JP5540705B2 - Temperature control device, temperature control method, and program thereof - Google Patents

Description

The present invention relates to a temperature control device, a temperature control method, and a program thereof.
This application claims priority on December 07, 2007 based on Japanese Patent Application No. 2007-317159 for which it applied to Japan, and uses the content here.

  Polymerase chain reaction (PCR) exists as a technique for amplifying deoxyribonucleic acid (DNA). This PCR method is a technique capable of amplifying DNA in a short time by periodically raising and lowering the temperature of an aqueous solution containing DNA. By the way, in order to accurately amplify an arbitrary fragment of DNA using this PCR method, temperature control in an aqueous solution must be strictly performed.

Here, Patent Literature 1 and Patent Literature 2 are disclosed as temperature estimation techniques. The technique of Patent Document 1 is a gas temperature estimation method for controlling a combustion path of a waste incinerator, particularly a fluidized bed furnace. Based on this method, the amount of air supplied to the fluidized bed passage having a combustion time of several tens of seconds is controlled. In this method, a thermocouple having a time delay of temperature measurement of about 1 to 2 minutes is used, and a gas in which the measurement delay due to the thermocouple is compensated by the Kalman filtering unit based on the measured temperature of the thermocouple calculated by the system calculation process. Estimate the temperature and match the combustion state of the waste incinerator.
Further, the technique of Patent Document 2 is a method for measuring a subject's body temperature and approximating a temperature measurement value with an S-shaped curve, for example, a logistic curve, thereby obtaining a blood glucose concentration with high accuracy without collecting blood from the subject. It is.
Japanese Patent No. 3424024 Japanese Patent No. 3874743

  By the way, in the estimation of the temperature in the aqueous solution in the above-mentioned PCR method, it is necessary to consider the characteristics of the measurement object. That is, in the measurement of the temperature in the aqueous solution in the PCR method, the measurement area of the temperature measurement object is a small cavity (small space into which the aqueous solution is injected), and the measurement object is small. Moreover, in the temperature measurement by the conventional technique (for example, thermography), only the surface temperature of the object is measured, and the temperature in the vicinity of DNA in the aqueous solution is measured as in the PCR method. The internal temperature of an object cannot be estimated. It is also conceivable to measure the temperature inside the aqueous solution by installing a thermometer such as an ultrafine thermocouple in the cavity, but placing a thermocouple in an aqueous solution containing a substance that is vulnerable to heat called DNA It is not preferable.

  Accordingly, an object of the present invention is to provide a chip for chemical reaction, a temperature control device, a temperature control method, and a program thereof that can estimate the internal temperature of a measurement object containing a small and brittle substance from the external temperature. It is one.

(1) In order to achieve the above object, one embodiment of the present invention includes a lower plate portion, an upper plate portion joined to a surface of the lower plate portion, and between the lower plate portion and the upper plate portion. A chip for chemical reaction used for a chemical reaction of a reactant contained in the cavity by heating and cooling from the outside, and measuring a temperature of a lower external contact of the cavity The chemical reaction chip comprised of a lower temperature measuring element and an upper temperature measuring element for measuring the temperature of the upper external contact of the cavity in the vertical direction from the lower temperature measuring element of the upper plate part, is housed in the chip a temperature control unit for controlling the temperature of repeating the heating and the cooling of the reaction product, is connected to the lower temperature measuring element and the upper temperature measuring element, it is housed in the chemical reaction chip using a temperature estimation algorithm Was Bei example a temperature estimating section, the performing temperature estimation of the reactants, the temperature estimation unit by using the temperature estimation algorithm, the time series of the lower temperature measuring element and the upper temperature measurement measurement results obtained from the device The frequency component is obtained by Fourier transforming each time-series data when repeating heating and cooling from the outside to the cavity of the chemical reaction chip, and using the frequency component, the chemical reaction chip It is a temperature control apparatus characterized by estimating the time series temperature inside .

