JP2003095800A - Crystallizing tip for organic polymer and crystal forming apparatus having the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a crystallizing tip useful for crystallizing a biopolymer such as a protein and a crystal forming apparatus. SOLUTION: The crystallizing tip has a metal thin film at the interface of a substrate holding a crystallizing solution containing an organic polymer and is used for forming a crystal of the organic polymer in such a way that when the tip is contacted with the crystallizing solution, the metal thin film or an oxide thin film dissolves and the crystal of the organic polymer is formed. The crystal forming device is equipped with the crystallizing tip. It is possible to appropriately control the thickness of the thin film and the surface area contacting with the solution so as to control the dissolution amount or the dissolution speed of the metal thin film.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a crystallized chip of an organic polymer, particularly an organic polymer containing various biopolymers such as proteins and enzymes and a complex thereof, and a crystal production apparatus having the crystallized chip.

[0002]

2. Description of the Related Art Generally, crystal structure analysis is important for understanding the functions of biopolymers such as proteins. In particular, the crystal structure analysis of a biopolymer using X-ray diffraction is extremely useful because it can grasp the entire atomic arrangement. However, for that purpose, it is necessary to prepare a crystal satisfying the conditions (for example, crystallinity / size) necessary for crystal structure analysis by X-ray diffraction.

Heretofore, the following method has been conventionally used as a means for crystallizing biopolymers such as proteins.

(1) A batch method for obtaining crystals by mixing a protein solution and a precipitant solution in a supersaturated environment and allowing them to stand.

(2) A vapor diffusion method in which the crystallization solution is concentrated by utilizing the saturated vapor pressure difference between the two liquids and the concentration of the protein solution and the concentration of the precipitant are increased to promote crystal nucleation and further crystal growth.

(3) A liquid phase diffusion method for promoting crystal nucleation and crystal growth by diffusing and mixing a protein and a precipitant by contacting two liquids having different concentrations.

(4) A dialysis method which promotes crystal nucleus growth and crystal growth by bringing two proteins having different protein concentrations into contact with each other through a semipermeable membrane to diffuse and mix the protein and the precipitant.

Among these crystallization methods, the vapor diffusion method widely used for protein crystallization is the two solutions having different precipitant concentrations, that is, a crystallization solution having a high precipitant solution concentration and a low precipitant solution concentration. Crystallization is promoted by the combined action of concentrating the crystallization solution using the saturated vapor pressure difference between the solutions and decreasing the solubility by increasing the concentration of the precipitating agent.

This vapor diffusion method is advantageous over other methods in that it can be crystallized with a small amount of protein solution,
Since crystallization conditions (for example, pH, protein concentration, type and concentration of precipitating agent, temperature, etc.) need to be determined by trial and error, crystallization of proteins requires considerable effort, There are many cases in which we cannot find it.

As a tool used for optimizing these crystallization conditions, a crystallization screening kit and the like are commercially available. This is the condition of reagents with high crystallization success rate among known protein crystallization conditions (for example, reagent type, concentration,
pH and the like) are set in advance and are widely used to search for appropriate crystallization conditions. For example, in the crystal screen (trade name) of Hampton Co. in the United States, 48 kinds of precipitants (27 kinds of inorganic salts,
Reagent conditions consisting of 98 types of matrices by combining 10 types of organic solvents, 13 types of polymers) and 10 types of buffers are provided, and it is widely used as a primary screening means in crystallization of proteins whose crystallization conditions are unknown. It's being used.

For certain proteins, good quality crystals are obtained when a precipitating agent which is usually used, for example, an inorganic salt such as sodium chloride or ammonium sulfate, an organic solvent such as MPD, and a water-soluble polymer such as polyethylene glycol is used alone. May not be obtained and crystallization may require specific ions in addition to these precipitants. In the crystallization screening kit described above, these specific ions are also incorporated as conditions, but due to the limitation of the number of conditions, they are incorporated in comparison with solution parameters (precipitant conditions, buffer conditions, pH, etc.). There are few conditions. Therefore, the screening kit cannot completely cover the condition matrix for a specific ion.

