JP4865680B2 - Protein crystallizer - Google Patents

Description

  The present invention relates to a protein crystallization apparatus.

A vapor diffusion method is known as a method for searching for protein crystallization conditions. And the apparatus which automates a vapor diffusion method is also considered, and the technique of patent document 1 is known as this kind of apparatus, for example.
In the technique described in Patent Document 1, a sitting drop method is used, a transporting means for a crystallization container (crystallization plate), a storage section for storing a plurality of crystallization containers and chip racks, and It is configured to include transport means for transporting these dispensing materials, and it is possible to successively produce crystallization plates subjected to a desired crystallization dispensing process.
Japanese Patent No. 3753134

  However, in the technique described in Patent Document 1, although only the sitting drop method is used as the vapor diffusion method, for example, in order to produce only crystallization plates of the same specification one after another, the production efficiency is excellent. For example, when changing the vapor diffusion method to another method (for example, the hanging drop method) and appropriately changing them, or when it is necessary to produce a small number of crystallization plates with different specifications, On the other hand, it is inconvenient for requests.

Furthermore, in the technique described in Patent Document 1, a desired dispensing process is continuously performed on the crystallization plate so that crystallization plates having the same specifications can be manufactured one after another. There is also a problem that a conveying means or the like is required, and the apparatus becomes large and the cost is high.
Accordingly, the present invention has been made paying attention to such problems, and an object thereof is to provide a protein crystallization apparatus that is small and easy to use.

  In order to solve the above problems, the present invention provides a protein crystallization apparatus, a housing having a processing chamber for performing a desired dispensing operation on a crystallization plate, and the processing provided in the housing. A base that is slidably movable between a working position located in a room and an overhanging position that projects outside the processing chamber, and provided on the base to seal the crystallization plate and a well opening of the crystallization plate A sealing device that can be sealed using a member, a dispensing processing unit in which a dispensing material necessary for the dispensing operation is disposed, and an upper part of the dispensing processing unit in the processing chamber, Dispensing head driving device having a dispensing head for performing necessary processing such as dispensing to the well of the crystallization plate and capable of moving the dispensing head to a position necessary for processing to the well of the crystallization plate When, An electrical component chamber having a control device for controlling the dispensing head driving device and the sealing device, respectively, and an unnecessary material collecting unit capable of collecting unnecessary materials generated by the dispensing operation, provided below the base. It is characterized by having.

  The protein crystallization apparatus according to the present invention has a processing chamber in its housing, and a base on which a dispensing processing unit including a sealing device is provided is disposed in the processing chamber. Since the working position and the overhanging position that protrudes outside the processing chamber are slidably movable, if the base is placed in the overhanging position, the dispensing processing unit at the top is moved forward. It can be pulled out. For this reason, for example, necessary operations such as replacement of necessary dispensing materials and crystallization plates can be performed outside the processing chamber. Therefore, since the work space for manpower in the processing chamber itself can be reduced within a necessary and sufficient range, the apparatus can be miniaturized.

  In other words, for example, even if the processing chamber is configured to be sufficiently small, the base can be positioned at the overhanging position and the dispensing processing unit at the top can be pulled out forward. Therefore, handling and maintenance before and after dispensing work are easy. Therefore, for example, when a plurality of methods are applied as the vapor diffusion method, for example, even when the sitting drop method and the hanging drop method are appropriately changed and used, each dispensing operation arranged in the dispensing processing unit Therefore, it is possible to easily replace the necessary dispensing material for the crystallization plate corresponding to each dispensing operation. Therefore, for example, when a plurality of vapor diffusion methods are appropriately changed and used, or when it is necessary to produce a small number of crystallization plates having different specifications, the convenience of the so-called multi-product small-lot requirements is good. .