(2) The above temperature control device may be configured as follows: the temperature estimation unit uses a temperature estimation algorithm, and a measurement result obtained from the lower temperature measurement element and the upper temperature measurement element Each of the time series data when the heating and cooling from the outside to the cavity of the chemical reaction chip are repeated by Fourier transform to obtain frequency components, the upper external contact and the lower side The thickness d1 of the upper plate portion on the straight line connecting the external contacts, the thickness d2 obtained by subtracting the thickness d1 from the distance between the predetermined temperature estimation point in the cavity and the upper external contact point, the thickness of the lower plate portion A transfer matrix of temperature connecting the thickness d1 and the thickness d2 when the thickness d3 is obtained by subtracting the thickness d4 of the plate portion from the distance between the temperature estimation point and the lower external contact; Thickness d2 and thickness a temperature transfer matrix connecting d3, a temperature transfer matrix connecting thickness d3 and thickness d4, a temperature transfer matrix connecting thickness d1 to thickness d3, and thickness d1 to thickness d4. The temperature transfer point matrix and the frequency components obtained by Fourier transforming the time series data of the measurement results obtained from the lower temperature measurement element and the upper temperature measurement element, respectively, are used to calculate the temperature of the temperature estimation point. presume.

(3) The above temperature control apparatus may be configured as follows: based on the temperature estimated value obtained by the temperature estimating unit and the desired temperature of the reactant recorded in advance, the temperature estimation A temperature control unit is further provided for performing temperature control of heating and cooling so that the value approaches the desired temperature.

(4) Moreover, another one aspect | mode of this invention is a cavity provided between the lower board part, the upper board part joined to the surface of the said lower board part, and the said lower board part and the said upper board part. And a lower temperature measuring element for measuring the temperature of a lower external contact of the cavity, which is used for a chemical reaction of a reactant contained in the cavity by heating and cooling from the outside. And a chemical reaction chip configured to measure the temperature of the upper external contact of the cavity in the vertical direction from the lower temperature measuring element of the upper plate part, Perform temperature control that repeats heating and cooling, measure the temperature of the lower part of the chip for chemical reaction , measure the temperature of the upper part of the chip for chemical reaction, and connect to the lower temperature measuring element and the upper temperature measuring element And When estimating the temperature of the reactant stored in the chemical reaction chip using the degree estimation algorithm, the temperature estimation algorithm is used to obtain the measurement results obtained from the lower temperature measurement element and the upper temperature measurement element. Time series data when the heating and cooling from the outside to the cavity of the chemical reaction chip are repeated to obtain frequency components, and the frequency components are used for the chemical reaction. A temperature control method characterized by estimating a time-series temperature inside a chip .

(5) Moreover, another one aspect | mode of this invention is the computer of a temperature control apparatus, a lower board part, the upper board part joined to the surface of the said lower board part, the said lower board part, and the said upper board part A chip for chemical reaction used for a chemical reaction of a reactant contained in the cavity by heating and cooling from the outside, and a temperature of a lower external contact of the cavity Accommodated in a chip for chemical reaction composed of a lower temperature measuring element for measuring the temperature and an upper temperature measuring element for measuring the temperature of the upper external contact of the cavity in the vertical direction from the lower temperature measuring element of the upper plate portion Temperature control means for performing temperature control for repeating the heating and cooling of the reaction product, and connected to the lower temperature measuring element and the upper temperature measuring element, and for the chemical reaction using a temperature estimation algorithm Functioning as temperature estimation means for estimating the temperature of the reactant contained in the cup, the temperature estimation means using a temperature estimation algorithm, the measurement obtained from the lower temperature measurement element and the upper temperature measurement element Time series data of the results, each of the time series data when repeating the heating and cooling from the outside to the cavity of the chemical reaction chip to obtain a frequency component, using the frequency component It is a program characterized by estimating the time series temperature inside the chemical reaction chip.

  According to the present invention, it is possible to estimate the internal temperature of a measurement object containing a small and brittle substance from the external temperature in a non-contact manner.