That is, the effect of the added element on the crystallization of the protein cannot be investigated with the current screening kit. On the other hand, in order to investigate the effect of these additional elements, it is necessary to conduct a large number of experiments to investigate the crystallization feasibility of proteins whose crystallization conditions are unknown under the conditions of all ions. Inefficient and impractical.

On the other hand, in the crystallization of protein, it is necessary to use the mass transfer in the crystallization solution to shift from the unsaturated state to the supersaturated state, but in order to cause spontaneous crystal nucleation, the unsaturated state is generated. It is necessary to increase the supersaturation above the solubility curve which is the boundary between the supersaturated state. For example, protein crystallization generally requires a protein concentration of several times to several tens of times the solubility. In other words, a relatively high degree of supersaturation is required for protein crystal nucleation. On the other hand, since the frequency of crystal defects, which is directly related to the crystal quality, increases with the crystal growth rate, and the crystal growth rate increases with the supersaturation degree, the supersaturation degree is required to obtain a good quality crystal suitable for crystal structure analysis. It is desirable to grow crystals in a low state. That is, the optimal conditions for the degree of supersaturation differ in both protein nucleation and crystal growth processes. The existing crystallization means described above cannot realize the optimum conditions for both processes in the same experiment.

As a means for realizing the optimum conditions of both of these processes, there is a crystal production apparatus which utilizes electrostatic interaction between protein charges and charges of the crystallization substrate for crystal nucleation (Japanese Patent Laid-Open No. 8-294601). . This utilizes the fact that the dissociation state of amino acid residues and the surface state of the crystallization substrate depend on the solution pH. However, the above device has the following problems.

(1) In the vicinity of the point where the total charge of the protein is zero (isoelectric point), since there is no interaction between the protein molecule and the base material, it does not contribute to the formation of crystal nuclei.

(2) Crystallization may not proceed because the electrostatic interaction between the protein and the substrate is too strong and the protein molecules important for crystallization do not rearrange.

(3) In a high ionic strength environment, which is a general crystallization environment, the effect is low in some cases because the interaction is at an extremely short distance due to the compression of the diffusion layer in the vicinity of the protein molecule and the substrate. is there.

(4) p where the crystallization substrate is ideally charged
Not applicable when the structure of the protein molecule is unstable in H.

(5) p where the crystallization substrate is ideally charged
In H, protein crystallization may not occur.

[0020]

As a means for realizing nucleation and growth under the same experiment, a method of locally controlling the concentration of these metal ions in protein crystallization can be considered. However, local control of these metal ion concentrations in protein crystallization cannot be realized by a combination of conventional crystallization techniques and solution addition.

For example, in the vapor diffusion method, the metal ion concentration in the crystallization solution depends on the concentration of the crystallization solution as the vapor diffusion progresses, so that the metal ion concentration is the same as that of other reagents contained in the crystallization solution. It exhibits concentration behavior and cannot independently control the metal ion concentration. Further, since the concentrated layer is formed in the vicinity of the gas-liquid interface, water in the obtained crystals evaporates, and there is a high risk of so-called dry-up in which protein crystals collapse.

In the liquid phase diffusion layer, a concentrated layer is formed by the diffusion of metal ions in the liquid phase, but since the speed thereof substantially depends only on the diffusion constant of the metal ion in the solution, the concentrated layer can be formed freely. There is no degree.

A method of sequentially adding a metal ion-containing solution to a crystallization solution in a vapor diffusion method or a batch method is also conceivable, but it has the same problems as in the liquid phase diffusion method, and the experimental equipment becomes complicated, which makes it more versatile. Lack.

As described above, with the conventional crystallization means, it is difficult to obtain the crystallization conditions that cover all the conditions, and a great deal of labor is required. Therefore, the crystal nucleation and the crystal nucleation under different optimal conditions are performed. Both processes cannot be realized in a wide experimental range and in the same experiment. An object of the present invention is to provide a crystallizing chip of an organic polymer capable of solving such a problem and covering a wide range of crystallizing conditions, and a crystal producing apparatus equipped with the crystallizing chip.

