JP2002058470A - Apparatus and method for temperature control for minute chemical device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a temperature control apparatus for a minute chemical device capable of readily and accurately regulating a temperature of a fixed minute area in a minute chemical device generally having difficulty in an accurate temperature measurement, and a method for temperature control. SOLUTION: The temperature control apparatus for a minute chemical device has one or more temperature control blocks set in a temperature range of a desired set temperature ±10 deg.C, which are brought into contact with both the outsides of the obverse and reverse of a filmy or laminar minute chemical device having a capillary channel and one or more desired set areas. The temperature control method for a minute chemical device comprises bringing an object set at a desired set temperature ±10 deg.C into contact with both the outsides of the obverse and reverse of the filmy or laminar minute chemical device.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention is used in a wide range of fields such as the chemical industry, biochemical industry, agriculture, forestry, fisheries, medical care, food industry, pharmaceutical industry, environmental protection, criminal investigation, sports, etc. A microchemical device having a microchannel in a member, for example, a microreaction device (microreactor) for chemistry, biochemistry, etc., an integrated DNA analysis device, a microelectrophoresis device, a microchromatography device, a micromembrane separation device The present invention relates to a temperature control device and a temperature control method for a microanalytical device such as chemistry and biochemistry such as a microextraction device.

More particularly, the present invention relates to an apparatus and a method for controlling the temperature of a plate-like or film-like microchemical device, and more particularly to a method and an apparatus for accurately controlling the temperature of a minute region of a microchemical device. Among them, it is useful for microreactors, especially for PCR (PCR: polymerase chain reaction, hereinafter referred to as PCR) devices.

[0003]

2. Description of the Related Art As a microchemical device, a thin groove-like flow path is formed on a base material such as silicon, quartz, glass, or a polymer by an etching method, and a cover is attached as necessary to form a groove-like flow path. It is known that the channel is formed as a capillary channel, and is used as a liquid channel, a channel-shaped reaction tank, a gel channel for separation, etc., and a mechanism for heating a specific region of such a microchemical device. It has been reported how to incorporate an electric heater (eg, "Science",
1998, vol. 282, p. 484).

[0004] However, it is extremely difficult to accurately control the temperature of a minute portion, and the difference between the set temperature and the actual temperature tends to be large. The first is the difficulty in accurately detecting the temperature of a minute part, and the second is that the surface area of the part to be temperature-regulated is very large compared to the volume of the part, so that heat is not generated. It was difficult to control the temperature stably due to large ingress and egress.

[0005]

SUMMARY OF THE INVENTION The problem to be solved by the present invention is that a small desired region in a small chemical device, which is generally difficult to accurately measure temperature, can be easily and accurately adjusted in temperature. An object of the present invention is to provide a temperature control device for a chemical device and a temperature control method.

[0006]

Means for Solving the Problems As a result of diligent studies on a method for solving the above-mentioned problems, the present inventors have found that temperature is separately adjusted on both sides of a portion of a plate-like or film-like microchemical device to be temperature-controlled. Contact with the substance
The above problems have been solved, and it has been found that accurate temperature control of minute parts is possible, and the present invention has been completed.

That is, the present invention provides (1) a desired set temperature of ± 10 ° C. which is in contact with both front and back outer surfaces of a film-shaped or plate-shaped microchemical device having a capillary flow path and one or more set desired temperature regions. A temperature control device for a microchemical device having one or more temperature control blocks set in a temperature range of

(2) The contact area between the temperature control block and the microchemical device is 1 × 10 ⁻⁸ m ² to 1 × 10 ^−4.
is m ² and the temperature adjusting device for fine chemical device according to (1),

(3) The temperature controller for a microchemical device according to (1), wherein the desired temperature range to be set is a part of the microchemical device.

(4) The temperature control for a microchemical device according to any one of (1) to (3), wherein the temperature control block is in contact with a plurality of set desired temperature regions of the same or different temperatures of the microchemical device, respectively. Equipment and

(5) The thickness of the microchemical device is 10
a temperature controller for a microchemical device according to any one of (1) to (4),

(6) The microchemical device according to any one of (1) to (5), wherein the microchemical device is a microreactor;

(7) The microreactor is PC
A temperature control device for a microchemical device according to (6), which is an arel (PCR) device;

(8) The temperature range of the set desired temperature ± 10 ° C. is set on both the front and back outer surfaces of the film-shaped or plate-shaped microchemical device having the capillary channel and one or more set desired temperature regions. A method for controlling the temperature of a microchemical device for contacting an object,

(9) The method for adjusting the temperature of a microchemical device according to (8), wherein the desired temperature range to be set is a plurality of regions having the same or different temperatures.