FIG. 1A is an exploded perspective view showing the configuration of a chemical reaction chip according to an embodiment of the present invention. FIG. 1B is a side view showing the configuration of the chemical reaction chip. FIG. 2 is an enlarged cross-sectional view of the chemical reaction chip. FIG. 3 is a functional block diagram of the temperature control device. FIG. 4 is a diagram illustrating a processing flow of the temperature control device. FIG. 5 is a diagram showing the transition of the measured temperature of the upper external contact A. As shown in FIG. FIG. 6 is a diagram showing the transition of the measured temperature of the lower external contact C. FIG. 7 is a graph showing the measured temperature of the upper external contact A, the measured temperature of the lower external contact C, and the estimated temperature at point B in the aqueous solution.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 ... Upper board part 2 ... Lower board part 3 ... Aqueous solution injection hole 4 ... Recessed part 5 ... Upper temperature measurement point (upper temperature measurement element)
6 ... Lower temperature measurement point (lower temperature measurement element)
DESCRIPTION OF SYMBOLS 10 ... Temperature control apparatus 11 ... Input-output part 12 ... Control part 13 ... Internal temperature estimation part 14 ... Temperature control part 15 ... Memory | storage part

Hereinafter, a chemical reaction chip and a temperature control device according to an embodiment of the present invention will be described with reference to the drawings.
In addition, this invention is applicable to the use which changes the temperature of the chip | tip for chemical reaction repeatedly, such as PCR method. Further, the present invention can also be suitably applied to an application for maintaining the temperature of the chemical reaction chip at a predetermined constant temperature.
1A and 1B are diagrams showing a configuration of a chemical reaction chip according to the embodiment.
1A is an exploded perspective view of a chip for chemical reaction, and FIG. 1B is a cross-sectional view of the chip for chemical reaction. In these drawings, an upper plate portion 1 (including polypropylene), a lower plate portion 2 (including aluminum), an aqueous solution injection hole 3, a recess 4, an upper temperature measuring element 5, and a lower temperature measuring element 6 are illustrated. ing. As shown in FIG. 1A, in the chip for chemical reaction, corresponding surfaces of the upper plate portion 1 and the lower plate portion 2 are joined. The upper plate part 1 is provided with two injection holes 3 for the DNA aqueous solution. The lower plate 2 is provided with a recess 4 in which an aqueous solution is accumulated. As shown in the cross-sectional view of FIG. 1B, a lower temperature measuring element 6 (for example, a thermistor, a Peltier element, etc.) that measures the temperature of this contact is provided at the lower end (lower external contact C) of the recess 4 of the lower plate part 2. Is provided. Further, an upper temperature measuring element 5 (for example, thermistor, Peltier) for measuring the temperature of this contact is provided at the upper end (upper external contact A) at a position along the vertical direction from the lower temperature measuring element 6 on the upper surface of the upper plate 1. Element). The upper temperature measurement element 5 and the lower temperature measurement element 6 are signal-connected to a temperature control device that receives input of measurement values of the elements. 1A and 1B show a configuration in which the lower plate portion 2 has a recess, but a configuration in which the upper plate portion 1 is provided with a recess and the structures of the upper plate portion 1 and the lower plate portion 2 are interchanged. It may be used. That is, any structure having a cavity (space) in which a DNA aqueous solution is stored in a flat plate may be used.

FIG. 2 is an enlarged sectional view of the chemical reaction chip.
In this figure, A is an upper external contact measured by the upper temperature measuring element 5 shown in FIGS. 1A and 1B. C is a lower external contact measured by the lower temperature measuring element 6 shown in FIGS. 1A and 1B. The temperature control device estimates the temperature at the temperature estimation point B inside the aqueous solution. In the present embodiment, the upper plate portion 1 is made of polypropylene and the lower plate portion 2 is made of aluminum, but a combination of other materials may be used. As shown in FIG. 2, the thickness of the upper plate portion 1 is d1, and the thickness of the lower plate portion 2 is d4. The point B in the aqueous solution of the recess 4 is located at a distance of d2 from the upper plate portion 1 and d3 from the lower plate portion. In the steady state, the temperatures at points A, B, and C are constant regardless of time. The temperature TB of the aqueous solution is intermediate between the temperature TA at the point A of the upper plate portion 1 and the temperature TC at the point C of the lower plate portion 2.