Furthermore, according to the protein crystallization apparatus of the present invention, the casing is provided with the above-described dispensing head driving device, the base having the dispensing processing unit on the upper surface, and the control device in the electrical component chamber below the base. Since the unnecessary material recovery unit is sequentially installed from the top of the housing, this arrangement contributes to the realization of a compact configuration, and is suitable as a configuration for downsizing the apparatus.
Here, in the protein crystallization apparatus according to the present invention, the processing chamber has an opening / closing door formed of a material that can visually recognize the processing chamber, and the base is formed from a portion where the opening / closing door is opened. It is preferable to be able to be positioned at the projecting position by projecting out of the processing chamber. With such a configuration, since the open / close door is formed of a material that allows the inside of the processing chamber to be visually recognized, the state of the processing chamber can be easily confirmed. And since the base can be positioned at the extended position by projecting from the opened part of the door to the outside of the processing chamber, it is more suitable as a configuration that can downsize the apparatus and improve its usability. It is.

By the way, this type of “protein crystallization apparatus” is usually used at a low temperature (for example, 4 ° C.) because the protein solution is denatured at room temperature. However, performing dispensing work for a long time in a low temperature room places a heavy burden on the operator.
Therefore, in the protein crystallization apparatus according to the present invention, it is preferable to perform dispensing by installing the entire protein crystallization apparatus in a cool box. Such a configuration is suitable for preventing denaturation of the protein solution and improving the crystallization quality. That is, since the protein crystallization apparatus according to the present invention has a compact configuration as described above, the entire protein crystallization apparatus can be easily placed in a research refrigerator such as a chromatography chamber (registered trademark). It is suitable for reducing the burden on the operator.

Furthermore, in the protein crystallization apparatus according to the present invention, when the entire protein crystallization apparatus is installed in a cool box and dispensed, the set temperature of the cool box is set to 4 ° C. It is preferable to perform the dispensing operation at temperature. Such a configuration is more suitable for improving the quality of crystallization.
Moreover, in the protein crystallization apparatus according to the present invention, it is preferable that the control device is surrounded by a heat insulating material. Such a configuration is suitable for preventing condensation of the electrical components by preventing the electrical components in the control device from being cooled to a low temperature.

In the protein crystallization apparatus according to the present invention, it is preferable that the electrical component chamber is surrounded by a soundproofing material. Such a configuration is suitable for reducing noise.
In the protein crystallization apparatus according to the present invention, it is preferable that the electrical component chamber and the processing chamber are partitioned by a heat insulating material. Such a configuration is suitable for keeping the temperature in the processing chamber at the set temperature of the cold storage when the entire protein crystallization apparatus is installed in the cold storage and dispensing is performed.

  As described above, according to the present invention, a protein crystallization apparatus that is small and easy to use can be provided.

Hereinafter, an embodiment of a protein crystallization apparatus according to the present invention will be described with reference to the drawings as appropriate. 1 is an external view of an embodiment of the protein crystallization apparatus according to the present invention as viewed from the front, and FIG. 2 (a) is an exterior cover including the opening / closing door in FIG. The figure of a state is shown and FIG.2 (b) is A arrow directional view in Fig.2 (a).
As shown in FIG. 1, the protein crystallization apparatus 1 includes a substantially rectangular parallelepiped housing 2 whose longitudinal direction is up and down, and has a processing chamber 4 in the center in the vertical direction. In addition, an equipment chamber 3 is provided in the upper portion of the processing chamber 4, and an electrical component chamber 5 is provided in the lower portion of the processing chamber 4. The electrical component chamber 5 and the processing chamber 4 are separated from each other by a heat insulating material 5a. The protein crystallization apparatus 1 is used for dispensing the entire protein crystallization apparatus 1 in a research cool box (not shown) such as a chromatographic chamber (registered trademark). The set temperature of the storage is set to 4 ° C. Then, the dispensing operation is performed with the inside of the processing chamber 4 set at the set temperature.