[0025]

According to the present invention, a metal thin film is provided on at least a part of a substrate holding a crystallization solution containing an organic polymer, and a metal is dissolved from the metal thin film by contact with the crystallization solution. Then, the crystallization chip is characterized by producing crystals of an organic polymer.

The crystallization chip of the present invention is characterized by having means for controlling the amount or rate of dissolution of the metal thin film. The amount or rate of dissolution is controlled by the thickness of the metal thin film or the contact area with the crystallization solution.
The amount or rate of dissolution can be controlled by the composition of the metal thin film. Further, the dissolution amount or dissolution rate is controlled by the arrangement of the second thin film layer formed on the metal thin film.

In the present invention, the metal thin film is preferably composed of a metal simple substance, an alloy or a metal oxide thin film. Further, in the present invention, the organic polymer is preferably a protein.

Still another aspect of the present invention has a metal thin film at the interface of a base material holding a crystallization solution containing an organic polymer, and the metal is dissolved from the metal thin film by contact with the crystallization solution, It is a crystal production apparatus provided with a crystallization chip for producing the crystal of.

[0029]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention has a metal thin film on at least a part of a substrate holding a crystallization solution containing an organic polymer, and the metal thin film and / or the oxide thin film is formed by contact with the crystallization solution. A crystallization chip characterized by being dissolved to generate crystal nuclei of an organic polymer and grow crystals.

In the present invention, as the base material for holding the crystallization solution, not only inorganic materials such as silicon, glass, quartz, metal and ceramics but also resin materials such as polystyrene, polypropylene and polyethylene can be preferably used, but it is not particularly limited. There is no. The shape and size of the base material can be appropriately adjusted according to the operability of the experiment.

Next, the metal thin film is not particularly limited as long as it is a material that can be a source of metal ion dissolution, and a metal simple substance,
Materials such as alloys or metal oxides are used. In particular, metals that may have specific interactions with proteins, such as magnesium (Mg), calcium (Ca), manganese (Mn), iron (Fe), cobalt (Co), nickel (Ni), copper (Cu). Metals that become divalent ions, such as zinc, zinc (Zn), cadmium (Cd), and cerium (Ce), alloys containing these metals, and oxides of these metals can be effectively used. The alloy composition is appropriately adjusted in order to adjust the dissolution amount and dissolution rate of metal ions.

As a method for forming a metal thin film on a substrate, a dry process such as vapor deposition or sputtering, or a wet process such as electroplating or electroless plating is usually used, and the method is selected according to a desired thin film thickness or thin film area. Is appropriately selected.

The shape of the metal thin film can be arbitrarily adjusted for the purpose of controlling the amount and rate of dissolution as described above. For example, reducing the area of the metal thin film to suppress the dissolution rate,
The amount of dissolution can be suppressed by reducing the area or the thickness of the metal thin film.

In order to control the dissolution rate of the metal thin film, a second thin film layer can be partially formed on the metal thin film. By partially forming the second thin film layer on the metal thin film, it inhibits the supply of ions from the crystallization solution necessary to react with the metal thin film and dissolve the metal ions and the diffusion path of the generated metal ions. To do. Further, the elution rate of the electrochemically base metal can be increased by the metal material of the second film layer electrically connected to the metal thin film.

Further, in order to realize a plurality of crystallization conditions in the same container, a plurality of metal thin films can be installed on the base material. In this case, various metal ion effects are investigated in the same crystallization chip, so that the shape and material of the metal thin film can be made different. Further, in order to investigate the concentration dependence of various metal ion effects and the dissolution rate dependence of metal ions in the same crystallization chip, the film thickness and film area of the metal thin film can be made different.

Further, by integrating the crystallization chip of the present invention on a large scale and further combining it with an automatic crystallization solution setting device, an automatic crystallization state observation device, etc., the crystallization experiment conditions can be efficiently searched. can do. In this case, the crystallization experiment layout can be simplified as compared with the conventional method by utilizing the fact that the solubility of the protein is significantly changed by the metal ion.