FIG. 3 is a functional block diagram of the temperature control device.
As shown in this figure, the temperature control device 10 includes an input / output unit 11, a control unit 12, an internal temperature estimation unit 13, a temperature control unit 14, and a storage unit 15. Here, the input / output unit 11 receives an input of the measured temperature from the upper temperature measuring element 5 or the lower temperature measuring element 6 of the chemical reaction chip. Further, the control unit 12 controls each processing unit of the temperature control device 10. The internal temperature estimation unit 13 estimates the temperature inside the aqueous solution accumulated in the recess of the chemical reaction chip. Further, the temperature control unit 14 (for example, a heater, a heat sink, a Peltier element, etc.) performs feedback control based on a temperature estimation value that is a result of temperature estimation by the internal temperature estimation unit 13 and a desired temperature inside the aqueous solution recorded in advance. To control the temperature of heating or cooling to bring the estimated temperature close to the desired temperature.
The storage unit 15 also stores parameters for estimating the temperature inside the aqueous solution using a temperature estimation algorithm, parameters for controlling the internal temperature of the aqueous solution using feedback control, and the like.

FIG. 4 is a diagram showing a processing flow of the temperature control device.
Next, the processing flow of the temperature control apparatus 10 will be described. Note that the temperature estimation method of the temperature control device 10 in the present embodiment ignores the convection of the aqueous solution in the recess 4 and regards the upper plate portion 1, the aqueous solution, and the lower plate portion 2 as heat conductors of the layered material. .
First, periodic heating / cooling is repeatedly performed on the chemical reaction chip in which an aqueous solution containing DNA is injected into the recess 4 based on the PCR method. At this time, the internal temperature estimation unit 13 of the temperature control device 10 transmits the measurement temperature at the upper external contact A from the upper temperature measurement element 5 and the lower external from the lower temperature measurement element 6 via the input / output unit 11. The input of the measured temperature of the contact C is received.

FIG. 5 is a diagram showing the transition of the measured temperature of the upper external contact A. FIG.
FIG. 6 is a diagram showing the transition of the measured temperature of the lower external contact C. In FIG.
As shown in FIGS. 5 and 6, the internal temperature estimation unit 13 acquires the measured temperature of the upper external contact A and the measured temperature of the lower external contact C from the upper temperature measuring element 5 and the lower temperature measuring element 6. The storage unit 15 stores the acquired data. The measured temperature shown in FIGS. 5 and 6 is measured data corresponding to one cycle [T] when heating or cooling of the chemical reaction chip is repeated.

  Here, since there is no heat source inside the cavity constituted by the recesses 4 and the like, the heat conduction equation in each material is expressed by equation (1). Here, ρ represents density, C represents specific heat, k represents a heat transfer coefficient, and u (t, x) represents temperature. Further, the density ρ, the specific heat C, and the heat conduction coefficient k can be expressed by Expression (2).

In the formula (2), M represents mass, L represents length, and T represents time. Considering temperature measurement data (FIGS. 5 and 6) in one cycle [T] when repeating heating or cooling of the chemical reaction chip in the PCR method, the Fourier component U _n (x) is expressed by the following equation (3). Can be represented by

  Here, the angular velocity in one cycle [T] when heating or cooling the chemical reaction chip is repeated is as follows:

  Therefore, the temperature u (t, x) can be expressed by the equation (5).

Further, when the formula (5) is substituted into the formula (1), the following formula is obtained.

Here, the following W _n is introduced. W _n corresponds to a spatial change amount of the temperature U and represents a temperature gradient adjusted in dimension.

  In this case, Equation (6) can be expressed as follows.

Therefore, when the solutions of the equations (7) and (8) are obtained, the following equations are obtained.

Here, the following formula is used.

Further, the following formula is established.

  Now, it is assumed that the temperature is the same at the interface between the upper plate portion 1 and the aqueous solution, and the heat flow entering the interface and the heat flow exiting are the same. That is, when x = d1 (corresponding to the thickness of the upper plate portion 1), it can be expressed as follows.

  Moreover, when rewritten using Formula (13), it can represent as follows.

  The boundary condition that is a combination of Expression (12) and Expression (14) can be expressed by the following expression.

  The following formula is used.

  Moreover, it can represent as follows by Formula (9) and Formula (15).

Here, the following equation is a transfer matrix that connects x = d1 and x = d2.

  The following formulas are the same as above.

The above formula (22) is used. And the following formula is used.