  Here, the processing chamber 4 located in the center is a chamber for performing a desired dispensing operation on the crystallization plate, and prevents the intrusion of dust into the inside and manages the internal atmosphere in a necessary state. The periphery is covered with an exterior cover 3a as possible. An opening / closing door 7 is provided in front of the processing chamber 4. The open / close door 7 is a double door. In other words, the open / close door 7 has left and right open / close doors 7a and 7b connected to the housing 2 via hinges 7c provided at the left and right ends, respectively, whereby the hinge 7c is used as a support shaft. It can be opened and closed. Then, by opening the door, it is possible to form an opening having a width capable of performing necessary work including taking in and out of the dispensing material. Here, the open / close door 7 is formed of a transparent tempered glass material in which the open / close doors 7a and 7b can visually recognize the inside of the processing chamber 4 from the outside.

As shown in FIG. 2, a base 8 is provided in the processing chamber 4 on the processing chamber bottom 4a. The base 8 includes a base drawer mechanism 22 at a lower portion thereof, and is provided on the bottom 4a of the processing chamber through the base drawer mechanism 22 so as to be slidable.
In detail, the base drawer mechanism 22 is provided on the upper surface of the processing chamber bottom 4a on the fixed guide rails 22a on the two right and left sides extending along the paper surface direction in the drawing, and on each fixed guide rail 22a. A movable side guide rail 22b that is supported and is slidable along the fixed side guide rail 22a. The base body 8a having a rectangular shape in plan view is formed on the movable side guide rail 22b. Is fixed. In addition, gripping handles 8b are attached to the base body 8a at two locations on the front surface thereof. The base body 8a is disposed at a position facing the opening when the opening / closing door 7 is opened, and the base body 8a is pushed and pulled by the hand to pull the base body 8a to the guide rail 22a. The base body 8a of the base 8 is slidable in the direction opposite to the position where the open / close door 7 is opened.

  As a result, the base body 8a is pulled out to the front side of the paper surface in the drawing, and is positioned at the projecting position where the base body 8a projects from the opening of the open / close door 7 to the outside of the processing chamber 4 and the work position positioned within the processing chamber 4. It is supposed to be. Here, the top of the base body 8a is a dispensing processing unit 50 for performing a desired dispensing operation on the crystallization plate. The dispensing processing unit 50 is configured so that the entire dispensing processing unit 50 can be drawn out of the processing chamber 4 by the base drawing mechanism 22 so that dispensing materials and the like can be easily taken in and out. Has been.

  Hereinafter, the dispensing processing unit 50 will be described with reference to FIGS. 3 and 4 as appropriate. Here, in the protein crystallization apparatus 1, the dispensing processing unit 50 is configured so that both the sitting drop method and the hanging drop method, which are known as vapor diffusion methods, can be used. Thereby, according to this protein crystallization apparatus 1, the crystallization plate according to the crystallization method to be utilized can be installed, and the crystallization plate can be replaced according to the crystallization method. ing. 3 and 4 are plan views of the dispensing processing unit 50, FIG. 3 is a layout diagram when using the sitting drop method, and FIG. 4 is a layout diagram when using the hanging drop method. .

As shown in FIGS. 3 to 4, the dispensing processing unit 50 includes a crystallization plate, a sealing device that can seal the well opening of the crystallization plate using a sealing member, and a crystallization dispensing unit. Dispensing materials and the like necessary for processing are arranged.
Specifically, in the dispensing processing unit 50, a large chip rack 51 and a small chip rack 52 are respectively installed from the left side on the back side in a plan view. The large tip rack 51 is provided with a plurality of large tips (for large volumes) for dispensing reservoir solutions, and the small tip rack 52 is provided with small tips for dispensing droplets (small volumes). For use). Both chips are disposable (disposable) type, and are discarded after use to prevent contamination between the solutions.

  Therefore, for disposal, the dispensing processing unit 50 is provided with a trash can 9 (see FIG. 1 to FIG. 1) installed below the dispensing processing unit 50 at a position between the large chip rack 51 and the small chip rack 52. 2), a trash box entrance 53 communicating with the trash box 9 is provided, and used chips can be thrown into the trash box 9 from the trash box entrance 53. The used chips accumulated in the trash box 9 can be emptied by pulling out the trash box 9 from the front surface of the protein crystallization apparatus 1 (see FIGS. 1 and 2).

