WO2005068066A1 - Temperature controller and protein crystallizer utilizing the same - Google Patents

Abstract

A temperature controller that by creating of a temperature gradient on a heat conductor surface, can simultaneously adjust multiple samples disposed on the surface to different temperatures and can regulate a temperature profile of the heat conductor surface as desired. There is provided a temperature controller comprising a heating/cooling device and a heat conductor disposed in thermal contact with the heating/cooling device, wherein in accordance with the temperature of the heat conductor, the temperature of samples disposed in thermal contact therewith is regulated. In the temperature controller, two or more heating/cooling devices are disposed, and by setting of these for different temperatures, a temperature gradient is created on a surface of heat conductor in thermal contact with the samples. The heat conductor is composed of two or more types of materials whose values dc/K, wherein the specific gravity is referred to as d[kg/m3], the specific heat as c[J/(kg·K)] and the thermal conductivity as K[W/(m·K)], are different from each other.

Description

 Temperature control device and protein crystallization device using the same

 Technical field

 The present invention relates to a temperature control device used for temperature control of a sample, and more particularly, to a temperature control device capable of simultaneously adjusting a plurality of samples to different temperatures. Background art

 [0002] In the field of chemistry, various experimental operations such as synthesis of chemical substances and evaluation of their properties have been performed. An example of such an experimental operation is a protein crystallization operation. The crystallization of the protein is mainly performed to analyze the three-dimensional structure of the protein by X-ray crystal structure analysis. Information on the three-dimensional structure of a protein is essential for understanding its specific properties and functions. For example, in the basic chemistry field, it becomes the basis for understanding the mechanism of function expression in biochemical systems by enzymes and hormones, and in the industrial field, it is used to develop new drugs based on protein function analysis. Be utilized.

 [0003] In such various experimental operations, temperature is one of the most important conditions. Therefore, in order to find the optimal temperature condition, the experimental operation is repeated under various temperature conditions, that is, temperature screening is performed. In particular, protein crystallization is very sensitive to external conditions such as temperature, and setting such conditions is often difficult. Therefore, temperature screening is a very important operation. A thermostat is usually used to control the temperature of the experimental system. However, in a conventional thermostatic chamber, the temperature inside the chamber was maintained at a uniform temperature, so that only one experiment operation could acquire experimental data under a certain temperature condition. Therefore, in order to acquire experimental data under various temperature conditions, after completing the experiment under one temperature condition, reset the temperature of the constant temperature bath to another temperature and wait until the set temperature is stably maintained. After that, it is necessary to repeat the operation of performing the next experiment, and there is a problem that much time and labor are required.

[0004] As a solution to such a problem, there has been proposed a constant temperature measuring device with a temperature gradient, which enables measurement under a plurality of temperature conditions at the same time (Patent Document 1). With this temperature gradient The constant temperature measuring device has a configuration in which a heat generating mechanism is provided at one end of a heat conductor made of brass, and a heat radiating mechanism is provided at the other end of the heat conductor. In addition, a plurality of temperature sensors are provided to measure the temperature at each point on the heat conductor surface. In this measuring instrument, the heat generated by the heat generating mechanism is transmitted through the heat conductor, and is radiated to the heat radiating mechanism. As a result, a temperature gradient from the heat generating mechanism side to the heat radiating mechanism side is formed on the heat conductor surface, and the temperature of the heat conductive surface varies depending on the position. By placing samples at each position on the heat conductor surface, it is possible to adjust a plurality of samples simultaneously to different temperatures.

 Patent Document 1: JP-A-7-318522

 Disclosure of the invention

 Problems to be solved by the invention

[0005] However, there has been a problem that it is very difficult to adjust the temperature profile file on the surface of the heat conductor as desired with the above-mentioned constant temperature measuring device with a temperature gradient. In particular, when a relatively small temperature difference (e.g., about 120 ° C in number) occurs near or below room temperature in a heat conductor, the heat dissipation or heat absorption generated between the heat-generating part and the heat-absorbing part Since the influence on the temperature profile of the heat conductor becomes large, it has been very difficult to realize a desired temperature profile. Therefore, for example, when the arrangement of samples is fixed, such as in an experiment using a multi-hole plate, the temperature of each sample may not always be able to be set to a desired temperature.

[0006] Therefore, the present invention provides a method of forming a temperature gradient on the surface of a heat conductor so that a plurality of samples placed on the surface can be easily and simultaneously adjusted to different temperatures. An object of the present invention is to provide a temperature control device capable of adjusting a surface temperature profile as desired, and a protein crystallization device using the same.

 Means for solving the problem

[0007] In order to solve the above-mentioned problems, a temperature control device of the present invention includes a heating / cooling element, and a heat conductor arranged in thermal contact with the heating / cooling element. A temperature adjusting device for adjusting the temperature of a sample placed in thermal contact with the temperature according to the temperature of the sample, wherein the heating and cooling elements are provided in a number of two or more, and these heating and cooling elements are Mutual When the temperature is set to a different temperature, a temperature gradient is formed on the surface of the heat conductor that is in thermal contact with the sample, and the specific gravity of the heat conductor is d [kgZm ³ ]. , Specific heat. ^! /! ^ '! ^)], And the thermal conductivity is K [WZ (m'K)], the value represented by dcZK must be composed of two or more different materials. It is characterized by.

 [0008] Further, in the temperature control device of the present invention, the heat conductor is replaced with two or more materials having different dcZK values from each other, but instead of the heat conductor. Alternatively, a plurality of recesses may be formed on the surface opposite to the surface that is in thermal contact with the sample, the air being present in the recesses.

 [0009] Further, the protein crystallization apparatus of the present invention is an apparatus for precipitating the protein crystals from a sample solution containing a protein, wherein the temperature of the sample solution is adjusted to adjust the temperature of the sample solution. It is characterized by using an adjusting device.

 The invention's effect

 [0010] According to the temperature control device of the present invention, as described above, by setting the plurality of heating / cooling elements to different temperatures, the temperature gradient is applied to the sample placement surface of the heat conductor in thermal contact therewith. An arrangement is formed. That is, the temperature of the heat conductor is maintained at different positions on the sample placement surface depending on the position. Therefore, by placing a plurality of samples at different positions on the heat conductor, these samples can be simultaneously adjusted to different temperatures.

 [0011] Further, in the temperature control device of the present invention, the heat conductor is made of two or more kinds of materials having different dcZK values, or the heat conductor is provided on the surface of the heat conductor opposite to the sample placement surface. A plurality of concave portions (there is air therein) are formed. Therefore, the temperature profile on the sample placement surface of the heat conductor can be easily adjusted according to the purpose of use of the device by selecting the combination of the materials constituting the heat conductor or the shape of the recess. It is possible.

 [0012] Further, the protein crystallization apparatus of the present invention uses the above-described temperature control apparatus of the present invention, and therefore can simultaneously perform crystallization under a plurality of temperature conditions. This has the advantage that temperature screening can be performed efficiently.

Brief Description of Drawings FIG. 1 is a cross-sectional view showing one example of a temperature control device of the present invention.

 FIG. 2 is a cross-sectional view showing an example of a heat conductor constituting the above-mentioned temperature control device.

FIG. 3 is a cross-sectional view showing another example of a heat conductor constituting the above-mentioned temperature controller.

 FIG. 4 is a cross-sectional view showing still another example of the heat conductor constituting the temperature control device.

 FIG. 5 is a cross-sectional view showing still another example of the heat conductor constituting the temperature control device.

 [FIG. 6] FIG. 6 is a cross-sectional view showing still another example of the heat conductor constituting the above-mentioned temperature control device.

 [FIG. 7] FIG. 7 is a cross-sectional view showing still another example of the heat conductor constituting the temperature control device.

 FIG. 8 is a cross-sectional view showing still another example of the heat conductor constituting the temperature control device.

 FIG. 9 is a cross-sectional view showing one example of the protein crystallization apparatus of the present invention.

 FIG. 10 is a cross-sectional view showing another example of the protein crystallization apparatus of the present invention. A is the cross-sectional view, and B is a cross-sectional view in which the cross-sectional direction of A and the cutting direction are orthogonal to each other.

 FIG. 11 is a cross-sectional view showing a configuration of a heat conductor used in Example 1. A is a cross-sectional view showing the configuration of the heat conductor used in Example 11; B is a cross-sectional view showing the configuration of the heat conductor used in Example 12; and C is a comparative example. FIG. 3 is a cross-sectional view showing a configuration of a heat conductor used.

 FIG. 12 is a graph showing the relationship between the position of the heat conductor surface and the temperature gradient in the temperature control device manufactured in Example 1.

 FIG. 13 is a graph showing the relationship between the arrangement position of samples and the adjusted sample temperature in the temperature adjustment using the temperature adjustment device manufactured in Example 1.

FIG. 14 is an optical micrograph of a protein crystal formed in Example 2. FIG. 15 is a graph showing the relationship between the position of the heat conductor surface and the temperature in the temperature controller manufactured in Example 3. A is the graph when the set temperatures of the two Peltier elements are set to 5 ° C and 15 ° C, respectively, and B is the set temperature of the two Peltier elements at 5 ° C and 50 ° C, respectively. 5 is the graph when the temperature is set to ° C.

 FIG. 16 is a cross-sectional view showing a configuration of another heat conductor used in Example 1.

 FIG. 17 is a diagram showing the relationship between the control set temperature of the control device and the temperature generated on the heat conductor in the temperature control device manufactured in Example 1. A is a graph showing the relationship between the set average temperature of the control device and the actually measured average temperature of the heat conductor, and B is a graph showing the relationship between the set temperature difference of the control device and the generated temperature gradient. .

 FIG. 18 is a plan view showing an example of a method of embedding a temperature measuring element in a heat conductor according to the present invention.