T ₃₂ is a transfer matrix connecting x = d2 and x = d3, T ₄₃ is a transfer matrix connecting x = d3 and x = d4, T ₃₁ is a transfer matrix connecting x = d1 to x = d3, T Reference numeral ₄₁ denotes a transfer matrix that connects x = d1 to x = d4.

  S shown in the above equation (25) (transfer matrix linking temperature and temperature gradient (U and W) at points A and C) is introduced. As a result, the following relationship is derived.

  Then, when equation (26) is solved for W, the following equation is obtained.

  Using the equation (28), the following equation is obtained.

Point B in the aqueous solution by the equation (29), that is, the temperature ³ U _n (xi] ₂₎ in the x = d2 is obtained. In the above equation, ξ ₁ , ξ ₂ , ξ ₃ , and ξ ₄ are obtained by making the distances x = d1, d2, d3, and d4 dimensionless.

That is, in the method for calculating the temperature of the point B in the aqueous solution described using the above equations (1) to (29), first, the internal temperature estimation unit 13 of the temperature control device 10 is connected via the input / output unit 11. Then, the measurement temperature of the upper external contact A is received from the upper temperature measurement element 5, and the measurement temperature of the lower external contact C is received from the lower temperature measurement element 6 (step S1). The Fourier transform of time series data for each of its temperature, the frequency components determined (step S2), and this ¹ U _n ^(0), and _{4 U n} (ξ 4). Further, for each frequency component (n), ξ ₁ , ξ ₂ , ξ ₃ , ξ ₄ shown above are calculated (step S3), and T ₄₃ , T ₃₂ , T ₂₁ are calculated (step S3). S4). _Also, determine the _{T 31 = T 32 T 21,} T 41 = T 43 T 31 ( step S5).

  Further, the above equation (30) is obtained (step S6).

Further, the above equation (31) Temperature ³ Request U _n (xi] ₂₎ (step S7). Further, the time series temperature ³ u (t, d ₂ ) at the point B in the aqueous solution is obtained by performing inverse Fourier transform on the sequence of ³ U _n (ξ ₂ ) obtained for each frequency component (n) in this way. Is calculated (step S8).

  FIG. 7 is a graph showing the measured temperature of the upper external contact A, the measured temperature of the lower external contact C, and the estimated temperature at point B in the aqueous solution. According to the present embodiment, as shown in FIG. 7, it is possible to estimate the temperature without contact with the temperature measurement object (point B in the aqueous solution in the present embodiment).

  The above temperature control apparatus has a computer system inside. Each process described above is stored in a computer-readable recording medium in the form of a program, and the above process is performed by the computer reading and executing the program. Here, the computer-readable recording medium means a magnetic disk, a magneto-optical disk, a CD-ROM, a DVD-ROM, a semiconductor memory, or the like. Alternatively, the computer program may be distributed to the computer via a communication line, and the computer that has received the distribution may execute the program.

The program may be for realizing a part of the functions described above.
Furthermore, what can implement | achieve the function mentioned above in combination with the program already recorded on the computer system, and what is called a difference file (difference program) may be sufficient.

  According to the present invention, it is possible to estimate the internal temperature of a measurement object containing a small and brittle substance from the external temperature in a non-contact manner.

Claims (5)

