
The overall control unit confirms the amount of liquid to be fed, and ends the liquid feeding if the amount of liquid to be fed is equal to or greater than a certain amount (25 μl or more in this embodiment). In the case of FIG. 11, the reaction part that is in close contact with the temperature control block 32 ends almost simultaneously. In the case of FIG. 11, the suction of air from both the pressure suction ports 41a and 41b is continued even after the liquid feeding is finished. As in the case of the suction timing described above, when the close contact between the membrane 21 and the temperature control block 32 is secured at the end of the liquid feeding, at this point, both the pressure suction ports 41a and 41b are used. The suction may be terminated.

[Steps 305, 306, 307]
The overall control unit starts temperature adjustment processing (heating and cooling), and when the temperature sensor 33 confirms that the temperature of the liquid in the reaction well 34 has reached 90 ° C. or higher, the pressure suction ports 41a and 41b End the suction of air from both. In FIG. 11, only the case of heating is assumed, but of course, the process of cooling the temperature of the liquid in the reaction well 34 to a predetermined temperature is also executed.

(Liquid feeding operation from the reaction part)
FIG. 12A shows the state of the reaction part at the time when the reaction process in the reaction well 34 is completed. At this point, the membrane 21 in close contact with the reaction well 34 is filled with fluid. Next, the overall control unit sucks air from the suction port 25 of the liquid feeding channel groove 22b located on the downstream side of the reaction well 34, and pulls down the membrane 21 at a position facing the liquid feeding channel groove 22b (expansion). ) As a result, the fluid retained on the membrane 21 on the reaction well 34 side is sucked out to the liquid feed channel groove 22b side. At the same time, the overall control unit introduces (pressurizes) air into the pressure suction ports 41 a and 41 b formed in the reaction well 34. This state is shown in FIG. It should be noted that the introduction of air into the reaction well 34 does not have to be performed by both the pressurized suction ports 41a and 41b, and only one of them may be used. In any case, air is introduced into the reaction well 34 to pressurize it, and the membrane 21 is deformed so as to push it up, so that the membrane 21 is surely returned to the state of being in close contact with the surface of the cartridge 11. That is, the reaction liquid can be reliably discharged from the reaction part. As a result, the liquid can be sent reliably and in a short time compared with the liquid sending only by the suction from the liquid sending channel groove 22b.

[Example 2]
In the above-described embodiment, the heat insulating material 35 is disposed so as to surround the reaction well 34 (that is, the heat insulating material 35 is disposed over the entire side surface of the temperature control block 32) to reduce heat radiation from the reaction portion to the outside. The method was explained. Furthermore, in order to reduce the amount of heat radiation, the portion of the main body of the cartridge 11 that opposes the reaction region forming region may be made thinner than the other portions, and the heat insulating material 35 may be installed in the thinned portion. If such a structure is adopted, heat radiation from the upper surface side of the reaction portion to the cartridge 11 can be further reduced.

  More positively, not only the heat insulating material but also a heating device or a heating / cooling device may be provided in a region facing the reaction portion. If heating or heating / cooling is performed from the upper surface of the reaction section, the temperature change of the reaction solution can be further accelerated. FIG. 13 shows an example in which the heating device 62 is installed on the upper surface side of the cartridge 11. In FIG. 13, an air layer 63 is formed in a portion of the cartridge 11 that faces the reaction portion, and the heat insulation effect is also enhanced. However, the heating device 62 may be brought into direct contact with the upper surface of the cartridge 11 having a structure without the air layer 63. When the heating device 62 is brought into direct contact with the surface of the cartridge 11, it is desirable that the set temperature of the heating device 62 matches the target heating temperature. As shown in FIG. 13, when the air layer 63 is sandwiched between the heating device 62 and the reaction section, the temperature of the sandwiched air is sufficiently increased by setting the temperature higher than the target heating temperature. It needs to be heated.

  At the time of cooling, processing and operations such as stopping heating of the heating device 62 or removing the heating device 62 from the upper surface of the cartridge 11 are performed.