 FIG. 19 is a plan view showing another example of a method of embedding a temperature measuring element in a heat conductor according to the present invention.

 FIG. 20 is a cross-sectional view showing an example of an arrangement of a temperature gradient generation unit and a control device in the temperature control device of the present invention.

 FIG. 21 is a cross-sectional view showing another example of the temperature control device of the present invention.

 FIG. 22 is a cross-sectional view showing an example of the installation of another temperature measuring element in the temperature control device.

 FIG. 23 is a schematic view showing one example of a part of the temperature control device of the present invention.

 FIG. 24 is a graph showing an example of temperature control by the above-mentioned temperature controller. A is

Is a graph showing an example of gradually increasing the temperature gradient at a constant temperature, B is a graph showing an example in which the temperature is lowered as a whole while maintaining a constant temperature gradient, and C is a graph showing the average temperature. It is a graph which shows the example which changes a temperature gradient gradually without changing.

 FIG. 25 is a diagram showing an example of another heat conductor constituting the temperature control device and an example of disposition of the another heat conductor on the heat conductor. A is a cross-sectional view of the another heat conductor, and B is a plan view showing an example of the arrangement of the another heat conductor on the heat conductor.

FIG. 26 is a graph showing the position of the heat conductor surface in the temperature controller manufactured in Example 4. 6 is a graph showing the relationship between the position and the temperature in the well.

 FIG. 27 is a cross-sectional view showing another example of another heat conductor constituting the above-mentioned temperature control device.

 FIG. 28 is an example of the arrangement of the another heat conductor and the second another heat conductor on the heat conductor, and the another heat conductor and the second another heat conductor. FIG. 4 is a diagram showing the relationship between the position of the surface of the sphere and the temperature. A is a plan view of an example of the arrangement of the another heat conductor and the second another heat conductor on the heat conductor, B is the another heat conductor and the second heat conductor 7 is a graph showing the relationship between the position of the surface of another heat conductor and the temperature.

FIG. 29 is a schematic view showing another example of a part of the temperature control device of the present invention. Explanation of symbols

 la, lb, 14 heating / cooling element

 2, 12 Thermal conductor

 3 Heat bath

 4 plates

 5 samples

 6, 18 Thermometer

 7 Insulation cover

 8 grooves

 9 recess

 10 housing

 11 Holding mechanism

 13 Insulation

 15 Aluminum parts

 16 SUS member

 17 Yet another heat conductor

 19 Thermometer insertion hole

 20 Temperature sensor

21 Temperature gradient generation unit 22 Power

 23 Control board

 24 transparent acrylic board

 25 air layer

 26 Lid

 27 O-ring

 28 Thermometer holder

 29, 34 Layer with large dcZK value

 30, 33 Low dcZK value layer

 32, 36 different thermal conductors

 37 Different temperature sensors

 BEST MODE FOR CARRYING OUT THE INVENTION

 [0015] As described above, the temperature control device of the present invention is provided with a heat conductor in thermal contact with a plurality of heating / cooling elements set at different temperatures, and a temperature gradient is applied to the surface of the heat conductor. Is formed.

 [0016] Suppose that the heat conductor is composed of a single material of a single material (that is, the dcZK value is kept constant throughout the entire body), and two heat sources set at different temperatures are used. When the cooling elements are brought into contact, a temperature gradient is formed in a region corresponding to between the heating and cooling elements. However, at this time, the magnitude of the temperature gradient is not uniform in the entire area, and becomes small at both ends of the area which is large at the center of the area. In addition, comparing the high temperature side end with the low temperature side end, the temperature gradient force is smaller at the low temperature side end. Such a tendency of the temperature profile cannot be substantially changed even if the set temperature of the heating / cooling element is changed, as long as one kind of material is used as the heat conductor.

[0017] Therefore, in the present invention, the value obtained by dividing the heat capacity per unit volume of the heat conductor by the heat conductivity, that is, the specific gravity is d [kgZm ³ ], and the specific heat is calculated by the following formula: ! ^, Where the thermal conductivity is! ^ ¹ ^ Z (mK)] The force expressed by dcZK [sZm ² ] By using two or more different materials, a desired temperature profile is achieved. ing.

[0018] The dcZK value indicates how easily the internal temperature of the material becomes uniform and how easily a temperature gradient occurs. It is an index. A material having a small dcZK value has a property that its internal temperature is easily made uniform. In other words, a material having a small dcZK value tends to be small even if it is placed in a non-uniform temperature environment, or if it is difficult to form a temperature gradient. On the other hand, a material having a large dcZK value has a property that its internal temperature is difficult to be uniform. That is, a material having a large dcZK value tends to form a large temperature gradient when placed in a non-uniform temperature environment.

 In the temperature control device of the present invention, a desired temperature profile is realized by combining a plurality of materials having different dcZK values while considering the relationship between the dcZK value and the temperature gradient as described above. Can be.

 For example, a desired temperature profile force is applied to the surface of the heat conductor in contact with the sample so that the temperature gradient is substantially uniform (in other words, the distance from one end of the temperature gradient forming region (temperature gradient). If the temperature increases or decreases linearly with an increase in the distance in the arrangement direction), the following configuration can be adopted. The heating and cooling elements are arranged on the lower surface of the heat conductor and the sample is arranged on the upper surface.The upper and lower surfaces of the heat conductor are made of materials with different dcZK values, and the dcZK value of the material forming the upper surface is set. Is adjusted to be smaller than the dcZK value of the material constituting the lower surface. According to such a configuration, a large temperature gradient is formed on the surface in contact with the heating / cooling element, while the temperature gradient is reduced on the surface in contact with the sample, that is, a smooth temperature profile is formed. While ensuring the difference, it is possible to realize a uniform temperature gradient on the surface in contact with the sample.

 Further, as described above, the tendency of the temperature profile formed when the thermal conductor is made of a material having a constant dcZK value and the deviation from the desired temperature profile are recognized, and the ZK value and the temperature are determined. By adjusting the dcZK value distribution of the heat conductor by combining multiple materials with different dcZK values so as to correct this shift while taking into account the relationship with the gradient, the desired temperature profile can be adjusted more precisely. Can be realized.

[0022] When considering the dcZK value distribution of the heat conductor, if the average value of the dcZK value is used as the dcZK value of the heat conductor, it becomes easier to adjust the dcZK value distribution of the heat conductor. This average value is calculated by setting the dcZK value of each material constituting the heat conductor to (dcZK) x and the temperature gradient of the heat conductor. When the area occupied by each material in a cross section perpendicular to the formation direction is Sx [m ² ], the value is represented by the following formula.

[0023] [Equation 1]

∑ {(d c / K) x · S x} / ^ χ

Here, x is an integer of η. Η is the number of materials constituting the heat conductor and is an integer of 2 or more, for example, an integer in the range of 2-8, preferably 2-4.

 [0024] For example, when the desired temperature profile force is substantially uniform in a region where the temperature gradient of the heat conductor is formed, if the temperature gradient is substantially uniform in the region where the temperature gradient of the heat conductor is formed, dcZK What is necessary is just to adjust the average value so that it is smaller at the center of the region than at the end of the region. Furthermore, the dcZK value (average value) distribution in the direction of forming the temperature gradient of the heat conductor may be adjusted so as to gradually increase as the force moves toward the center of the force. With such a configuration, the difference in temperature gradient between the central portion and both ends of the heat conductor can be reduced, and the relationship between the dcZK value and the temperature gradient as described above can be understood.

 [0025] Further, in order to improve the uniformity of the temperature gradient, in the region where the temperature gradient is formed, the partial force at which the average value of the dcZK value of the heat conductor is minimized is higher than that in the center of the region. It is preferably located on the high temperature side. With such a configuration, as described above, the difference in temperature gradient between the high-temperature side end and the low-temperature side end can be reduced, and a further uniform temperature gradient can be realized. is there.

 [0026] Further, in a region where the temperature gradient of the heat conductor is formed, the temperature gradient is made non-uniform, and the temperature increases or decreases stepwise as the distance of the one-end force in the region increases. If such a temperature profile is desired, the average value of the dc / K value in the temperature gradient forming region of the heat conductor may be adjusted so as to be larger at the center of the region than at the end of the region. . With such a configuration, the temperature gradient at the central portion of the heat conductor can be further increased, and a mode in which the temperature changes stepwise at the central portion can be realized.

[0027] Furthermore, in the temperature gradient forming region of the heat conductor, the average value of the dcZK value is large, If small portions are provided and arranged alternately along the temperature gradient forming direction, it is possible to realize a form in which the temperature changes in multiple stages.

In the case of a deviation, the average value of the dcZK value is 8 × 10 ⁶ s / m at each position of the heat conductor so that an excessive heat resistance does not occur in the heat conductor. It is preferable to set it to ² or less, and it is preferable to set it to 1 × 10 ³ sZm ² or more at each position of the heat conductor so that excessive temperature uniformity does not occur. In addition, the difference between the maximum and minimum parts of the average value of the dcZK value is set as appropriate according to the distance between the heating and cooling elements, the difference in the set temperature, etc., which is not particularly limited. The force that can be applied, for example 5 × 10 ³ s / m ² —5 × 10 ⁵ s / m ² , preferably 2 × 10 ⁴ sZm ² —3 × 10 ⁵ sZm ² .

 There are no particular restrictions on the material constituting the heat conductor as long as it is at least two, but for example, 2-8, preferably 2-4.

 [0030] Instead of configuring the thermal conductor with two or more materials having different dcZK values, the thermal conductor is composed of one or more materials and is opposite to a surface that is in thermal contact with the sample. A desired temperature profile can also be realized by forming a plurality of recesses on the surface and allowing air to exist in the recesses.

 [0031] Air has a dcZK value that is larger than that of the metal or the like that forms the heat conductor. By providing a portion containing air in the heat conductor, the heat conductor can be separated into two types having different dcZK values. The same effects as those made of the above materials can be realized.