  Further, as shown in FIGS. 3 to 4, a reservoir container 54 containing a reservoir solution is installed on the front side of the large chip rack 51. A plurality of deep wells are formed in the reservoir container 54, and a plurality of reservoir solutions for searching for crystallization conditions are stored in the deep wells. The reservoir container 54 is of a different type from the plurality of reservoir solutions stored therein. The reservoir container 54 is equipped with a lid 54a that can be opened and closed by driving a rotary air cylinder 54b for the purpose of preventing drying of the reservoir solution. The air cylinder 54b is appropriately driven according to a signal from the control device 20, so that the opening / closing operation of the lid 54a is appropriately executed.

  A protein tube 55 containing a protein solution is installed on the right side of the reservoir container 54. Further, a sealing device 6 is provided on the front side of the reservoir container 54. The sealing device 6 includes a sealing mechanism 30 on which the crystallization plate is mounted and the well opening of the crystallization plate can be sealed using a sealing member.

Here, an example of the crystallization plate used in the sitting drop method is shown in FIG.
As shown in the figure, a plurality of wells 101 are arranged in a lattice pattern on the surface of the crystallization plate 100A. When attention is paid to one well 101, a reservoir well 102 having a large capacity and a drop well 103 having a small capacity are arranged adjacent to each other. In the method of using the crystallization plate 100A, the reservoir solution is dispensed into the reservoir well 102, and the mixed drop of the protein solution and the reservoir solution is dispensed into the drop well 103. Thereafter, the well opening is closed with a sealing member by the sealing mechanism 30 of the sealing device 6 to prevent fluctuation of conditions due to liquid evaporation.

  As described above, in the sitting drop method, the dispensing processing unit 50 is used to prevent the solution in the crystallization plate 100A subjected to the dispensing process from drying, and to attach the seal member to the upper surface of the crystallization plate 100A. Is provided with a seal device 6, and a seal member 30 can be attached to the upper surface of the crystallization plate 100 </ b> A by the seal mechanism 30. The seal mechanism 30 of the seal device 6 includes an actuator including an air cylinder (not shown) for attaching a seal member, and the actuator is appropriately driven according to a signal from the control device 20. Thus, a desired operation required for attaching the seal member is performed. In addition, a single-wafer type sheet is used as the seal member, and the seal mechanism is miniaturized.

An example of a crystallization plate used in the hanging drop method is shown in FIG.
As shown in the figure, a plurality of wells 104 are arranged in a lattice pattern on the surface of the crystallization plate 100B. In the method of using the crystallization plate 100B, the reservoir solution is dispensed into the well 104, a mixed drop of the protein solution and the reservoir solution is prepared on the surface of a separately prepared cover glass, the cover glass is inverted, and the well 104 Cover the opening. Note that a sealant such as grease is applied in advance to the well opening surface 104a, and the opening of the well 104 is sealed only by placing a cover glass.

  As described above, in the hanging drop method, a liquid drop is formed on the surface of the cover glass, and the cover glass is inverted and covered with the opening of the well 104 on the crystallization plate 100B. As shown in FIGS. 3 to 4, a cover glass stock portion 56 on which a plurality of cover glasses can be installed is provided at 50 in front of and on the right side of the sealing device 6 so that these operations can be performed. ing. Furthermore, a cover glass reversing mechanism 57 for reversing the cover glass is provided on the right side of the position where the protein tube 55 is installed. The cover glass reversing mechanism 57 includes a rotary-type air cylinder 57a at the base end, and the air cylinder 57a is appropriately driven according to a signal from the control device 20 to reverse the cover glass. The operation for this is appropriately executed.

  As shown in FIG. 2, a head driving unit 18 provided in the dispensing head driving device 10 is provided in the equipment chamber 3 above the processing chamber 4. The dispensing head driving device 10 is provided with a dispensing head 40 at the tip of the head driving unit 18, and the dispensing head 40 is positioned at a position necessary for the operation in the dispensing processing unit 50. It is possible to move appropriately.