  By the way, when the heating device 62 (including the heating / cooling device here) is provided on the upper surface of the cartridge 11 as in this embodiment, the heating device 62 is controlled by heating the heating / cooling mechanism in the temperature control mechanism 13. It is also desirable to synchronize with the cooling control. If heat exchange is performed from both the upper surface side and the lower surface side of the reaction section, the temperature change of the reaction solution is further promoted, and the temperature difference between the upper surface and the lower surface of the reaction solution is less likely to occur.

(Other examples)
The present invention is not limited to the above-described embodiments, but includes various modifications. For example, the pressure suction ports 41a and 41b are provided on the bottom surface of the reaction well 34 having a rectangular cross section. However, the pressure suction ports 41a and 41b may be provided at any position, for example, on the side surface, the bottom surface, or the corner, as long as the corner is formed by the side surface and the bottom surface. Can do. In the above-described embodiment, the case where both of the pressure suction ports 41a and 41b can be used for suction as well as pressurization has been described. An opening may be provided, and a pressure-dedicated tube and a suction-dedicated tube may be provided in the temperature control block 32. For example, the pressure suction port 41a and its connection pipe may be used exclusively for suction, and the pressure suction port 41b and its connection pipe may be used exclusively for pressurization.

  In the above-described embodiment, the case where the cross-sectional shape of the reaction well 34 is a rectangular shape has been described. However, the reaction well 34 may not necessarily have a rectangular shape. For example, the corner portion may be rounded, or the cross-sectional shape of the reaction well 34 in the liquid feeding direction may be elliptical.

  Moreover, in order to explain this invention in an easy-to-understand manner, the above-described embodiments are described in detail for some of the embodiments, and it is not always necessary to have all the configurations described. Further, a part of a certain embodiment can be replaced with the configuration of another embodiment, and the configuration of another embodiment can be added to the configuration of a certain embodiment. Moreover, it is also possible to add, delete, or replace another configuration for a part of the configuration of each embodiment.

1: Pretreatment integrated capillary electrophoresis apparatus 2: Pretreatment unit 3: Analysis unit 4: Autosampler 5: Capillary 6: Detection unit 7: Pump 8: Oven 9: High voltage power supply 11: Cartridge 12: Liquid feeding mechanism 13: Temperature control mechanism 21: Membrane 22a, 22b: Liquid supply channel 23a, 23b: Narrow channel 24a, 24b: Well 25: Suction port 26: Pressurization port 31: Joint for pressure suction 32: Temperature control block 33: Temperature sensor 34: reaction well 35: heat insulating material 36: pressure suction path 41a, 41b: pressure suction port 51: heating / cooling device 52: heat radiation block 53: heat sink 54: air cooling fan 55: heat radiation part 61: temperature control board 62: Heating device 63: Air layer 71: Reaction liquid

Claims (13)