Next, when the crystallization chip of the present invention is applied to a crystal production apparatus of the vapor diffusion method, the crystallization chip can hold the droplet shape expected by a part of the crystallization solution having a low surface tension. It is also possible to provide a means for holding the chemical solution.
For example, the droplet holding portion is bonded or adhered to the surface edge portion on the base material, or an uneven droplet holding portion is provided on the surface of the base material. Further, means such as coating or chemical modification on the surface of the base material, particularly on the surface edge of the base material to increase water repellency can be used.

The crystallization chip of the present invention can be applied to various organic polymer crystallization methods such as a microbatch method, a vapor diffusion method, a liquid phase diffusion method and a dialysis method. The crystallization conditions in these means can be set arbitrarily. In that case, the crystallization conditions selected from the conventional screening kit can be applied. In this case, a metal salt precipitate may be generated due to the coexistence of the metal ion and the specific anion, but the generation of the metal salt is controlled or avoided by appropriately selecting the combination of the types of the metal ion and the anion. be able to. When the metal thin film is not sufficiently dissolved, the elution of metal ions from the metal thin film can be accelerated by mixing the crystallization solution with a specific compound that promotes metal dissolution.

Next, according to the present invention, a metal thin film is provided on at least a part of a base material holding a crystallization solution containing an organic polymer, and the metal is dissolved from the metal thin film by contact with the crystallization solution, It is a crystal production apparatus provided with a crystallization chip for producing polymer crystals. That is, the crystal production apparatus of the present invention comprises the above-mentioned crystallization chip,
For example, the microbatch method, vapor diffusion method, liquid phase diffusion method, dialysis method and the like are incorporated into various organic polymer crystallization production apparatuses.

The crystallization chip and the crystal growth apparatus of the present invention will be described below with reference to the drawings. FIG. 1 shows a schematic view of a crystallization chip T ₁ used in the crystal growth apparatus of the present invention. In the figure, the crystallized chip T ₁ has a metal thin film 2 formed on the interface of a base material 1. The base material 1 is made of metal, ceramics, glass, quartz, silicon or resin material. A metal thin film 2 is formed on the surface of the base material 1 by a method such as vapor deposition or sputtering. Here, the thickness of the metal thin film is usually in the range of 1 nm to 10 μm.

Here, the principle of crystal formation of an organic polymer using the crystallization chip of the present invention will be described based on FIG. 1 (c). The mechanism of crystal nucleation of the organic polymer is based on the supply of metal ions by the dissolution reaction of the metal thin film 2 in the crystallization solution. The supply of the metal ions is performed in the following (a) to (d)
It consists of the process of. (A) The ions P1 necessary for the dissolution reaction diffuse in the crystallization solution 3 in the direction of the metal thin film 2. (B) Ions P1 necessary for the dissolution reaction are adsorbed on the surface of the metal thin film 2. (C) The ion P1 causes a dissolution reaction on the surface of the metal thin film 2. (D) The eluted metal ions P2 are desorbed from the metal thin film 2. (E) The generated metal ions P2 diffuse into the crystallization solution 3 to form crystal nuclei.

For the above steps (a) to (d), (e)
If the diffusion of the metal ions in the process is sufficiently slow, the metal ions stay in the vicinity of the interface with the progress of the dissolution reaction and the concentration increases. That is, a local increase in metal ion concentration, which cannot be achieved by the conventional crystallization method, is realized.
Due to this local increase in the concentration of metal ions, the degree of supersaturation in the vicinity of the metal thin film is increased as compared with the entire solution, which promotes crystal nucleation. On the other hand, since the degree of supersaturation of the entire solution gradually increases as the diffusion of metal ions progresses, the generated crystal nuclei can grow under a lower saturation condition. That is, the most important optimum conditions for nucleation and crystal growth can be easily realized with the same crystal production apparatus.