(10) The thickness of the microchemical device is 1
(8) or (9), wherein the temperature of the microchemical device is from 0 μm to 10 mm;

(11) The method for controlling the temperature of a microchemical device according to any one of (8) to (10), wherein the microchemical device is a microreactor; and

(12) The method for controlling the temperature of a microchemical device according to (11), wherein the microreactor is a PC device (PCR) device.

[0019]

DESCRIPTION OF THE PREFERRED EMBODIMENTS In the present invention, a temperature control (hereinafter, referred to as "temperature control") is described.
The microchemical device to be temperature-regulated may be simply referred to as temperature control) has a capillary channel in a film or plate member, and has a thickness of 10 μm to 10 μm.
mm, preferably 5 mm or less, most preferably 2 mm or less.
mm or less. If the thickness is larger than this, the temperature control accuracy tends to decrease when the desired setting temperature region is minute. The lower limit of the thickness is not particularly limited from the viewpoint of temperature control accuracy, but is preferably 10 μm or more, more preferably 50 μm or more, from the viewpoint of difficulty in manufacturing the device.

The distance (the smaller the depth from the front surface or the back surface) of the capillary channel in the desired temperature region of the microchemical device from the outer surface of the device is preferably 3 mm or less, more preferably. Is 1 mm or less, most preferably 0.5 mm or less. If the distance is longer than this, especially when an exothermic reaction or an endothermic reaction occurs in the flow path, when the temperature difference between the desired setting temperature region and its peripheral portion is large, or when the desired setting temperature region is minute, etc. Temperature control accuracy tends to decrease. The lower limit of the distance does not need to be limited in terms of temperature control accuracy, but is preferably 2 μm or more, and more preferably 5 μm or more, in view of the difficulty in manufacturing the device.

The material constituting the microchemical device to be temperature-controlled is arbitrary, and may be silicon, quartz, glass, an organic polymer, or the like. However, the organic polymer has a low thermal conductivity. This is preferable because minute temperature regions having different temperatures can be easily formed in the device, and the effects of the present invention can be easily exerted.

The apparatus and method of the present invention are for adjusting the temperature of a desired set temperature range provided in a microchemical device. One or more desired setting temperature regions are provided in the microchemical device whose temperature is to be adjusted, and the flow path in the region is adjusted to the desired setting temperature.

The desired setting temperature range may be the entire microchemical device, or the device may have a region other than the desired setting temperature range. The desired setting temperature region may be a plurality of regions having the same temperature or different temperatures. The present invention is particularly effective when the device has a region other than the desired setting temperature region, or when the desired setting temperature region is a plurality of regions. From this point, the microchemical device to which the present invention is applied is suitable for a microreactor, and particularly suitable for a PCR device.

According to the present invention, the desired desired temperature is set to ± 10 ° from the front and back outer surfaces of the microchemical device within the desired desired temperature range.
C., preferably by contacting an object set within a temperature range of the desired set temperature ± 5 ° C., more preferably the desired set temperature ± 1 ° C., so that the set desired temperature region is adjusted to the set desired temperature. The temperature of the object may be the same on the front and back of the microchemical device, or may be different, as long as they are within the above ranges, but preferably the same.

The temperature of the object is determined by the thickness of the microchemical device, the type of reaction performed in the flow path (for example, exothermic reaction or endothermic reaction), the temperature difference between the set desired temperature region and its peripheral portion, and the set desired temperature region. The set value can be changed in the above-mentioned temperature range depending on the size and the like. When the thickness of the microchemical device is small, it is preferable to set the desired temperature to ± (1 ° C. or less).

The temperature-controlled object brought into contact with the microchemical device is arbitrary, and may be a solid, a liquid, a gas, a mist, or the like. A solid (hereinafter, a temperature-controlled solid is referred to as a temperature-control block). It is preferable that the temperature control block has any shape, including, for example, a film shape) because of its simple structure, and it is more preferable that the temperature control block is made of metal because of its good thermal conductivity. The type of the object to be contacted may be different on the front and back of the microchemical device, but is preferably the same.

When the temperature controlled object to be brought into contact with the microchemical device is a temperature control block, the contact area between the temperature control block and the microchemical device is 1 × 10 ⁻⁸ m.
It is preferably at least ^{2 and} more preferably at least 1 × 10 ⁻⁷ m ² . If the contact area is too small,
High-precision temperature control becomes difficult.