A lower plate portion, an upper plate portion joined to a surface of the lower plate portion, and a cavity provided between the lower plate portion and the upper plate portion, and by heating and cooling from the outside, A chemical reaction chip used for a chemical reaction of a reactant contained in the cavity, the lower temperature measuring element for measuring the temperature of the lower external contact of the cavity, and the vertical temperature from the lower temperature measuring element of the upper plate portion A temperature control unit that performs temperature control that repeats the heating and cooling of the reactant contained in a chemical reaction chip configured with an upper temperature measuring element that measures the temperature of the upper external contact of the cavity in the direction ; ,
A temperature estimation unit that is connected to the lower temperature measurement element and the upper temperature measurement element and performs temperature estimation of a reactant contained in the chemical reaction chip using a temperature estimation algorithm ;
Bei to give a,
The temperature estimation unit is a time-series data of measurement results obtained from the lower temperature measurement element and the upper temperature measurement element using a temperature estimation algorithm, from the outside to the cavity of the chemical reaction chip The time series data when repeating heating and cooling are respectively Fourier transformed to obtain frequency components, and the time series temperature inside the chemical reaction chip is estimated using the frequency components.
A temperature control device characterized by that . The temperature estimation unit is a time-series data of measurement results obtained from the lower temperature measurement element and the upper temperature measurement element using a temperature estimation algorithm, from the outside to the cavity of the chemical reaction chip Each time-series data when repeating heating and cooling is Fourier transformed to obtain frequency components,
A thickness d1 of the upper plate portion on a straight line connecting the upper external contact and the lower external contact, a thickness obtained by subtracting the thickness d1 from a distance between a predetermined temperature estimation point in the cavity and the upper external contact d2, thickness d4 of the lower plate portion, and thickness d1 and thickness d2 when the thickness d4 is obtained by subtracting the thickness d4 of the plate portion from the distance between the temperature estimation point and the lower external contact. , The temperature transfer matrix connecting the thickness d2 and the thickness d3, the temperature transfer matrix connecting the thickness d3 and the thickness d4, and the temperature connecting the thickness d1 to the thickness d3. Obtained by Fourier transform, respectively, the transfer matrix of temperature, the transfer matrix of temperature connecting the thickness d1 to the thickness d4, and the time series data of the measurement results obtained from the lower temperature measurement element and the upper temperature measurement element And the frequency component to estimate the temperature of the temperature estimation point. Make
The temperature control apparatus according to claim 1. Temperature control for performing temperature control of heating and cooling so that the temperature estimated value approaches the desired temperature based on the temperature estimated value obtained by the temperature estimating unit and the desired temperature of the reactant recorded in advance. temperature control device according to claim 1 or claim 2, further comprising a part. A lower plate portion, an upper plate portion joined to a surface of the lower plate portion, and a cavity provided between the lower plate portion and the upper plate portion, and by heating and cooling from the outside, A chemical reaction chip used for a chemical reaction of a reactant contained in the cavity, the lower temperature measuring element for measuring the temperature of the lower external contact of the cavity, and the vertical temperature from the lower temperature measuring element of the upper plate portion Performing temperature control for repeating the heating and cooling of the reactant contained in a chemical reaction chip configured with an upper temperature measurement element that measures the temperature of the upper external contact of the cavity in the direction,
Measure the temperature at the bottom of the chemical reaction chip,
Measure the temperature of the top of the chemical reaction chip,
The temperature measurement algorithm is used to estimate the temperature of the reaction product connected to the lower temperature measurement element and the upper temperature measurement element and stored in the chemical reaction chip using the temperature estimation algorithm. Time series data of measurement results obtained from the element and the upper temperature measurement element, each time series data when repeating heating and cooling from the outside to the cavity of the chemical reaction chip, respectively, Fourier transform A frequency component is obtained, and the time-series temperature inside the chemical reaction chip is estimated using the frequency component.
A temperature control method characterized by that . The temperature control computer,
A lower plate portion, an upper plate portion joined to a surface of the lower plate portion, and a cavity provided between the lower plate portion and the upper plate portion, and by heating and cooling from the outside, A chemical reaction chip used for a chemical reaction of a reactant contained in the cavity, the lower temperature measuring element for measuring the temperature of the lower external contact of the cavity, and the vertical temperature from the lower temperature measuring element of the upper plate portion A temperature control means for performing temperature control to repeat the heating and cooling of the reactant contained in a chemical reaction chip configured with an upper temperature measuring element that measures the temperature of the upper external contact of the cavity in the direction;
A temperature estimation means connected to the lower temperature measurement element and the upper temperature measurement element, for estimating a temperature of a reactant contained in the chemical reaction chip using a temperature estimation algorithm;
Function as
The temperature estimation means is time-series data of measurement results obtained from the lower temperature measurement element and the upper temperature measurement element using a temperature estimation algorithm, from the outside to the cavity of the chemical reaction chip The time series data when repeating heating and cooling are respectively Fourier transformed to obtain frequency components, and the time series temperature inside the chemical reaction chip is estimated using the frequency components.
A program characterized by that.