The types of these metal ions, the amount of dissolution,
The dissolution rate can be controlled by the above-mentioned thin film shape (film composition / film thickness / area), the second thin film layer formed on the metal thin film, and the like. Further, the type of metal ions to be eluted can be arbitrarily selected according to the composition of the metal thin film, and the amount of dissolution can be controlled with extremely high precision by using the film composition, film thickness, area and the like. Further, the dissolution rate can be controlled by changing the area of contact with the crystallization solution by adjusting the area of the metal thin film. Furthermore, the dissolution reaction rate can be controlled by adding an element that changes the solubility (corrosion resistance) of the metal thin film. Further, as described above, it is possible to control the dissolution rate of metal ions by the second thin film layer formed on the metal thin film.

Next, FIG. 2 shows another embodiment of the crystallization chip used in the crystal producing apparatus of the present invention. On the upper side of the metal thin film 22 formed on the base material 21, a second thin film layer 23 for covering almost the entire metal thin film 22 and adjusting the solubility thereof is formed. By forming the second thin film layer 23, the supply of ions necessary for the metal ion dissolution reaction to the metal thin film 22 and the diffusion of the eluted metal ions are suppressed. Further, the metal, which is the material of the second thin film layer electrically connected to the metal thin film 22, increases the elution rate of the electrochemically base metal, thereby facilitating or suppressing the metal ions. The material of the second thin film layer may be metal, ceramic, glass or resin material, and its thickness is usually 1
It is set in the range of nm to 1 μm.

Next, the second thin film layer 3 of the crystallized chip of the present invention
As shown in FIG. 3, 3 is formed with a smaller size on the upper side of the metal thin film 32, and the upper surface of the metal thin film 32 is partially exposed. Therefore, since it has a smaller contact area with the crystallization solution, elution of metal ions is suppressed.

The crystallized chip shown in FIG. 4 has a second thin film layer 43 formed on a metal thin film 42 formed on a base material 41 so as to partially overlap therewith. Here, the metal material of the metal thin film 42 and the second thin film layer 43 can be appropriately selected, the dissolution rate of each metal ion can be adjusted, and the overall dissolution rate can be adjusted appropriately.

The crystallized chip shown in FIG. 5 is one in which a metal thin film 52 and a second thin film layer 53 are juxtaposed on a base material 51. In this embodiment, the metal thin film 52 and the second thin film layer 5 are
By selecting the metal, alloy, or metal oxide material of No. 3, the dissolution rate of the metal ions in the respective metal thin film 52 and second thin film layer 53 can be adjusted. As a result, the dissolution amount and dissolution rate of the metal ion as a whole can be adjusted with higher accuracy.

In the crystallization chip shown in FIG. 6, four metal thin films (62a, 62b, 62c, 62d) are arranged on a base material 61. The shape and dimensions of these metal thin films are substantially the same, but the materials of the four metal thin films are all different. In this case, different dissolution rates of different metal ion species will be obtained at the interfaces of the respective thin films.

The crystallization chip shown in FIG. 7 has four kinds of metal thin films (72a, 7a) having different shapes and sizes on a base material 71.
2b, 72c, 72d) are formed. Further, these metal thin films (72a, 72b, 72c, 72d) are made of different materials. As a result, the surface of each of the metal thin films 72a to 72d has different dissolution rates due to different metal ions, and a crystallization chip having various crystallization conditions can be obtained.

The crystallization chip shown in FIG. 8 has four different types of metal thin films (82a, 82b, 8) on a base material 81.
2c, 82d) are arranged. The materials of these metal thin films are the same. Each surface exhibits different dissolution rates of metal ions.

The crystallization chip shown in FIG. 9 has four metal thin films arranged in four rows and six metal rows arranged on a base material 91. The metal thin films in the columns all have the same shape and use different materials. On the other hand, the metal thin films (91a to 92f) in the rows have different shapes and use the same material. As a result, the base material 91 has 24 different crystallization conditions.

In the crystallization chip shown in FIG. 10, a metal thin film 102 is arranged on the central upper surface of a base material 101, and a droplet holding portion 103 having a predetermined width and a predetermined thickness is provided on the surface edge portion thereof. . The droplet holding unit 103 drops the crystallization solution onto the upper surface of the metal thin film and crystallizes the solution, so that the droplet is formed into the base material 1.
01 to prevent leakage. Here, the droplet holding unit 103
The surface can be treated to be water repellent in order to further enhance its effect.