 In this case, the average value of the dcZK value is determined using the above equation, based on the assumption that the concave portion is filled with a solid substance (virtual substance) having the same dcZK value as air. be able to.

 Hereinafter, embodiments of the present invention will be described with reference to the drawings. In each of the embodiments described below, the temperature gradient becomes substantially uniform over the entire temperature gradient forming region (the temperature increases or decreases linearly with an increase in the distance of one end force in the region). This is an example of a case where such a temperature profile is realized. However, as described above, the present invention can be applied to realize various temperature profiles which are not limited thereto.

(First Embodiment) FIG. 1 is a configuration diagram showing an example of the temperature control device according to the first embodiment of the present invention. This temperature control device includes a plurality of heating / cooling elements la and lb, and a heat conductor 2 arranged in thermal contact with the upper surface thereof. Further, on the lower surface of the heating and cooling elements la and lb (the surface opposite to the surface in contact with the heat conductor 2), a heat bath section 3 for recovering exhaust heat is arranged. In order to suppress heat radiation or heat absorption from the heating / cooling elements la and lb and the heat conductor 2, a heat insulating cover 7 made of, for example, a fluorine resin is disposed so as to cover these side surfaces. Further, the present apparatus is provided with a control device (not shown) for driving and controlling the heating and cooling elements la and lb. The control device includes, for example, a power supply, a control circuit, and a control board on which the control circuit is mounted. The configuration shown in FIG. 1 excluding the control device may be hereinafter referred to as a temperature gradient generation unit.

 The heating / cooling elements la and lb are heat sources for heating or cooling the heat conductor 2, and a plurality of heating / cooling elements la and lb are provided for one heat conductor 2. In FIG. 1, the case where two heating / cooling elements are used will be described as an example, but the number of heating / cooling elements in the present invention is not limited to this. The plurality of heating / cooling elements la and lb are connected to a control device, and are individually controlled so that they can be set to different temperatures.

 [0036] As the heating / cooling elements la and lb, it is preferable to use Peltier elements because the heat conductor 2 can be heated or cooled according to a desired temperature. If only the heat conductor 2 is to be heated, a heater such as an electric heater can be used. Further, a Peltier element and a heater may be combined. In particular, when the temperature is set to be higher than room temperature, it is preferable to use a heater and a Peltier element together as the heating / cooling element on the high temperature side.

[0037] Further, if there is a large space between the plurality of heating and cooling elements la and lb, the central part of the heat conductor 2 may be affected by the ambient temperature, making it difficult to control the formed temperature profile. is there. In order to avoid such an effect, the force that makes the lower surface of the heat conductor 2 that is not in contact with the heating / cooling element between the heating / cooling elements la and lb be in an adiabatic state, and the distance between them can be ignored For example, it is preferable to set the length to be less than 50%, more preferably less than 10% of the length of the Peltier element measured in the direction of forming the temperature gradient. Specifically, the thickness is preferably 100 mm or less. Further, as shown in FIG. 1, the heating and cooling elements la and lb Most preferably, they are placed in close proximity to one another.

 [0038] The heat conductor 2 receives the heat (including cold heat) of the heating and cooling elements la and lb, and transfers the heat to the heat conductor 2 surface (the surface opposite to the surface in contact with the heating and cooling element; The sample 5 is placed on the sample placement surface.) (In FIG. 1, the sample 5 is held on the plate 4.) As described above, a temperature gradient is formed in the heat conductor 2 along the arrangement direction of the heating and cooling elements la and lb. In the present apparatus, by arranging a plurality of samples 5 along the direction of the temperature gradient, it is possible to adjust the temperature to different temperatures.

In the present embodiment, as described above, the heat conductor 2 has a heat conduction characteristic, specifically, specific gravity d [kgZm ³ ], specific heat cQiZ (kg′K)], and thermal conductivity When K [WZ (m'K)], the force expressed by dcZK is composed of two or more different materials.

 FIG. 2 is a cross-sectional view showing one example of the heat conductor 2 according to the present embodiment. The heat conductor 2 is made of two kinds of materials having different dcZK values. The members 2a and 2b made of each of these materials are arranged so as to be adjacent to each other along the direction of forming the temperature gradient, and the members 2a made of the material having the smaller dcZK value among the two materials are used. Are arranged at the center, and members 2b made of a material having a large dcZK value are arranged at both ends. As a result, the average value of the dcZK value can be adjusted so as to be smaller at the center of the thermal conductor in the above-mentioned region than at the end of the region where the temperature gradient is formed, thereby realizing a uniform temperature gradient. It is a character that can be.

 Further, the heat conductor 2 may be made of three or more materials having different dcZK values. FIG. 3 is a cross-sectional view showing an example in which the heat conductor is made of three kinds of materials. In this case, the members 2a, 2b, and 2c made of these three types of materials are subjected to a temperature gradient forming direction such that the dcZK value is gradually reduced from the end to the center of the heat conductor 2 as the force increases. May be arranged so as to be adjacent to each other. That is, a member 2a made of a material having the smallest dcZK value among the three materials is arranged at the center portion, and members 2b made of a material having a middle dcZK value are arranged at both ends thereof. A member 2c made of a material having the maximum dcZK value is disposed outside the member 2c.

As described above, the heat conductor 2 is configured by arranging a plurality of members in the temperature gradient forming direction. In this case, it is preferable to arrange the member having the maximum dcZK value so as to be located at a higher temperature side than the center of the heat conductor. According to this example, since the portion where the average value of the dcZK value of the heat conductor is minimum is located on the high temperature side, the temperature gradient can be further uniformed.

 In FIGS. 2 and 3, the connecting surface force between members is perpendicular to the direction in which the temperature gradient is formed, but the present invention is not limited to this. That is, the connection surface may be inclined with respect to the temperature gradient forming direction.

 FIG. 4 is a cross-sectional view showing another example of the heat conductor 2. The heat conductor 2 is made of two kinds of materials having different dcZK values, and has a structure in which layered members 2a and 2b made of these materials are laminated. In these two members, a member 2a made of a material having a small dcZK value constitutes an upper surface which is a contact surface of the heat conductor 2 with a sample, and a member 2b made of a material having a large dcZK value is formed by: The heat conductor 2 is arranged so as to constitute a lower surface which is a contact surface with the heating / cooling elements la and lb. That is, a member 2a made of a material having a small dcZK value is laminated on a member 2b made of a material having a large dcZK value. This makes it possible to realize a uniform temperature gradient while securing a desired temperature difference. In this example, the layer thickness of each member is uniform over the entire temperature gradient forming region.

FIG. 5 is a cross-sectional view showing a further preferred example in which the heat conductor 2 has a laminated structure. The heat conductor 2 is made of two kinds of materials having different dcZK values, and has a structure in which layered members 2a and 2b made of these materials are laminated. Then, of these two members, the layer thickness of the member 2a made of a material having a small dcZK value is slightly larger at the center than at the end of the heat conductor 2, and the dcZK value is large. The thickness of the layer 2 of the member 2b is adjusted to be smaller at the center than at the end of the heat conductor 2. Accordingly, the average value of the dcZK value can be adjusted so as to be smaller at the center of the thermal conductor than at the end of the region where the temperature gradient is formed, thereby realizing a uniform temperature gradient. Because it can be. Further, in the heat conductor 2, the layer thickness of both members is adjusted to complement each other, and the layer thickness of the entire heat conductor 2 is adjusted to be substantially uniform. . FIG. 6 is a cross-sectional view showing a further preferred example of the heat conductor 2 having a laminated structure. As shown in FIG. 5, the heat conductor 2 is formed by laminating two layered members 2a and 2b having different dcZK values, and has a small dc ZK value.The layer thickness of the member 2a is smaller than that of the end of the heat conductor 2. The thickness is adjusted so that it becomes larger at the center and is smaller at the center than at the end of the heat conductor 2 in the member 2b having a large dcZK value. Further, the portion (A in the figure) where the layer thickness of the member 2a having a small dcZK value is maximum and the layer thickness of the member 2b having a large dcZK value is minimum is located closer to the center of the thermal conductor 2. , It is adjusted to be located on the high temperature side. In FIG. 6, it is assumed that the heating / cooling element lb is set at a higher temperature than the heating / cooling element la. According to this example, since the portion where the average value of the dcZK value of the heat conductor 2 is minimum is located on the high temperature side, further uniform temperature gradient can be realized.

 When the heat conductor 2 has a laminated structure, the number of layers is not limited to two, but may be three or more. In this case, each layer may be composed of a different material (that is, three or more materials are used), or two materials may be alternately laminated. In any case, it is preferable that the layer thickness of each layer is adjusted so that the dcZK value (average value) distribution of the heat conductor 2 is in the form described above. It is not particularly limited.

 Further, in FIGS. 5 and 6, the force in which the layer thickness of each member continuously changes along the direction of forming the temperature gradient is not limited to this. That is, the layer thickness may change stepwise along the direction of forming the temperature gradient.

 [0049] In any of the examples, the thickness of the heat conductor 2 is not particularly limited, but is preferably a thickness that allows the heat of the heating / cooling elements la and lb to be efficiently transmitted to the sample placement surface. For example, it can be set to 11-20 mm, preferably 3-10 mm. The dimensions of the sample placement surface of the heat conductor 2 are not particularly limited, and may be appropriately set according to the purpose of use of the apparatus.

 [0050] As a material constituting the heat conductor 2, for example, a metal material such as copper, aluminum, stainless steel (SUS), or brass is preferably used. In addition, it is also possible to use heat conductive resin, glass, and the like.

[0051] Further, the members forming the heat conductor 2 are joined as long as heat conduction is ensured. The method is not particularly limited. For example, it is possible to adopt a configuration in which a thermally conductive grease is applied to the members, and the members are brought into contact with each other via the grease.