The lower limit of the preferable contact area also depends on the thickness of the microchemical device and the distance from the surface of the microchemical device to the flow path. The smaller the thickness of the microchemical device, the smaller the distance from the surface to the flow path. The more accurate the temperature control becomes, the smaller the area becomes. The upper limit of the contact area does not need to be limited in terms of temperature control accuracy, but in order to exhibit the characteristics of the microchemical device, each contact area is 1 × 1.
It is preferably 0 ^-4 m ² or less, and more preferably 1 × 10 ^-5 m ² or less.

The method of controlling the temperature of the temperature-controlled object to be brought into contact, for example, the temperature control block is arbitrary. For example, the temperature can be controlled by an electric heater; the temperature can be controlled by a Peltier element; Temperature control by controlling the flow rate of cold air such as liquefied carbon dioxide gas or liquid nitrogen; temperature control by electromagnetic waves such as infrared rays and microwaves; temperature control by fluctuating magnetic fields; and a method of combining these. Needless to say, the temperature control mentioned here includes not only adjustment to a constant temperature, but also adjustment not to deviate from a certain temperature range, temperature increase, temperature decrease, and other programmed temperature adjustments.

The temperature of the object to be brought into contact may be adjusted based on the measured value of the temperature of the microchemical device.
It is preferable that the temperature of the object itself is controlled without detecting the temperature of the microchemical device.
Therefore, in the present invention, it is not necessary to detect the temperature of the microchemical device.

The contact with the microchemical device includes not only a fixed contact state but also contact at a specific time for a specific time. The contact may be through oil, grease, or the like to increase thermal conductivity. An object of the present invention is to bring a temperature-controlled object, for example, a temperature control block, into contact with a set desired temperature range of the microchemical device from both front and back outer surfaces of the microchemical device.

Therefore, it is sufficient that the desired temperature range of the microchemical device is sandwiched between the temperature control objects from the front and back.
The other regions are arbitrary, and for example, a temperature-controlled object may be in contact with one surface. However, it is preferable that only the desired setting temperature range of the microchemical device is sandwiched by the temperature-controlled objects.

That is, the shape of the contact portion of the temperature-controlled object with the microchemical device is preferably mirror-symmetrical on both sides. Of course, the temperature-controlled object may have a portion that deviates from the microchemical device (a portion that does not contact the microchemical device), and the shape of this portion is arbitrary. In a similar sense, the shape of the temperature-regulated object that comes into contact with a portion of the microchemical device where no flow path exists does not matter.

There may be a plurality of desired setting temperature ranges for performing the temperature control. Set desired temperatures for multiple set desired temperature areas
The temperature may be the same or different. In this case, the same applies to each temperature region. The plurality of desired temperature ranges may be in contact with each other or may be independent.

The temperature range other than the desired temperature range for the setting of the microchemical device is arbitrary. For example, an object whose temperature is controlled on one side may be contacted, released to room temperature air, Although it may be exposed to an air current, it is preferable to release to room temperature air or to expose to an air current of room temperature air.

The method of bringing the temperature control device into contact with the front and back outer surfaces of the microchemical device is arbitrary. For example, the microchemical device may be sandwiched by one set of independent temperature control devices, or one set of temperature control devices may be used. The control devices may be connected by hinges to sandwich the microchemical device. Alternatively, the temperature control block may have a U-shaped integral structure that can be sandwiched between the temperature control objects from the front and back.

[0037]

EXAMPLES The present invention will be described in more detail with reference to the following Examples, but it should not be construed that the present invention is limited thereto.

[Example 1] [Preparation of microchemical device] Average molecular weight of about 2000
Trifunctional urethane acrylate oligomer ("Unidick V-426" manufactured by Dainippon Ink and Chemicals, Inc.)
3 "), 35 parts of 1,6-hexanediol diacrylate (" New Frontier HDDA "manufactured by Daiichi Kogyo Seiyaku Co., Ltd.), and nonylphenoxypolyethylene glycol (n = 17) acrylate (Daiichi Kogyo Seiyaku Co., Ltd.) 30 parts of “N-177E” manufactured by the company, 5 parts of 1-hydroxycyclohexylphenyl ketone (“Irgacure 184” manufactured by Ciba Geigy), and 2,4-diphenyl-4-methyl-1-pentene (Kanto) (Chemical Co., Ltd.) in an amount of 0.1 part to prepare a homogeneous composition (x-1).

The composition (x-1) was applied to a 5 cm × 5 cm × 1 mm acrylic resin plate (“Acrylite L” manufactured by Mitsubishi Rayon Co., Ltd.) using a 127 μm bar coater. Then, the coating film was semi-cured by irradiating ultraviolet rays of 100 mw / cm ² for 3 seconds to form a semi-cured resin first layer (2) having a thickness of 85 μm.