For forming a reaction part that faces the open end of a flow path formed on the biochemical cartridge side across the membrane when a biochemical cartridge having a membrane that is an elastic body deformable by air pressure is attached to the surface. A concave structure having a rectangular cross section, a first opening and a second opening provided near a corner of the concave structure, a first conduit connected to the first opening, and the second opening A temperature control block formed with a second conduit connected to
At the time of air suction, air is sucked from the concave structure with a time difference from the first pipe line and the second pipe line, while at the time of air introduction, the air passes through the first pipe line and the second pipe line. A temperature control mechanism for a biochemical cartridge, comprising: a pump for simultaneously pressurizing the concave structure . For forming a reaction part that faces the open end of a flow path formed on the biochemical cartridge side across the membrane when a biochemical cartridge having a membrane that is an elastic body deformable by air pressure is attached to the surface. A concave structure, a first opening for air suction provided on the inner peripheral surface of the concave structure, a first conduit connected to the first opening, and an inner peripheral surface of the concave structure A temperature control block formed with a second opening for introducing air and a second pipe connected to the second opening ;
A first pump for sucking air from the concave structure through the first conduit;
Second pump and biochemical cartridge temperature adjusting mechanism having to introduce air into the concave structure through the second conduit. The temperature control mechanism for a biochemical cartridge according to claim 1 or 2 ,
The temperature control mechanism for a biochemical cartridge, wherein the temperature control block has a heat insulating structure around the concave structure. The temperature control mechanism for a biochemical cartridge according to claim 1 or 2 ,
A biochemical cartridge temperature control mechanism comprising a heat insulating structure on an upper surface side of the biochemical cartridge. The temperature control mechanism for a biochemical cartridge according to claim 1 or 2 ,
A temperature control mechanism for a biochemical cartridge, comprising a heating device or a heating / cooling device on an upper surface side of the biochemical cartridge. The temperature control mechanism for a biochemical cartridge according to claim 1,
The biochemical cartridge temperature control mechanism, wherein the pump sucks air from the first opening and the second opening after the liquid feeding into the concave structure is started. The temperature control mechanism for a biochemical cartridge according to claim 1,
The pump sucks air from the first opening and the second opening prior to the start of liquid feeding into the concave structure. The temperature control mechanism for a biochemical cartridge according to claim 1,
The temperature control mechanism for a biochemical cartridge, wherein the pump sucks air from the first opening and the second opening simultaneously with the start of liquid feeding into the concave structure. The temperature control mechanism for a biochemical cartridge according to claim 1,
The temperature control mechanism for a biochemical cartridge, wherein the pump introduces air from the first opening and the second opening during liquid feeding from the concave structure. For forming a reaction part that faces the open end of a flow path formed on the biochemical cartridge side across the membrane when a biochemical cartridge having a membrane that is an elastic body deformable by air pressure is attached to the surface. A concave structure with a rectangular cross section ,
A first opening and a second opening provided near a corner of the concave structure;
A first conduit connected to the first opening ;
A temperature control block having a second conduit connected to the second opening . For forming a reaction part that faces the open end of a flow path formed on the biochemical cartridge side across the membrane when a biochemical cartridge having a membrane that is an elastic body deformable by air pressure is attached to the surface. The concave structure of
A first opening for air suction provided on the inner peripheral surface of the concave structure;
A first conduit connected to the first opening;
A second opening for air introduction provided on the inner peripheral surface of the concave structure;
A second conduit connected to the second opening;
Temperature control block with. A biochemical cartridge in which a membrane, which is an elastic body that can be deformed by air pressure, is attached to the surface;
When the biochemical cartridge is mounted, a concave structure having a rectangular cross section for forming a reaction portion facing the opening end of a flow path formed on the biochemical cartridge side with the membrane interposed therebetween, and near the corner of the concave structure a first opening and a second opening provided, the first and conduit, the second second temperature control of conduits and are formed which are connected to an opening which is connected to the first opening When sucking air, the block sucks air from the concave structure with a time lag from the first pipe line and the second pipe line, whereas when introducing air, the first pipe line and the second pipe line are sucked . A temperature control mechanism for a biochemical cartridge having a pump for simultaneously pressurizing the concave structure through a conduit ;
A biochemical processing apparatus, comprising: a nucleic acid analyzer that introduces a fluid from the biochemical cartridge and analyzes a nucleic acid contained in the fluid. A biochemical cartridge in which a membrane, which is an elastic body that can be deformed by air pressure, is attached to the surface;
At the time of mounting the biochemical cartridge, a reaction structure forming concave structure facing the opening end of the flow path formed on the biochemical cartridge side with the membrane interposed therebetween, and air provided on the inner peripheral surface of the concave structure A first opening for suction; a first conduit connected to the first opening; a second opening for air introduction provided on an inner peripheral surface of the concave structure; and the second opening. A temperature control block formed with a second pipe connected to the first pump, a first pump for sucking air from the concave structure through the first pipe, and the concave structure through the second pipe A temperature control mechanism for a biochemical cartridge having a second pump for introducing air into the
A nucleic acid analyzer for introducing a fluid from the biochemical cartridge and analyzing a nucleic acid contained in the fluid;
A biochemical processing apparatus.

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