 [0052] The present device may also be configured such that another heat conductor is disposed on the heat conductor 2, and a temperature gradient is formed on the sample placement surface of the another heat conductor. The another heat conductor may have, for example, a structure in which two materials having different dcZK values are stacked. FIG. 25A shows a cross-sectional view of an example of the another heat conductor. As shown in the figure, the other heat conductor 32 is formed by alternately laminating layers 29 having a large dcZK value and layers 30 having a small dcZK value ten times. As a material of the layer 29 having a large dcZK value, for example, a polyimide tape, a resin sheet, a resin coating film, or the like can be used. As a material of the layer 30 having a small dcZK value, for example, An aluminum foil, a copper foil, a deposited film of metal or silicon, a sputtered film, or the like can be used. As shown in the plan view of FIG. 25B, when the other heat conductor 32 is disposed on the heat conductor 2, a part of the heat conductor 2 on the sample placement surface is A portion exhibiting a smaller temperature gradient can be formed. The other heat conductor is not limited to the structure shown in FIG. 25A, and may have any structure as long as a temperature gradient is formed on the sample placement surface. For example, as shown in FIG. 27, a simple structure in which two small layers 34 having a large dcZK value are laminated on a layer 34 having a small dcZK value is used.

[0053] The other heat conductor may be two or more. For example, as shown in the plan view of FIG. 28, another heat conductor 32 and a second heat conductor 36 are disposed on the heat conductor 2. As the second other heat conductor 36, for example, a thin plate having a large dcZK value such as a SUS plate or a resin plate can be used. In this way, as shown in the graph of FIG. 28B, the surface of the another heat conductor 32 has a smaller temperature gradient, and the surface of the second heat conductor 36 has a larger temperature gradient. A gradient is formed. Here, for example, in a saturation curve of a protein solution, the change in the saturation concentration with respect to the temperature on the high temperature side often increases. In the temperature region where the saturation curve rises as described above, if data can be obtained densely, the value of the data is further increased. Therefore, if a protein crystallization apparatus described later including a temperature control apparatus using a configuration as shown in FIG. 28A is used, by reducing the temperature gradient on the high temperature side, the region where the saturation curve rises can be improved. Get data secretly You can do it.

 Further, in the present apparatus, a temperature measuring element 6 is provided in the heat conductor 2. As the temperature measuring element 6, for example, a temperature sensor such as a thermocouple can be used. The temperature measuring element 6 can be provided, for example, so as to correspond to each of the heating and cooling elements la and lb. This temperature measuring element 6 is connected to a control device. The controller compares the measured value input from the temperature sensor 6 with the set temperature, calculates a control amount according to the difference, and applies the control amount to the heating / cooling elements la and lb based on the control amount. Control the current flowing. As a result, the heating / cooling elements la and lb can be controlled so as to quickly reach the set temperature or can be controlled so as to correct the deviation from the set temperature.

 [0055] The temperature measuring element 6 is preferably embedded in the sample disposition surface of the heat conductor 2. By providing the temperature measuring element 6 on the sample placement surface, the difference between the set temperature and the actual sample temperature can be reduced.

 FIG. 18 shows an example of a method of embedding the temperature measuring element 6 in the heat conductor 2. FIG. 18 is a cross-sectional view of the heat conductor 2 on a sample placement surface. When the temperature measuring element 6 is, for example, a thermocouple with a protective tube, a platinum resistance temperature measuring element, or the like, a temperature sensor is provided at a tip of the protective tube in the temperature sensor. Is common. Therefore, when the above-described temperature sensor (thermometer) is embedded in a heat conductor and used, as shown in FIG. A thermometer insertion hole 19 with a depth up to approximately the center of the heat conductor 2 is formed in the sample placement surface of the heat conductor 2 so as to be approximately at the center, and the thermometer 6 can be inserted into this hole. preferable.

 FIG. 19 shows another example of a method of embedding the temperature measuring element 6 in the heat conductor 2. FIG. 19 is a cross-sectional view of the heat conductor 2 on a sample placement surface. In the method shown in FIG. 18, since the heat conductor 2 has poor symmetry in the depth direction, the heat flow is disturbed, and the temperature uniformity in the depth direction of the heat conductor 2 (the direction perpendicular to the direction in which the temperature gradient is formed) is obtained. May adversely affect sex. In such a case, as shown in FIG. 19, a temperature measuring element 6 having a temperature sensing portion 20 disposed substantially at the center thereof is prepared, and a temperature measuring element insertion hole 19 is provided on the sample placement surface of the heat conductor 2. It is more preferable that the temperature measuring element 6 is formed so as to penetrate the heat conductor 2 and the temperature measuring element 6 is inserted therein.

Further, in the present apparatus, another temperature measuring body for temperature monitoring is provided in the heat conductor 2. May be. FIG. 22 shows an example of installation of the another temperature measuring element. FIG. 22 is a cross-sectional view of the apparatus in a direction perpendicular to the direction in which the temperature gradient is formed. In FIG. 22, the same members as those in FIG. As shown in the figure, another temperature measuring element 37 is buried on the sample placement surface of the heat conductor 2, and the other temperature measuring element 37 is held by the temperature measuring element holder 28. At this time, it is preferable that the other temperature measuring element 37 is embedded in the end of the heat conductor 2 so as not to affect the thermal characteristics of the heat conductor 2 as much as possible. As the another temperature measuring element 37, for example, the same one as the temperature measuring element can be used. Further, from the viewpoint of preventing heat inflow and outflow from the environment into the other temperature measuring element 37, it is preferable that the temperature measuring element holder 28 be made of a material having low heat conductivity and high material strength. ,.

 FIG. 21 shows a cross-sectional view of another example of the temperature control device according to the first embodiment of the present invention. The temperature control device shown in FIG. 21 has a lid 26 having a multilayer structure with an air layer interposed between the temperature control device shown in FIG. The lid 26 has a multilayer structure in which two transparent acryl plates 24 sandwich an air layer 25. The two transparent acrylic plates 24 are joined using, for example, an O-ring 27. When the lid is arranged on the upper surface of the temperature controller shown in FIG. 1, when the heat conductor 2 is kept at a low temperature, in a high humidity environment, dew condensation may occur on the lid. In such a case, as shown in FIG. 21, it is preferable to prevent the occurrence of dew condensation by forming the lid with a lid 26 having a multilayer structure with an air layer interposed therebetween. The lid 26 has a multi-layer structure sandwiching the air layer, so that the temperature difference between the upper surface of the lid and the lower surface of the lid becomes large, and the occurrence of dew condensation can be prevented. The material of the lid 26 having the multilayer structure is not limited to the above-mentioned transparent acrylic plate, but may be any other material. In this case, it is preferable to use a transparent material in consideration of the visibility inside the device. Further, the layer sandwiched between the materials is not limited to the above-described air layer, and may be a layer made of another material having high heat insulating properties. Also in this case, it is preferable to use a material having high transparency in consideration of the visibility inside the device.

 Next, the operation of the above device will be described.

[0061] First, the controller is operated to set the temperatures of the heating and cooling elements la and lb to different temperatures. This set temperature can be appropriately set according to the purpose of use of the apparatus, which is not particularly limited. As an example, the dissolution of protein crystals When used for adjusting the liquid temperature, the temperatures of the heating and cooling elements la and lb are set, for example, within the range of 5 to 40 ° C, preferably 4-1 to 20 ° C, and the temperature difference between the two is set to For example, the temperature can be 2 to 40 ° C, preferably 5 to 20 ° C.

 [0062] The heat of the heating / cooling element is conducted to the heat conductor and further conducted in the heat conductor 2. At this time, a temperature gradient is formed in the heat conductor as described above. In this state, when the sample is placed on the heat conductor sample placement surface, the temperature of the sample is adjusted according to the temperature of the heat conductor 2 at the place where the sample is placed. At this time, since a temperature gradient is formed on the surface of the heat conductor, the sample temperature to be adjusted differs depending on the arrangement position of the sample. Therefore, when a plurality of samples are arranged and placed along the direction of forming the temperature gradient, experiments and the like under different temperature conditions can be performed simultaneously by one operation.

 Further, according to the present embodiment, the temperature linearly increases as the distance from one end of the heat conductor along the direction of forming the temperature gradient increases. A temperature profile that rises or falls can be achieved. Therefore, the predetermined position of the heat conductor can be easily adjusted to the target temperature.

 [0064] In the present apparatus, the number of the temperature gradient generating units may be one or two or more.

 When the number of the temperature gradient generation units is two or more, it is preferable that a plurality of the temperature gradient generation units are connected to one control device as shown in FIG. In this way, space can be saved and the experimental efficiency can be improved.

 [0065] Further, the temperature gradient generating unit can prevent dew condensation from occurring when the temperature is kept low, for example, by covering with a heat insulating material. However, in a high humidity environment, dew condensation is completely generated. It can be difficult to prevent. In such a case, as shown in FIG. 20, it is preferable to dispose the temperature gradient generating unit 21 above a power control device such as a power supply 22 and a control board 23. In this way, since the power supply 22 or the control board 23 emits heat during operation or the like, the surface of the heat insulating material that covers the temperature gradient generation unit 21 disposed above the power supply 22 or the control board 23 generates the heat. As a result, warm and dry air flows, and as a result, it is possible to prevent the occurrence of dew condensation. In FIG. 20, arrows indicate the flow of the warm and dry air.

(Second Embodiment) In the present embodiment, the heat conductor is made of one or more materials, and a plurality of recesses are formed on the surface of the heat conductor opposite to the sample placement surface, and air is allowed to exist in the recesses. The configuration will be described. The temperature controller according to the present embodiment has the same configuration as the first embodiment except that the configuration of the heat conductor is different.