Next, the dispensing head driving device 10 will be described.
Specifically, as shown in FIG. 2A, the dispensing head driving device 10 includes a dispensing head 40 and a head driving unit 18 for moving the dispensing head 40. The head drive unit 18 is driven on the upper surface of the processing chamber upper part 4b so as to be able to move along the bridge portion 16 stretched in the left and right (X) direction in FIG. And a main body (not shown). And this drive part main body 18a has a moving mechanism to the front-back (Y) direction further, Thereby, this dispensing head drive device 10 makes dispensing head 40 each of an X-axis and a Y-axis. It is possible to move to. Further, as shown in FIG. 2B, the dispensing head 40 at the tip of the head driving unit 18 includes a dispensing syringe 42, a liquid level sensor 44, and a cover glass, which are devices necessary for performing a dispensing operation. Adsorption mechanisms 46 are provided so that they can be moved up and down independently in the vertical (Z) direction.

Here, the dispensing syringe 42 can be mounted with a large chip and a small chip disposed in the dispensing processing unit 50, respectively, and can perform dispensing operation of the reservoir solution and the protein solution. Further, the liquid level sensor 44 can detect the level of the reservoir solution in the reservoir container 54 or the protein solution in the protein tube 55, and can set the chip insertion depth when the solution is sucked. ing. Further, the cover glass suction mechanism 46 is used when the hanging drop method is used, and can hold the cover glass by suction. The cover glass suction mechanism 46 is mounted on the head drive unit 18 so that the cover glass is sucked and held and can be moved three-dimensionally. The cover glass suction mechanism 46 and the dispensing head drive device 10 A cover glass adsorption conveyance mechanism is configured.
Thereby, this dispensing head drive device 10 can drive three-dimensionally the device required for performing the dispensing operation provided in the dispensing head 40. The crystallization plates 100A and 100B can be appropriately moved to positions necessary for dispensing into the wells and other positions necessary for work in the dispensing processing unit 50.

Next, the electrical component chamber 5 below the processing chamber 4 will be described.
As shown in FIG. 1, the electrical component room 5 is surrounded by a soundproof material 5 b, and the electrical component room 5 is equipped with a control device 20. The control device 20 is housed in a control box 20b that is formed of a heat insulating material. The control device 20 is sealed by the control box 20b, so that there is no contact with outside air and condensation is prevented. It has become so.

The control device 20 includes control of the sealing mechanism 30 provided in the dispensing head driving device 10 and the sealing device 6, and is configured to be able to control the actuators and sensors provided in the protein crystallization device 1. Further, in the electrical component chamber 5, electrical components necessary for the operation of the protein crystallization apparatus 1, driving of an air cylinder for attaching a sheet in the seal mechanism 30, and a lid 54 a of the reservoir container 54 are provided. Necessary for pneumatic equipment such as compressors necessary for driving each actuator including driving of opening / closing air cylinder 54b and driving of air cylinder 57a when reversing the cover glass, and adsorption of the cover glass A vacuum pump is installed. In other words, all of these electrical components and the like are disposed in the electrical component chamber 5 located below the processing chamber 4, thereby realizing a compact configuration and contributing to a reduction in the installation area of the apparatus. is doing. In addition, since equipment that is often handled as parts outside the apparatus, such as a compressor and a vacuum pump, is also built in the electrical component room 5, an extra space for arranging the compressor and vacuum pump outside the apparatus. Is unnecessary.
Further, the electrical component room 5 is provided with a trash box 9 which is an unnecessary material recovery unit capable of recovering unnecessary materials such as used chips generated by the dispensing operation. The trash box 9 is configured to have a collection port 9a that can be pulled out to the front side of the paper in the figure, and used chips and the like in the trash box 9 can be collected from the collection port 9a.