On this first resin layer (2), 127 μm
The composition (x-1) was applied using a bar coater, and 100 mw / cm ² of ultraviolet light was applied to portions other than the portion serving as the flow path (4) shown in FIG. 1 using a photomask. 1
Irradiation for 2 seconds, after semi-cured coating of the irradiated part, remove the uncured composition of the unirradiated part with a water stream, width 92μm,
Channel (4) having a depth of 88 μm and a length of 18 mm × 20 turns
The resin second layer (3) in which the groove to be formed was formed as a resin defective portion was formed.

Thickness 30 μm with one surface subjected to corona discharge treatment
Of a biaxially stretched polypropylene film (OPP film manufactured by Nimura Chemical Co., Ltd.) (not shown) using a 127 μm bar coater on the surface of the composition (x-
1) is applied, and the same ultraviolet ray as used above is irradiated for 1 second without a photomask, and the coating film is semi-cured to a thickness of 98 μm.
And the semi-cured coating film is brought into close contact with the resin second layer (3) on the substrate (1), and in this state, ultraviolet rays are further irradiated for 30 seconds to remove all the resin layers. After curing and bonding, the polypropylene film (not shown) is peeled off to form the third resin layer (5), thereby forming the second resin layer (3).
Was formed into a capillary channel (4).

Next, at the position of one end of the flow path (4),
A hole was made in the base material (1) and the first resin layer (2) with a 1.6 mm drill, and a 1.6 mm diameter stainless steel pipe was bonded to the hole to form an inlet (6). In the same manner, an outlet (7) was formed at the other end of the flow channel, and a microchemical device for PCR [D-1] having the shape shown in FIGS. 1 and 2 was produced.

[Preparation of Temperature Control Device] Three 6 mm × 6 mm
As shown in FIGS. 3 and 4, a 1 mm × 7 mm × 60 mm acrylic plate was arranged and fixed as a spacer (12), and a brass square bar (11) having a size of 60 mm was fixed to the brass square bar (11). 1.7m diameter on 6mm x 6mm end face
A 30 mm deep drill hole was drilled, and a sheath-type thermocouple (13) was attached as a temperature sensor for temperature control.

Further, each of the brass square bars (11)
A polyimide resin-filled electric heater (14) was attached to the bottom surface of mm × 60 mm. And three sets of thermocouples (1
3) and the electric heater (14) were each connected to a temperature controller (not shown) to prepare a temperature controller having three temperature control blocks (11). Further, another one of the same temperature control devices was manufactured to form a set of temperature control devices.

[PCR test] Template plasmid DNA: 4.0 μm
l, polymerase [TaKaRa Ex Taq T manufactured by Takara Shuzo Co., Ltd.
M ”]: 2.0 μl, buffer [Takara Shuzo Co., Ltd.“ 10X ExTaq ”
TM Buffer ”]: 8.0 μl, substrate [Takara Shuzo Co., Ltd.“ d
NTP Mixture (2.5 mM each) "]: 6.4 μl, primer
[Takara Shuzo Co., Ltd. "Fluorescein-Labeled Primer M4
(1 pmol / μl)]]: 20 μl, primer [Takara Shuzo Co., Ltd. “Fluorescein-Labeled Primer RV-M (1 pmol / μl)”]:
20 μl and 19.6 μl of sterile distilled water were mixed to prepare a reaction solution, and the mixture was shaken in a suction bell and shaken with an aspirator.
The pressure was reduced to 0 seconds to remove dissolved air, and the atmosphere was sealed with argon.

The three temperature control blocks (11) of the temperature control device are controlled at 95 ° C., 75 ° C. and 45 ° C., respectively, and the desired temperature range (region α) of the microchemical device set at 95 ° C. is controlled at 95 ° C. In the block, the desired temperature region (region β) set at 75 ° C. was sandwiched by a temperature control block of 75 ° C., and the desired temperature region (region γ) set at 45 ° C. was sandwiched by a temperature control block of 45 ° C.

At room temperature 25 ° C., the PCR reaction solution was introduced from the inlet (6) of the microchemical device (D-1) using a microsyringe at a flow rate of a residence time of 20 minutes.
The PCR test was carried out for a period of one minute. The reaction solution flowing out of the outlet (7) was collected and subjected to electrophoresis analysis.
It was confirmed that the DNA was amplified. Same PCR
When the test was carried out at 15 ° C. and 35 ° C. while changing the room temperature, the obtained electrophoresis results were the same, and were not affected by the room temperature, and reproducible results were obtained.