 FIG. 7 is a cross-sectional view showing one example of the heat conductor according to the second embodiment of the present invention. The heat conductor 2 is made of one kind of material, and has a plurality of recesses 8 formed on the back surface (the surface on the heating / cooling element side). It should be noted that air exists inside the recess 8.

 [0068] The shape of the concave portion 8 is not particularly limited, and may be, for example, a groove shape, a hole shape, or the like. However, the dimension of the recess 8 in the direction of forming the temperature gradient is preferably set to 0.5 to 20 mm, and more preferably to 15 to 15 mm. This is because excessive uniformity of the temperature in the heat conductor 2 due to convection of air generated in the direction of forming the temperature gradient can be suppressed. For this reason, when the recess 8 is formed as a groove, it is preferable to form the groove 8 as a groove extending in a direction perpendicular to the temperature gradient forming direction.

 In this heat conductor, as shown in FIG. 7, in the region where the temperature gradient of the heat conductor 2 is formed, the depth of the recess 8 is such that the end force of the region and the end force of the region are closer to the center. It has been adjusted to be smaller. By changing the depth distribution of the recess 8 along the temperature gradient forming direction in this way, the average value of the dcZK value is adjusted to be smaller at the center of the region than at the end of the region. This is because the temperature gradient can be made uniform. Note that, in the case of this embodiment, the plurality of recesses 8 can be regularly arranged so that the number of the recesses 8 per unit area is uniform in the region as shown in FIG. It is.

 [0070] Furthermore, in the heat conductor having such a configuration, in the temperature gradient forming region of the heat conductor 2, the portion where the depth of the concave portion 8 is minimum is located at the center of the region. It is preferable to adjust so as to be located on the higher temperature side. This is because the portion where the average value of the dcZK value of the heat conductor 2 is minimum is located on the high temperature side, so that a further uniform temperature gradient can be realized.

Further, as another example of the heat conductor according to the present embodiment, in a region where a temperature gradient is formed, the number of recesses per unit area increases from the end to the center of the region. Well, It is also possible to adopt a form formed to reduce the number. As this form, for example, when the concave portion has a groove shape, there can be mentioned a form in which the interval between the grooves is adjusted so as to increase from the end to the center of the region. By changing the distribution density of the concave portions along the direction of forming the temperature gradient in this manner, it is possible to adjust the average value of the dcZK value to be smaller at the center of the region than at the end of the region. This is because the temperature gradient can be made uniform. In this case, the depths of the plurality of concave portions can be adjusted so as to be equal.

 Further, in the heat conductor having such a configuration, in the temperature gradient forming region of the heat conductor, a portion where the number of the concave portions per unit area is minimized is a portion of the region. It is preferable to be located on the higher temperature side than in the center. This is because the location where the average value of the dcZK value of the thermal conductor is minimum is located on the high temperature side, so that a further uniform temperature gradient can be realized.

 [0073] It is also possible to combine the above two forms. That is, both the depth distribution and the density distribution of the concave portion may be changed along the temperature gradient forming direction of the heat conductor.

 [0074] Even in the case of the deviation, the dimensions of the entire heat conductor are not particularly limited, and can be set in the same manner as in the first embodiment.

 [0075] Also, the material constituting the heat conductor is not particularly limited, and the same material as that of the first embodiment can be used. Further, in the above description, the heat conductor is made of one kind of material, and a form in which a concave portion is provided is exemplified. However, the present embodiment is not limited to this. As in the first embodiment, two or more kinds of materials are used as the heat conductor and the concave portion is provided, so that the temperature profile can be adjusted with higher accuracy.

 [0076] Even when the thermal conductor having such a form is used, the same effect as that of the first embodiment can be achieved. Note that the operation of the above device is the same as that of the first embodiment, and a description thereof will be omitted.

 (Third Embodiment)

In the temperature control device of the present invention, a groove (groove 9 in FIG. 8) extending in a direction perpendicular to the direction in which the temperature gradient is formed is formed on the surface of the heat conductor that is in thermal contact with the sample. Preferably, it is formed. FIG. 8 is a cross-sectional view showing an example of a preferred embodiment of such a heat conductor.

 According to such an embodiment, on the sample placement surface of the heat conductor 2, the movement of heat in the direction perpendicular to the grooves 9 (that is, the direction in which the temperature gradient is formed) is suppressed. The temperature difference between this region and the region between the adjacent grooves 9 can be increased. On the other hand, in the region between the same grooves 9, heat transfer in the direction along the grooves 9 (that is, the direction perpendicular to the direction in which the temperature gradient is formed) is hindered, so that the isothermal property is improved. be able to. As described above, a large temperature difference is formed on the surface of the heat conductor 2 in the temperature gradient forming direction, and the temperature at each position on the surface of the heat conductor 2 is maintained substantially uniform in the direction perpendicular to the direction. Becomes possible. Therefore, by arranging a plurality of samples on the surface of the heat conductor 2 in the direction in which the temperature gradient is formed and also in the direction perpendicular thereto, while performing a plurality of experiments under different temperature conditions, and Multiple experiments at the same temperature can proceed at the same time.

 The shape of the groove 9 is not particularly limited. In addition to a groove having a rectangular cross section as shown in FIG. 8, a V-shaped groove, a U-shaped groove, or the like can be used. is there.

 [0080] The number of grooves 9 is not particularly limited, and can be appropriately selected depending on the purpose of use. For example, when a plate 4 including a plurality of wells is placed as shown in FIG. 8, the number of grooves 9 may be set corresponding to the number of well rows. The number of grooves 9 is, for example, 3 to 50, preferably 5 to 25.

 [0081] The width of the groove 9 and the interval between the grooves 9 can be appropriately selected according to the purpose of use, which is not particularly limited. For example, when a plate 4 including a plurality of wells is placed as shown in FIG. 8, the width of the groove 9 is set to correspond to the space between the wells, and the space between the grooves 8 is set to the size of each well. What is necessary is just to set it correspondingly. The width of the groove 9 is, for example, 0.5 to 20 mm, preferably 1 to 10 mm, and the interval between the grooves 9 is, for example, 1 to 50 mm, preferably 1 to 20 mm.

 The depth of the groove 9 is not particularly limited, but is, for example, 0.5 to 10 mm, and preferably 115 to 5 mm.

[0083] The heat conductor according to the present embodiment has a configuration in which a groove is formed in the sample arrangement. Outside, the configuration can be the same as that of the first and second embodiments. Needless to say, the same effects as those of the first and second embodiments can be achieved by the heat conductor according to the present embodiment.

(Fourth Embodiment)

 In the present embodiment, a mode in which the temperature profile can be changed with time will be described.

 FIG. 23 is a schematic diagram showing an example of a part of a temperature control device according to the fourth embodiment of the present invention. In this device, a control device such as a personal computer (PC) or a sequencer has a preset temperature setting schedule, and outputs temperature setting information to a temperature control circuit via a communication interface according to the schedule. The temperature output circuit outputs, to a control output generation circuit, a temperature control signal calculated by calculating a difference between a set temperature and a temperature at a control point obtained by a temperature sensor such as a temperature sensor. The control output generation circuit outputs a control output according to the control signal to a heating / cooling element such as a Peltier element.

 [0086] The output of the temperature control circuit is preferably a temperature control signal calculated based on a PID control parameter. The control output of the control output generation circuit output according to the temperature control signal may be a continuous current output or a constant voltage variable pulse output.

 In the device shown in FIG. 23, the voltage or current of a temperature sensor such as a temperature sensor is sent to a sensor output measuring circuit, and the sensor output measuring circuit reads the temperature indicated by the sensor, Feedback is provided to circuits, communication interfaces, and control devices such as PCs and sequencers.

 In FIG. 23, the number of the temperature control circuit, the control output generation circuit, the heating / cooling element such as the Peltier element, the temperature sensor such as the temperature sensor, and the number of sensor output circuits are each two. However, the number is not limited to three or more.

According to the present embodiment, temperature control as shown in FIG. 24 becomes possible. FIG. 24A is a graph showing an example in which a constant temperature force also gradually increases the temperature gradient, FIG. 24B is a graph showing an example in which the temperature is lowered as a whole while maintaining a constant temperature gradient, and FIG. average It is a graph which shows the example which changes a temperature gradient gradually, without changing a temperature. Further, by combining the temperature control described in the above example, various temperature controls corresponding to the test object can be performed, for example, a temperature drop after a temperature rise.

 The device according to the present embodiment can have the same configuration as the first, second, and third embodiments except that a part of the device includes the configuration shown in FIG. In addition, it goes without saying that the same effects as those of the first, second, and third embodiments can also be achieved by the device according to the present embodiment.

 [0091] (Fifth Embodiment)

 The temperature control device of the present invention can be used for all experimental operations that require setting of temperature conditions, such as treatment involving various chemical reactions and measurement such as evaluation of temperature dependence of physical properties.

 [0092] The temperature control device of the present invention can be suitably used for temperature control of a sample solution containing a protein, for example, in crystallization of a protein. In addition, protein crystallization includes a batch method, a dialysis method, a vapor diffusion method, and the like, and the temperature control device of the present invention can be applied to this deviation method. .

 FIG. 9 is a cross-sectional view showing one example of a protein crystallization device to which the temperature control device of the present invention is applied. This apparatus is a protein crystallization apparatus suitable for the diffusion method, particularly the hanging drop method. In FIG. 9, the same members as those in FIG. 1 are denoted by the same reference numerals, and description of those members will be omitted.

 [0094] The present apparatus includes the above-described temperature control apparatus of the present invention, a holding mechanism 11 for holding a sample, and a housing 10 surrounding these. The temperature controller is arranged with the sample arrangement surface of the heat conductor 2 facing downward, and the holding mechanism 11 is arranged below the arrangement surface. Further, the holding mechanism 11 has a sample holding surface for holding a sample, and an elevating mechanism for raising and lowering this surface. The sample holding surface is disposed so as to be opposed to the sample placement surface of the heat conductor 2 of the temperature controller at an appropriate distance.