Next, a sitting drop method performed using the protein crystallization apparatus 1 will be described with reference to FIG. 7 as appropriate.
When implementing the sitting drop method, first, a dispensing material is installed in the dispensing processing unit 50. That is, the open / close door 7 in front of the processing chamber 4 is opened, the handle 8b of the base body 8a is held by hand, and the dispensing processing unit 50 is pulled out of the processing chamber 4. Then, as shown in FIG. 3 described above, with respect to the dispensing processing unit 50, a large chip rack 51 (large chip), a small chip rack 52 (small chip), a reservoir container 54 (reservoir solution), a protein tube 55 (protein) Solution), sheet (seal member), and crystallization plate 100A for sitting drop method are set at predetermined positions, respectively, and then the dispensed processing section 50 is returned into the original processing chamber 4 and the door 7 is opened. Close (step S1A).

Next, a dispensing process is performed. This dispensing process is automatically operated by appropriately driving each part in accordance with a signal from the control device 20.
Specifically, in the dispensing process by the sitting drop method, first, the reservoir solution is dispensed. In dispensing the reservoir solution, the dispensing head 40 is moved above the large tip rack 51, and one large tip among the plurality of the dispensing syringes 42 is pushed into the large tip. Wear by. Next, the dispensing head 40 is moved above the reservoir container 54. The liquid level sensor 44 detects the level of the reservoir solution in one of the plurality of reservoir containers 54. Then, based on the detected liquid level, the large chip is inserted to an appropriate position in the well, and a predetermined amount of the reservoir solution is sucked by the dispensing syringe 42. Next, the dispensing head 40 is moved above the crystallization plate 100A. Then, the reservoir solution is discharged from the large tip into one of the plurality of reservoir wells in the crystallization plate 100A by the dispensing syringe. Then, the dispensing head 40 is moved above the trash box entrance 53. Next, the large tip is detached from the dispensing syringe 42 and discarded from the trash box entrance 53 to the trash box 9 (step S2A).

  Next, a droplet is produced. For the production of the liquid droplets, the dispensing head 40 is moved above the small chip rack 52, and one small chip among a plurality of small chip racks 52 is placed in the dispensing syringe 42. Mount by pushing the tip into a small chip. Next, the dispensing head 40 is moved above the protein tube 55. The liquid level of the protein solution is detected by the liquid level sensor 44. Then, based on the detected liquid level, the small tip is inserted to an appropriate position in the protein tube 55, and a predetermined amount of the protein solution is sucked by the dispensing syringe. Then, the dispensing head 40 is moved above the crystallization plate 100A. Then, a part of a predetermined amount of the reservoir solution discharged to one reservoir well 102 of the crystallization plate 100A is further sucked by the dispensing syringe 42 into the small chip containing the protein solution. As a result, both the protein solution and the reservoir solution are sucked into the small chip. Next, the solution is discharged from the small tip by the dispensing syringe 42 to the drop well 103 adjacent to the reservoir well. Then, the solution in the drop well 103 is repeatedly sucked and discharged to produce a mixed drop (step S3A).

Next, the dispensing head 40 is moved above the trash box entrance 53. The small tip is detached from the dispensing syringe 42 and discarded from the trash box entrance 53 to the trash box 9. Similarly, dispensing of the reservoir solution and preparation of droplets are sequentially performed on the other wells in the same row of the crystallization plate, and a dispensing process for one row of wells is performed (step S4A).
Next, a sealing process is performed. In the sealing process, a sheet is pasted on the upper surface of the crystallization plate 100A by the sealing mechanism unit 30 with respect to the well row subjected to the dispensing process (step S5A).
Then, the above steps S2A to S5A are repeated in the same manner as described above, and the dispensing process and the sealing process for the next well row are sequentially performed (step S6A). When the dispensing process and the sealing process are completed up to the last well row, the crystallization plate 100A subjected to the dispensing seal process is completed.