In the crystallized chip shown in FIG. 11, a metal thin film 112 is arranged on the central upper surface of a base material 111 whose central portion is dug to a predetermined depth. Then, the height and width of the droplet holding portion 113 can be appropriately adjusted according to the amount of the crystallization solution.

In the crystallization chip shown in FIG. 12, a metal thin film 122 is arranged on the central upper surface of a base material 121, and a droplet holding member 123 is provided on the edge of the surface. Here, the droplet holding member 123 is formed of a coating or a sheet for water-repellent treatment of its surface. Therefore, even if the height of the droplet holding member 123 is reduced, it is possible to effectively prevent the droplets from leaking from the base material.

Next, a crystal production apparatus using the crystallization chip of the present invention will be described. Figure 13 is provided with a crystallization chip T ₁₃ of the present invention, showing a crystal formation apparatus of the so-called micro-batch process. The crystallization chip T ₁₃ is placed in the crystallization solution 133 in which the protein and the precipitant solution are mixed so as to be in a supersaturated state, and the metal nuclei 134 are generated by the elution of the metal ions from the metal thin film 132 to promote the crystal growth. be able to.

[0056] Figure 14 shows the crystal generator of vapor diffusion method by hanging drop having a crystallization chip T ₁₄ of the present invention. The crystallization solution is concentrated by utilizing the saturated vapor pressure difference between the crystallization solution droplet 143 and the reservoir liquid 145 to increase the protein concentration and the precipitant concentration.
Production of 44 and crystal growth can be promoted. The 15 with a crystallization chip T ₁₅ of the present invention, showing a crystal formation apparatus of the vapor diffusion method by sitting drop method.
The principle is the same as in FIG.

[0057] Figure 16 shows a crystal generator of the liquid phase diffusion method with a crystallization chip T ₁₆ of the present invention. When the two droplets 163 and 165 having different protein concentrations are brought into contact with each other, the protein and the precipitant diffuse and mix, and the crystal nucleus 16
4 can be generated to promote crystal growth.

[0058] Next Figure 17 shows a crystal generator according dialysis method with a crystallization chip T ₁₇ of the present invention. Semipermeable membrane 175
Crystallization solutions 173 and 176 having different concentrations via
The crystallization solution of the protein and the precipitating agent diffuses and mixes by contacting the crystal nuclei 17
4 is generated to grow crystals.

[0059]

EXAMPLES In order to verify the effect of the crystal growth apparatus of the present invention, comparison was made with the conventional crystallization technique under the crystallization conditions shown in Table 1. This condition is a crystallization condition in which the concentration of catalase after the concentration is completed is equal to or lower than the solubility of catalase in the coexistence of cobalt, and crystals are not usually obtained by the vapor diffusion method. The experimental results obtained are shown in Table 2.

As is clear from Table 2, crystal precipitation was observed in Example 1 in which a cobalt thin film was formed on a silicon substrate and in Example 2 in which a cobalt thin film was formed on a glass substrate.

[0061]

[Table 1]

[0062]

[Table 2]

The embodiments and examples disclosed this time are to be considered as illustrative in all points and not restrictive. The scope of the present invention is shown not by the above description but by the claims, and is intended to include meanings equivalent to the claims and all modifications within the scope.

[0064]

EFFECTS OF THE INVENTION By using the crystallization chip and the crystal production apparatus of the present invention, both nucleation and crystallization growth processes under different optimum conditions in crystallization of organic polymers such as proteins can be conducted over a wide range of experimental ranges. And it becomes possible to realize it in the same experiment. This can be expected to improve the crystallization and crystallinity of proteins that are difficult to obtain by existing crystallization methods. In the field of structural genomics, it is necessary to crystallize proteins whose crystallization conditions are unknown,
By using the crystallization chip and the crystal production apparatus of the present invention, the crystallization success rate can be expected to improve. On the other hand, when the technique of adding metal ions is used together, the concentration of protein that can be crystallized can be reduced, and as a result, the amount of protein that is required to search for crystallization conditions can be reduced. This is extremely useful in increasing the number of experimental conditions, especially when structural analysis of a protein having a low expression level is required. In addition, the effect of specific metal ions that cannot be verified by the existing screening matrix can be extensively tested only by changing the experimental base material, which enables efficient search for crystallization conditions.