 [0095] Next, the operation of this device will be described.

In this apparatus, the sample solution is set on the holding mechanism 11 while being held on the plate 4. This plate 4 has a plurality of well plates and a well opening. It is composed of a cover plate that is arranged on the Pell plate so as to close it. The reservoir holds the reservoir solution containing the precipitant. The sample solution containing the protein is placed as droplets on the inner surface of the cover plate. By arranging the cover plate on the well plate, a droplet of the sample solution is dripped on the upper part of the well containing the reservoir solution. That is, the sample solution and the reservoir solution are stored in the closed space. In the plate 4, the volatile components contained in the sample solution and the reservoir solution are diffused between the solutions, and as a result, the crystallization of the protein proceeds in the sample solution.

[0097] In the plate 4, the plurality of wells are preferably arranged two-dimensionally. In this case, it is possible to perform crystallization under different temperature conditions in one operation by arranging the plate in the apparatus so that one of the arrangement directions is along the direction of forming the temperature gradient of the temperature control apparatus. Become. Furthermore, if the plate 4 is arranged in the apparatus so that the other arrangement direction is along the direction perpendicular to the direction of forming the temperature gradient of the temperature control apparatus, a plurality of experiments can be performed under the same temperature conditions. . The number of pells is not particularly limited.

 [0098] The plate 4 is arranged on the sample holding surface of the holding mechanism 11, and the holding mechanism 11 is operated to raise the sample holding surface toward the heat conductor 2. Thereby, the upper surface of the plate 4, that is, the cover plate comes into contact with the heat conductor 2 of the temperature controller. The temperature control device operates as described in the first embodiment, and the temperature of the sample solution droplets hanging on the cover plate is controlled according to the temperature of the heat conductor 2. At this time, the temperature of the sample solution on each well arranged along the temperature gradient forming direction of the heat conductor 2 is adjusted to a different temperature according to the temperature gradient. Therefore, crystallization under different temperature conditions can be performed by one operation.

 [0099] The set temperature in the temperature control device is not particularly limited, but the heating / cooling element la set to the highest temperature is, for example, 10 to 100 ° C, preferably 10 to 50 ° C. For example, the heating / cooling element lb set to a low temperature may be set to 5 to 30 ° C, preferably 412 to 20 ° C.

[0100] After the crystallization is completed, in the above apparatus, the holding mechanism 11 is operated to By lowering the sample holding surface, the plate 4 and the heat conductor 2 are separated. Then, the plate 4 is also taken out of the apparatus, and the crystals are taken out of the plate 4, thereby completing the protein crystallization operation.

 [0101] In this device, the plate for holding the sample solution is arranged with respect to the temperature control device by the elevating operation of the holding mechanism. That is, by making the holding mechanism movable, it is possible to realize the arrangement of the plates with the temperature control device fixed. In this way, it is not necessary to make the temperature control device, which is the core of the protein crystallization device, movable, so that it is possible to avoid external shocks and the like that this device receives as it moves, and the possibility of device failure. Can be reduced.

 (Sixth Embodiment)

 Next, another example of the protein crystallization apparatus to which the temperature control apparatus of the present invention is applied will be described. This device is particularly suitable for the sitting drop method.

 [0103] The present device includes the temperature control device of the present invention as described above. When the heating / cooling element and the heat conductor constituting the temperature control device are a first heating / cooling element and a first heat conductor, the second heating / cooling element and the second heating / cooling element A second heat conductor that is in thermal contact with the cooling element and is thermally insulated from the first heating / cooling element and the first heat conductor; The cooling element keeps the second heat conductor at a constant temperature.

 FIG. 10A is a cross-sectional view showing an example of such a protein crystallization apparatus. Further, FIG. 1OB is a cross-sectional view of the above-mentioned protein crystallization apparatus, in which the cross-sectional direction is orthogonal to the cross-sectional view of FIG. 10A. This device includes a plurality of first heating / cooling elements la and lb, and a first heat conductor 2 arranged in thermal contact with the upper surface thereof. Note that these components are as described in the embodiment of the temperature control device of the present invention, and description thereof will be omitted.

[0105] Below the first heating and cooling elements la and lb, a second heating and cooling element 14 is arranged. The second heating / cooling element 14 has a heat acting surface (a surface for heating / cooling the object) larger than the first heating / cooling element la, lb. 1 heating and cooling elements la and lb are arranged. The heat generated by the second heating / cooling element 14 The second heat conductor 12 has a configuration in which the second heat conductor 12 is in thermal contact with an area of the application surface that is not covered by the first heating / cooling elements la and lb. On the heat working surface of the second heating / cooling element 14, the area not covered by the first heating / cooling element la, lb is, for example, as shown in FIG. , lb. In such an embodiment, the second heat conductor 12 is arranged at both ends of the first heat conductor 2 so as to sandwich the first heat conductor 2 therebetween.

 [0106] As the second heating / cooling element 14, it is preferable to use a Peltier element as in the case of the first heating / cooling elements la and lb. If only the heat conductor 12 is heated, a heater such as an electric heater can be used.

 [0107] The second thermal conductor 12 is not particularly limited, but it is preferable to use a material having a small dcZK value. For example, metal materials such as copper, aluminum, stainless steel (SUS), and brass can be used. The dimensions are not particularly limited, and can be appropriately set according to the dimensions of the plate used.

 A heat insulator 13 is arranged between the second heat conductor 12, the first heating / cooling elements la and lb, and the first heat conductor 2. The heat insulator 13 is not particularly limited. For example, fluorine resin, heat insulating ceramics, styrofoam, silicon rubber, or the like can be used.

 Further, the present apparatus is provided with a control device (not shown) for driving and controlling the heating / cooling elements la, lb and 14. This controller individually controls each of the heating and cooling elements. Further, on the lower surface of the second heating / cooling element 14, a heat bath section 3 for recovering exhaust heat is arranged.

 [0110] Next, the operation of this device will be described.

[0111] In the present apparatus, the sample solution is placed in the apparatus while being held on the plate 4. The plate 4 includes a first plate 4a having a first space 4a and a second plate 4b having an internal space communicating with each other, and a cover plate disposed on the second plate so as to close the opening of the second plate 4a. It is composed of The sample solution is held in the first well 4a. The second well 4b holds a reservoir solution containing a precipitant. By arranging the cover plate on the well plate, the cover plate can be closed as in the fourth embodiment. The sample solution and the reservoir solution are housed in the closed space.

 [0112] In the plate 4, one cell is constituted by one first well 4a and one second well 4b, and the cells are arranged two-dimensionally. Specifically, as shown in FIG. 10, a configuration in which cells are arranged in two rows can be employed. At this time, the cell rows are arranged such that the first pegs 4a are adjacent to each other. With this arrangement, it is possible to arrange the first jewel 4a at the center of the plate 4 and the second jewel 4b at both ends. It should be noted that the number of cells constituting the cell column is not particularly limited.

 [0113] The plate 4 is arranged on the surface of the heat conductor constituting the device. At this time, as shown in FIG. 10, the first well 4a is in thermal contact with the first thermal conductor 2, and the second well 4b is in thermal contact with the second thermal conductor 12. Placed in

 [0114] As described in the embodiment of the temperature control device of the present invention, by setting the first heating and cooling elements la and lb to different temperatures on the surface of the first heat conductor 2, A temperature gradient is formed. Then, the temperature of the sample solution held in the first well 4 a is adjusted according to the temperature of the first heat conductor 2. At this time, the sample solutions in the first wells 4a arranged along the temperature gradient forming direction are adjusted to different temperatures according to the temperature gradient. Therefore, crystallization under different temperature conditions can be performed by one operation.

 [0115] The set temperatures of the first heating and cooling elements la and lb can be set in the same manner as in the fifth embodiment, which is not particularly limited.

 On the other hand, second heat conductor 12 receives heat (including cold heat) of second heating / cooling element 14 and is maintained at a constant temperature. Then, the temperature of the reservoir solution held in the second well 4b is adjusted according to the temperature of the second heat conductor 12. At this time, a temperature gradient is not substantially formed on the surface of the second heat conductor 12. Therefore, with respect to the plurality of second wells 4b formed on the plate 4, it is possible to maintain all the reservoir solutions held therein at an isothermal temperature.

[0117] The set temperature of the second heating / cooling element 14 is not particularly limited, but may be, for example, 5 to 50 ° C, preferably 412 to 20 ° C. [0118] After the crystallization is completed, the plate 4 is also taken out of the apparatus, and the crystal is taken out of the plate 4, thereby completing the protein crystallization operation.

[0119] As described above, the above-described device has an advantage that the sample solution can be adjusted to different temperatures while maintaining the reservoir solution of each cell at an isothermal temperature with a relatively simple device configuration. I have.

 Example 1

 [0120] A temperature controller having a configuration as shown in Fig. 1 was produced. As shown in FIGS. 11A and 11B, two types of heat conductors were manufactured using aluminum-palladium and SUS (Examples 11 and 12). Each of the heat conductors has a structure in which a triangular prism-shaped aluminum member 15 is arranged at the upper part and two triangular prism-shaped or square prism-shaped SUS members 16 are arranged at the lower part. The contact was made through a neutral grease. The overall dimensions of the thermal conductor were 110 mm wide, 60 mm deep and 6 mm thick. That is, the thickness of the SUS member 16 at the center of the heat conductor is Omm, and the thickness of the SUS member 16 at both ends is 6 mm. Then, in Example 1-1, the width (W1) of the aluminum member 15 was set to 110 mm, and in Example 1-2, the width (W2) of the aluminum member 15 was set to 66 mm. In each case, the effective area for forming the temperature gradient was set to 60 x 30 mm.