Next, the hanging drop method performed using the protein crystallization apparatus 1 will be described with reference to FIG. 8 as appropriate.
In carrying out the hanging drop method, first, a dispensing material is installed in the dispensing processing unit 50. That is, the open / close door 7 in front of the processing chamber 4 is opened, the handle 8b of the base body 8a is held by hand, and the dispensing processing unit 50 is pulled out of the processing chamber 4. Then, as shown in FIG. 4 described above, with respect to the dispensing processing unit 50, a large chip rack 51 (large chip), a small chip rack 52 (small chip), a reservoir container 54 (reservoir solution), a protein tube 55 (protein) Solution), the crystallization plate 100B for the hanging drop method, and the cover glass are set at predetermined positions, respectively, and then the dispensed processing section 50 is returned to the original processing chamber 4 and the open / close door 7 is closed (step S1B). ).

Next, a dispensing process is performed. This dispensing process is automatically operated by appropriately driving each part in accordance with a signal from the control device 20.
Specifically, in the dispensing process by the hanging drop method, first, a cover glass is prepared. To prepare the cover glass, first, the dispensing head 40 is moved above the cover glass stock portion 56. Then, one cover glass is adsorbed to the cover glass adsorbing mechanism 46 by the cover glass stock unit 56. Next, the dispensing head 40 is moved above the cover glass reversing mechanism 57. Next, the cover glass is transferred from the cover glass adsorption mechanism 46 to the cover glass reversing mechanism (step S2B).

  Next, the reservoir solution is dispensed. To dispense the reservoir solution, first, the dispensing head 40 is moved above the large tip rack 51, and one large tip among a plurality of large tips rack 51 is attached to the dispensing syringe 42. Next, the dispensing head 40 is moved above the reservoir container 54. Then, the liquid level sensor 44 detects the level of the reservoir solution in one of the plurality of wells. Then, based on the detected liquid level height, the large chip is inserted to an appropriate position in the well, and the reservoir solution is sucked by a predetermined amount by the dispensing syringe 42. Next, the dispensing head 40 is moved above the crystallization plate 100B. Then, the aspirated reservoir solution is discharged from the large tip into one of the plurality of wells in the crystallization plate 100B by the dispensing syringe 42. The dispensing head 40 is moved above the trash box entrance 53. The large tip is detached from the dispensing syringe 42 and discarded from the trash box entrance 53 to the trash box 9 (step S3B).

  Next, a droplet is produced. In producing the droplet, first, the dispensing head 40 is moved above the small chip rack 52, and one small chip among a plurality of small chips rack 52 is mounted on the dispensing syringe 42. Next, the dispensing head 40 is moved above the protein tube 55. Then, the liquid level of the protein solution is detected by the liquid level sensor 44. Then, based on the detected liquid level, the small tip is inserted to an appropriate position in the protein tube 55, and a predetermined amount of the protein solution is sucked by the dispensing syringe. Next, the dispensing head 40 is moved above the crystallization plate 100B. Then, a part of a predetermined amount of the reservoir solution discharged to one well of the crystallization plate 100B is sucked into a small chip containing the protein solution by the dispensing syringe 42. As a result, both the protein solution and the reservoir solution are sucked into the small chip. Then, the dispensing head 40 is moved to above the cover glass reversing mechanism 57. Then, the cover glass is reversed by the cover glass reversing mechanism 57. And a solution is discharged from a small chip | tip with the dispensing syringe 42 with respect to the cover glass surface. The solution on the surface of the cover glass is repeatedly sucked and discharged to produce a mixed drop on the cover glass surface (step S4B).

  Next, it is sealed with a cover glass. For sealing with the cover glass, first, the cover glass is reversed by the cover glass reversing mechanism 57. Thereby, a droplet (mixed drop) will be in the state adhering to the lower surface of the cover glass. Next, the cover glass is transferred from the cover glass reversing mechanism 57 to the cover glass suction mechanism 46 of the dispensing head 40. Then, the dispensing head 40 is moved above the crystallization plate 100B. Then, the cover glass adsorbed by the cover glass adsorption mechanism 46 is placed on the opening surface of one well of the crystallization plate 100B from which the reservoir solution has been discharged. Here, since the sealing agent is applied in advance to the well opening surface, the well is sealed by placing a cover glass. Next, the dispensing head 40 is moved above the trash box entrance 53. Then, the small tip is detached from the dispensing syringe 42 and discarded from the box entrance 53 to the trash box 9 (step S5B).