[Brief description of drawings]

FIG. 1A is a perspective view of a crystallization chip of the present invention. (B) is a front view of the crystallized chip. (C) is a mechanism diagram for forming crystal nuclei around the crystallization chip.

FIG. 2A is a perspective view of a crystallization chip of the present invention. (B) is a front view of the crystallized chip.

FIG. 3A is a perspective view of a crystallization chip of the present invention. (B) is a front view of the crystallized chip.

FIG. 4A is a perspective view of a crystallization chip of the present invention. (B) is a front view of the crystallized chip.

FIG. 5A is a perspective view of a crystallization chip of the present invention. (B) is a front view of the crystallized chip.

FIG. 6A is a perspective view of a crystallization chip of the present invention. (B) is a front view of the crystallized chip.

FIG. 7A is a perspective view of a crystallization chip of the present invention. (B) is a front view of the crystallized chip.

FIG. 8A is a perspective view of a crystallization chip of the present invention. (B) is a front view of the crystallized chip.

FIG. 9A is a perspective view of a crystallization chip of the present invention. (B) is a front view of the crystallized chip.

FIG. 10A is a perspective view of a crystallization chip of the present invention. (B) is an AA sectional view of the crystallization tip of (a).

FIG. 11A is a perspective view of a crystallization chip of the present invention. (B) is an AA sectional view of the crystallization tip of (a).

FIG. 12 (a) is a perspective view of a crystallization chip of the present invention. (B) is an AA sectional view of the crystallization tip of (a).

FIG. 13 is a schematic view of a crystal growth apparatus of the present invention by a microbatch method.

FIG. 14 is a schematic view of a crystal growth apparatus of the present invention by a vapor diffusion method (hanging drop).

FIG. 15 is a schematic view of a crystal growth apparatus of the present invention by a vapor diffusion method (sitting drop).

FIG. 16 is a schematic view of a crystal growth apparatus of the present invention by a liquid phase diffusion method.

FIG. 17 is a schematic view of a crystal growth apparatus of the present invention by a dialysis method.

[Explanation of symbols]

T ₁ crystallization chip, 1 base material, 2 metal thin film, 3 crystallization solution, P 1 ion, P 2 metal ion, 21, 3
1, 41, 51, 61, 71, 81, 91, 101, 1
11,121 Base material, 22, 32, 42, 52, 62
a, 72a, 82a, 92a, 102, 112, 122
Metal thin film, 23, 33, 43, 53 Second thin film layer.

Claims (8)

[Claims] 1. A metal thin film is provided on at least a part of a base material holding a crystallization solution containing an organic polymer, and a metal is dissolved from the metal thin film by contact with the crystallization solution,
A crystallization chip characterized by producing crystals of an organic polymer. 2. The crystallized chip according to claim 1, further comprising means for controlling a dissolution amount or a dissolution rate of the metal thin film. 3. The crystallization chip according to claim 1, wherein the dissolution amount or the dissolution rate is controlled by the film thickness of the metal thin film or the contact area with the crystallization solution. 4. The crystallized chip according to claim 1, wherein the amount or rate of dissolution is controlled by the composition of the metal thin film. 5. The crystallized chip according to claim 1, wherein the dissolution amount or the dissolution rate is controlled by the second thin film layer formed on the metal thin film. 6. The crystallized chip according to claim 1, wherein the metal thin film is a metal simple substance, an alloy or a metal oxide thin film. 7. The organic polymer is a protein.
The crystallization chip described. 8. A crystal production apparatus comprising the crystallization chip according to claim 1.