 As a comparative example, as shown in FIG. 11C, a heat conductor composed only of the SUS member 16 was produced. Also in this comparative example, the overall dimensions of the heat conductor were 110 mm in width, 60 mm in depth, and 6 mm in thickness, and the effective area for forming the temperature gradient was 60 × 30 mm.

 Next, two Peltier elements (size: 40 mm × 40 mm) are prepared, arranged adjacent to each other, and connected to a control device configured so that both elements can be controlled individually. Continued. Then, the thermal conductor was disposed above both Peltier elements. Thermocouples were provided on portions of the heat conductor surface corresponding to both Peltier devices. In order to reduce the influence of heat radiation to the surrounding environment, a heat insulating cover made of fluorine resin was provided on the side of the Peltier element and the heat conductor. Further, a water-cooling unit was arranged as a heat bath section on the lower surface of the Peltier device to obtain a temperature controller.

[0123] In the above temperature control device, the set temperatures of the two Peltier elements are set to 5 ° C and 15 ° C, 2.5 ° C and 17.5 ° C, 0 ° C and 20 ° C, respectively. Depending on the type of setting, The water temperature in the well of the placed plate was measured, and the temperature gradient obtained at each position of the measured force was calculated. The difference between the water temperature in the high-temperature end and the water temperature in the low-temperature end is defined as the generation temperature difference, and the value obtained by dividing the water temperature difference between adjacent wells by this generation temperature difference is the temperature gradient at the intermediate position between the adjacent wells. Relative values were used. The measurement was performed with the above three settings, and the average value of the temperature gradient obtained for each location was obtained.

[0124] The results are shown in FIG. In FIG. 12, the position on the upper surface of the heat conductor is represented by a reference number (Omm) at the center of the heat conductor, a lower temperature side of the reference position as "1", and a higher temperature side as "+", and a temperature. It shall be represented by a numerical value indicating the distance from the center along the gradient forming direction.

 [0125] As shown in Fig. 12, when the heat conductor of the comparative example was used, the central force where the temperature gradient was large at the center of the heat conductor (near position 0 mm) became smaller as the distance from the conductor increased. It was confirmed that the temperature gradient was non-uniform. In contrast, when the heat conductors of Example 11 and Example 1-2 were used, it was confirmed that the uniformity of the temperature gradient was improved as compared with the comparative example. In particular, when the heat conductor of Example 12 was used, it was confirmed that a substantially uniform temperature gradient was obtained over the entire temperature gradient forming region.

 Next, the plate holding the sample was placed in the well on the temperature controller using the heat conductor of Example 1-1, and the set temperatures of the two Peltier devices were each set to 5 °. The temperature was set at C and 15 ° C. Water was used as a sample, and the amount was 10 / zL for each well. The plate used is 9 x 5 with a 3 mm diameter jewel force and a pitch of 7 mm. The plate has 9 directional forces. The directional force is parallel to the direction in which the Peltier elements are arranged (temperature gradient forming direction). As above, placed on a heat conductor.

 [0127] After 12 hours, the water temperature in each well was measured. The results are shown in FIG. As for the sample numbers, the numbers are assigned to the nine low-temperature-side forces applied to the nine wells arranged along the temperature gradient forming direction. In the above plate, the nine wells are arranged at regular intervals along the temperature gradient forming direction. Therefore, the horizontal axis in FIG. 13 can be regarded as the distance along the temperature gradient forming direction.

As shown in FIG. 13, according to the temperature control device, the temperature of the water held in the plurality of wells could be simultaneously set to different temperatures. Furthermore, this temperature control It was confirmed that the temperature of the device can be changed almost linearly with respect to the distance along the direction of forming the temperature gradient.

 Next, the relationship between the control set temperature of the control device and the temperature generated on the heat conductor in the temperature control device using the heat conductor of Example 1-2 was examined.

[0130] FIG. 17A shows the average value of the high temperature set temperature and the low temperature set temperature of the control device (the set average temperature (° C

3) is a graph showing the relationship between the measured average temperature (° C.) and the central part of the heat conductor. As shown, both are in a proportional relationship.

FIG. 17B is a graph showing the relationship between the difference between the high temperature setting temperature and the low temperature setting temperature (the setting temperature difference (K)) of the control device and the generated temperature gradient (KZmm). As shown, they are in a proportional relationship.

 [0132] From the three parameters of the gradient and the y-intercept of the graph shown in Fig. 17A and the gradient of the graph shown in Fig. 17B, the central temperature (° C) and the center of the temperature gradient forming region of the heat conductor are obtained. And the generated temperature gradient (KZmm) was determined. Further, the temperature at a predetermined position in the temperature gradient forming region of the heat conductor was estimated using the following equation (1).

 [0133] Y (° C) = AX + B (1)

 Y: temperature at a predetermined location in the temperature gradient forming area of the heat conductor

 X: Generated temperature gradient (KZmm)

 A: Distance from the center of the temperature gradient forming area to the predetermined location (mm)

 T: Central temperature of the temperature gradient forming area (° C)

In the heat conductor used in Example 12 (heat conductor shown in FIG. 11B), there was a portion where the joint between aluminum member 15 and SUS member 16 was exposed on the upper surface of the heat conductor. From there, the thermal conductive grease may seep out and contaminate the sample placed above the thermal conductor.

[0135] In such a case, if a heat conductive adhesive is used instead of the heat conductive grease, contamination of the sample can be prevented. However, when using the heat conductive adhesive, care must be taken since the bonding operation must be completed quickly and forcely within the solidification time after the application of the adhesive. It is also necessary to pay attention to the uniformity and thickness of the adhesive layer, the relative position between the aluminum member 15 and the SUS member 16, and the like. [0136] Even in the case of using the heat conductive grease, as shown in Fig. 16, the sample placement surface of the heat conductor is covered with another heat conductor 17, so that the sample placement is achieved. Sample contamination without changing the temperature profile of the surface can be prevented. In FIG. 16, reference numeral 18 denotes a temperature measuring element, and the same members as those in FIG. 11B are denoted by the same reference numerals. As the further heat conductor 17, for example, it is preferable to use a thin plate having a large dcZK value (for example, a SUS plate having a thickness of 0.5 mm). As a result, in the case of using a thermally conductive durse, even if the application of the thermal conductor is not limited to a certain extent and the application of the thermal grease is not uniform, the grease will spread during the running-in operation. Can be easily assembled.

 Example 2

 The protein was crystallized by the following procedure using the temperature controller (the device using Example 1-1 as a heat conductor) prepared in Example 1 above. Crystallization was carried out by a stationary batch method.

 [0138] Chicken egg white lysozyme (Hen Egg White Lysozyme) was dissolved in a 0.1 M acetate buffer solution of pH 4.5 to a concentration of 25 mgZmL. Further, 25 mg ZmL of sodium salt was mixed to prepare a sample solution. 10 L of this solution was placed in each well of the plate. The same plate as in Example 1 was used.

 [0139] The above plate was placed on the heat conductor (Example 1-1) of the temperature controller manufactured in Example 1. The set temperatures of the two Peltier devices were set to 5 ° C and 15 ° C, respectively. The mounting of the plate was performed so as to be parallel to the direction in which the directional force Peltier elements (in which the nine gradients were arranged) (temperature gradient forming direction) were arranged.

 [0140] After one week, the device power plate was taken out, and the crystals formed in each well were observed with an optical microscope. The results are shown in FIG. The sample numbers are given to the nine wells arranged along the temperature gradient forming direction in the order of the applied low-temperature side force.

[0141] Further, when the liquid temperature in each of the gels was measured, the temperature of the solution held therein could be simultaneously set to different temperatures for a plurality of wells, as shown in FIG. In addition, the temperature changes almost linearly with respect to the distance along the temperature gradient forming direction. Was confirmed.

 Example 3

 [0142] On a lower surface of a SUS flat plate having a width of 110mm, a depth of 60mm, and a thickness of 6mm, a plurality of grooves having a pitch of 3mm and a width of 1mm were formed, and a heat conductor having a configuration as shown in Fig. 7 was produced. The effective area for forming the temperature gradient was 60 X 30 mm, which corresponded to 9 X 5 plates with 3 mm diameter holes arranged at 7 mm pitch. Then, a temperature controller was manufactured in the same manner as in Example 1 except that this heat conductor was used and a Peltier element and a ceramic heater were used in combination as a heating / cooling element on the high temperature setting side.

 [0143] In the above temperature control device, with the heaters not operating, the set temperatures of the two Peltier elements were set to 5 ° C and 15 ° C, respectively, and set on the upper surface of the heat conductor. The temperature at each position of the well of the prepared plate was measured. The results are shown in FIG. 15A. Also, set the heater to 50 ° C, set the set temperatures of the two Peltier elements to 5 ° C and 50 ° C, respectively, and set the temperature at each position of the plate on the top surface of the heat conductor. Was measured. The results are shown in FIG. 15B. In FIG. 15, the position on the upper surface of the heat conductor is indicated by a reference number (Omm) at the center of the heat conductor, a lower temperature side of the reference position as `` 1 '', and a higher temperature side as `` + '', and a temperature. It shall be represented by a numerical value indicating the distance from the center along the gradient forming direction.

 [0144] As shown in FIGS. 15A and 15B, it was confirmed that a temperature gradient could be formed on the upper surface of the heat conductor. In addition, it was confirmed that the temperature of the upper surface of the heat conductor can be changed almost linearly with the distance along the direction of forming the temperature gradient.

 Example 4

 [0145] Another heat conductor 32 having the structure shown in Fig. 25A was prepared. Polyimide tape was used for the layer 29 having a large dcZK value, and aluminum foil was used for the layer 30 having a small dcZK value. Next, as shown in the plan view of FIG. 25B, the another heat conductor 32 was disposed on the heat conductor 2 having a thickness of 1.5 mm. The heat conductor 2 used was the same as the heat conductor used in Example 12 above. Except for this, the temperature controller was manufactured in the same manner as in Example 1.