[Comparative Example 1] An acrylic resin plate having a thickness of 1 mm ("Acrylite L" manufactured by Mitsubishi Rayon Co., Ltd.) instead of a temperature control device to be brought into contact with the third resin layer (5) of a microchemical device The temperature control device, which is made by fixing the cover made of a resin and contacting the substrate (1) surface of the microchemical device, with the resin second layer (the center of the regions α, β, and γ of the microchemical device) The temperature of the temperature control block (11) was adjusted so that the display temperatures of the thermocouple (not shown) 3) and the resin third layer (5) were shaved and embedded were 95 ° C, 75 ° C, and 45 ° C, respectively. A PCR test was performed at room temperature of 15 ° C., 25 ° C., and 35 ° C. in the same manner as in Example 1 except for the above.

As a result, at 15 ° C., the solution boiled in the region α, causing a liquid breakage, and almost no DNA was observed in the effluent. At room temperature of 25 ° C. and 35 ° C., although DNA amplification was observed, the concentration was lower than that of Example 1. As described above, in the temperature control from one side by the thermocouple embedded in the microchemical device, it can be seen that there is a difference between the display temperature and the actual temperature, and the temperature is easily affected by the fluctuation of the room temperature.

[0050]

Industrial Applicability The present invention relates to a temperature control device for a microchemical device, which can easily and accurately control the temperature of a microscopic desired area in a microchemical device for which accurate temperature measurement is generally difficult, and a temperature control. Methods can be provided, especially microreactors and microPCR
It is suitably used for a temperature control device for a device and a temperature control method.

[Brief description of the drawings]

FIG. 1 is a schematic diagram of a plan view of a microchemical device for PCR used in an example viewed from a direction perpendicular to the surface.

FIG. 2 is a schematic view of an elevation view of a microchemical device for PCR used in Examples.

FIG. 3 is a plan view of one of a pair of symmetric temperature control devices manufactured in the example.

FIG. 4 is a side view of one side of a set of symmetric temperature control devices manufactured in the example.

[Explanation of symbols]

 1: substrate 2: resin first layer 3: resin second layer 4: flow path 5: resin third layer 6: inlet 7: outlet 11: temperature control block (square bar made of brass) 12: spacer 13: Sheath type thermocouple 14: Polyimide resin embedded heater α: Desired temperature region set at 95 ° C β: Desired temperature region set at 75 ° C γ: Desired temperature region set at 45 ° C

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) G01N 37/00 101 G01N 37/00 101 F term (Reference) 4B029 AA23 BB20 CC01 4G075 AA13 AA32 AA63 AA65 BA02 BA05 BD14 BD16 BD26 CA02 CA03 CA13 CA26 CA42 CA63 DA02 DA18 EA06 EB46 EE12 EE23 EE31 FA20 FB01 FB02 FB06 FB11 FB12 FC11

Claims (12)

[Claims] 1. A temperature range of a set desired temperature ± 10 ° C., which is in contact with both front and back outer surfaces of a film-shaped or plate-shaped microchemical device having a capillary flow path and one or more set desired temperature regions. A temperature control device for a microchemical device having one or more temperature control blocks. 2. The microchemical device temperature controller according to claim 1, wherein the contact area between the temperature control block and the microchemical device is 1 × 10 ⁻⁸ m ² to 1 × 10 ⁻⁴ m ² . 3. The temperature controller for a microchemical device according to claim 1, wherein the desired setting temperature region is a partial region of the microchemical device. 4. The micro-chemical device temperature control device according to claim 1, wherein the temperature control block is in contact with a plurality of desired temperature ranges of the same or different temperatures of the micro-chemical device. 5. The microchemical device has a thickness of 10 μm.
The temperature controller for a microchemical device according to any one of claims 1 to 4, wherein the temperature is 10 to 10 mm. 6. The temperature controller for a microchemical device according to claim 1, wherein the microchemical device is a microreactor. 7. The temperature control device for a microchemical device according to claim 6, wherein the microreactor is a PCR (PCR) device. 8. An object set in a temperature range of set desired temperature ± 10 ° C. on both front and back outer surfaces of a film-shaped or plate-shaped microchemical device having a capillary flow path and one or more set desired temperature regions. Temperature control method of a microchemical device for contacting the surface. 9. The method for adjusting the temperature of a microchemical device according to claim 8, wherein the desired setting temperature region is a plurality of regions having the same or different temperatures. 10. The microchemical device has a thickness of 10 μm.
The temperature control method for a microchemical device according to claim 8, wherein the diameter is from 10 to 10 mm. 11. The method for controlling the temperature of a microchemical device according to claim 8, wherein the microchemical device is a microreactor. 12. The method for controlling the temperature of a microchemical device according to claim 11, wherein the microreactor is a PC device (PCR).