[0146] The graph of FIG. 26 shows the results of measurement of the in-wall temperature of the surface of another heat conductor 32 and the in-wall temperature of the surface of the heat conductor 2. As shown, the temperature of a portion of another heat conductor 32 is The temperature gradient was approximately 1Z2 of the temperature gradient in the part without another heat conductor 32. Next, a temperature controller was manufactured using another heat conductor having the structure shown in FIG. 27 instead of the another heat conductor 32. Silicon rubber was used for the layer 34 having a large dcZK value, and an aluminum thin plate was used for the layer 33 having a small dcZK value. Even in this case, the temperature gradient in a part without another heat conductor could be reduced to about 60% of the temperature gradient in a part without another heat conductor.

 Example 5

 [0147] As shown in Fig. 28A, one having three heat conductor forces was also prepared. In this configuration, another heat conductor 32 was the same as the other heat conductor used in Example 4, and a second other heat conductor 36 was a SUS plate. Except for this, the temperature controller was manufactured in the same manner as in Example 4. In the above configuration, the same heat conductor 2 as that used in Example 4 was used as the heat conductor 2. In the above temperature controller, as shown in FIG. 28B, a smaller temperature gradient is applied to the surface of the another heat conductor 32, and a larger temperature gradient is applied to the surface of the second heat conductor 36. Been formed.

 Industrial applicability

[0148] The temperature controller of the present invention can be used for any operation that requires setting of temperature conditions, such as an operation involving a chemical reaction such as substance synthesis, measurement of a biochemical reaction, and measurement of temperature dependence evaluation of physical properties. It can be used for operation and can be applied to a wide range of applications in various industrial fields. In particular, it is useful for applications that require treatment under various temperature conditions, such as chemical reactions used in combinatorial chemistry, enzymatic reactions, crystallization of proteins, and temperature screening in microbial culture. The protein crystallization apparatus of the present invention can be used, for example, for developing new drugs based on protein function analysis.

Claims

 The scope of the claims

 [1] A heating / cooling element, and a heat conductor arranged in thermal contact with the heating / cooling element, and arranged in thermal contact with the heat conductor according to the temperature of the heat conductor. A temperature controller for controlling the temperature of a sample,

 Two or more heating and cooling elements are provided, and a temperature gradient is formed on a surface of the heat conductor that is in thermal contact with the sample by setting the heating and cooling elements to different temperatures. And

When the specific gravity is d [kgZm ³ ], the specific heat is cQiZ (kg'K)], and the thermal conductivity is K [WZ (m'K)], the thermal conductors are different in value expressed as dcZK. A temperature controller characterized by being composed of two or more materials!

[2] In the region where the temperature gradient of the heat conductor is formed, the average value of the dcZK value is adjusted so as to be smaller at the center of the region than at the end of the region. The temperature control device according to claim 1.

However, the average value of the dcZK value is obtained by setting the dcZK value of each material constituting the heat conductor to (dc / K) X, in a cross section perpendicular to the direction in which the temperature gradient of the heat conductor is formed. When the area occupied by the material in the above is Sx [m ² ], the value is represented by the following equation.

 [Number 2]

∑ {(d c / K) x · S x} / ∑ S x

(However, x is an integer of 1 to n, and n is the number of materials constituting the heat conductor, and is an integer of 2 or more.)

 The heating and cooling element is arranged on a lower surface of the heat conductor, the sample is arranged on an upper surface of the heat conductor,

The upper surface and the lower surface of the heat conductor are made of materials having different dcZK values from each other, and the dcZK value of the material forming the upper surface is higher than the dcZK value of the material forming the lower surface. 2. The temperature control device according to claim 1, wherein the temperature control device is adjusted to be small. [4] The heat conductor has a structure in which two layers having different dcZK values are stacked, and a layer thickness force of a layer having a small dcZK value among the two layers is a temperature gradient of the heat conductor. 2. The temperature control device according to claim 1, wherein the temperature is adjusted to be larger in a central portion of the region than in an end portion of the region where the is formed.

 [5] The heat conductor has a structure in which two or more members having different dcZK values are arranged in a direction in which a temperature gradient is formed, and the two or more members have a temperature gradient of the heat conductor. 2. The temperature control device according to claim 1, wherein the dcZK values are arranged so that the dcZK value decreases as the force increases from the end to the center of the region where the arrangement is formed.

 [6] Instead of forming the thermal conductor from two or more materials having different dcZK values,

 The heat conductor is made of at least one material, and a plurality of recesses are formed on a surface opposite to a surface that is in thermal contact with the sample, and air is present in the recesses. The temperature controller described in 1.

 7. The temperature controller according to claim 6, wherein a dimension of the concave portion in a direction in which a temperature gradient is formed is 0.5 to 20 mm.

[8] In the region where the temperature gradient of the heat conductor is formed, the depth of the concave portion is formed so as to decrease from the end to the center of the region. The temperature controller according to claim 6.

 [9] In the region where the temperature gradient of the heat conductor is formed, the number of the concave portions per unit area is formed so as to decrease from the end to the center of the region. The temperature controller according to claim 6, wherein

 [10] The temperature controller according to claim 1, wherein an interval between the heating and cooling elements is 100 mm or less.

 11. The temperature control according to claim 1, wherein a groove extending in a direction perpendicular to a direction in which a temperature gradient is formed is formed on a surface of the heat conductor that is in thermal contact with the sample. apparatus.

 12. The temperature controller according to claim 1, wherein a temperature measuring element is embedded on a surface of the heat conductor that is in thermal contact with the sample.

13. The temperature controller according to claim 1, wherein the heat conductor is made of a metal. 14. The temperature controller according to claim 1, wherein the heating / cooling element is a Peltier element.

15. The temperature control according to claim 1, wherein another heat conductor is disposed on the heat conductor, and a temperature gradient is formed on a surface of the another heat conductor that is in thermal contact with the sample. apparatus.

16. The temperature control device according to claim 15, wherein the another heat conductor has a structure in which two materials having different dcZK values are stacked.

17. The temperature control device according to claim 15, wherein the another heat conductor is two or more.

[18] The temperature measuring element includes a protection tube and a temperature sensing portion, the temperature sensing portion is disposed substantially in the center of the protection tube, the temperature gradient forming region is rectangular, and the length of the protection tube is long. When the temperature sensor is in a state where the direction and the temperature gradient forming direction are perpendicular to each other, and the temperature sensing portion is located substantially at the center of the temperature gradient forming direction in a direction perpendicular to the temperature gradient forming direction, 13. The temperature control device according to claim 12, wherein the temperature control device is embedded in the heat conductor.

[19] The temperature controller according to claim 1, further comprising another temperature measuring element for temperature monitoring, wherein the another temperature measuring element is embedded in an end of the heat conductor.

[20] The temperature controller according to claim 1, further comprising a lid disposed on the heat conductor, wherein the lid has an air layer inside.

21. The temperature adjustment device according to claim 1, further comprising a control device, wherein the output of the heating / cooling element is controlled.

22. The temperature control device according to claim 21, wherein the heating / cooling element and the heat conductor are arranged on the control device.

 23. The temperature controller according to claim 21, wherein a pair of the heating / cooling element and the heat conductor form a temperature gradient generating unit, and these are controlled by one controller.

 [24] The temperature controller according to claim 1, further comprising temperature estimating means at a predetermined location in a temperature gradient forming region of the heat conductor according to the following equation.

 Y (° C) = AX + B

 Y: temperature at a predetermined position in the temperature gradient forming region of the heat conductor

 X: Generated temperature gradient (KZmm)

 A: Distance from the center of the temperature gradient forming area to the predetermined location (mm)

B: Central temperature of the temperature gradient forming area (° C) [25] The surface of the heat conductor that is in thermal contact with the sample is covered with another heat conductor.

V. The temperature control device according to claim 1.

[26] A protein crystallization apparatus for precipitating the protein crystals from a protein-containing sample solution, wherein the temperature control apparatus according to claim 1 is used for adjusting the temperature of the sample solution. An apparatus for crystallizing a protein, comprising:

27. The protein crystallization apparatus according to claim 26, wherein the sample solution is held on a plate having a plurality of wells, and the plate is arranged so as to be in contact with a heat conductor.

28. The protein crystallization apparatus according to claim 27, further comprising a holding mechanism for holding the plate and bringing the plate into contact with the heat conductor.

[29] A mechanism in which the heat conductor is arranged with the surface in thermal contact with the plate facing downward, and the holding mechanism is arranged below the heat conductor, and moves the plate up and down. 29. The protein crystallization apparatus according to claim 28, comprising:

[30] When the heating / cooling element and the heat conductor constituting the temperature control device are a first heating / cooling element and a first heat conductor,

 Further, the second heating / cooling element was in thermal contact with the second heating / cooling element, and was thermally insulated from the first heating / cooling element and the first heat conductor. , A second heat conductor,

 28. The protein crystallization apparatus according to claim 27, wherein the second heat conductor is maintained at a constant temperature by the second heating / cooling element.

[31] The second heating / cooling element has a larger heat working surface than the first heating / cooling element, and the first heating / cooling element is provided on a heat working surface of the second heating / cooling element. Is disposed in an area of the heat acting surface which is not covered with the first heating / cooling element.

31. The protein crystallizer according to claim 30, wherein the two heat conductors are in thermal contact.

[32] The plate includes a first well and a second well having an internal space communicating with each other, the first well holds the sample solution, and the second well holds the tank. It holds a reservoir solution that does not contain impurities

31. The first well is arranged to be in thermal contact with the first thermal conductor and the second well is arranged to be in thermal contact with the second thermal conductor. The proteins described in Crystallizer.