  Then, steps S2B to S5B are repeated in the same manner as described above. That is, the preparation of the cover glass, the dispensing of the reservoir solution, the preparation of the droplets, and the sealing with the cover glass are sequentially performed on the other wells (step S6B). When these processes are completed up to the last well of the crystallization plate 100B, the crystallization plate 100B subjected to the crystallization dispensing process is completed.

Next, functions and effects of the protein crystallization apparatus 1 will be described.
As described above, according to the protein crystallization apparatus 1, the housing 2 has the processing chamber 4 capable of managing the internal atmosphere in a necessary state, and the processing chamber 4 includes a sealing device. The base 8 (base body 8a) on which the dispensing processing unit 50 including 6 and the like is provided is provided so as to be slidable between the working position located in the processing chamber 4 and the overhanging position protruding outside the processing chamber. Therefore, if the base 8 (base body 8a) is in the overhanging position, the dispensing processing unit 50 on the upper part can be pulled forward. Therefore, as described above, necessary operations such as arrangement of the dispensing material by the sitting drop method and the hanging drop method can be performed outside the processing chamber 4. Therefore, since the work space for manpower in the processing chamber itself can be reduced within a necessary and sufficient range, the apparatus can be miniaturized.

  In other words, even when the processing chamber 4 is configured to be sufficiently small in size, the base 8 can be positioned at the overhanging position, and the dispensing processing unit 50 at the top can be pulled out forward. Therefore, handling and maintenance before and after dispensing work are easy. Therefore, for example, as described above, even when the sitting drop method and the hanging drop method are appropriately changed and used as the vapor diffusion method, it is necessary for each dispensing operation in the dispensing processing unit 50. Necessary operations such as replacement of the dispensing material can be easily performed, and the same applies to the sealing device 6 on which the corresponding crystallization plates 100A to 100B are placed on the dispensing processing unit 50. In addition, it is possible to easily replace the crystallization plates 100A to 100B corresponding to each dispensing operation. Therefore, according to the protein crystallization apparatus 1, when a plurality of vapor diffusion methods are appropriately changed or when it is necessary to produce a small number of crystallization plates having different specifications, a so-called multi-product small lot requirement is required. However, it is easy to use.

Further, according to the protein crystallization apparatus 1, the casing 2 has a dispensing head driving device 10, a base 8 provided with a dispensing processing unit 50 on the upper surface, and control in the electrical component chamber 5 below the base. Since the device 20 and the trash can 9 are installed in this order from the top of the housing 2, a so-called compact configuration can be realized by this arrangement, which contributes to further downsizing of the device.
Furthermore, according to the protein crystallization apparatus 1, the processing chamber 4 has the open / close door 7 formed of a material that can visually recognize the processing chamber. The state can be easily confirmed. Further, since the base 8 can be positioned at the extended position by projecting the opening / closing door 7 from the opened portion to the outside of the processing chamber 4, the apparatus can be downsized. It is more suitable as a configuration that can improve the usability.

Moreover, according to this protein crystallization apparatus 1, since the whole protein crystallization apparatus 1 is installed in a cool box and dispensed, the protein crystallization apparatus 1 itself has, for example, a Peltier element or the like. The protein crystallization apparatus 1 can be configured without providing a cooling mechanism. Therefore, it can be reduced in size with a minimum configuration.
And since the entire protein crystallization apparatus 1 is placed in a cool box and dispensed, all of the solution and the container, the tip in contact with the solution, the syringe, the dispensing plate, the glass plate, etc. can be cooled uniformly. Thus, denaturation of the protein solution can be prevented and the quality of crystallization can be improved. In the case of partial cooling by a cooling mechanism having the above Peltier element, etc., temperature unevenness occurs, so that the denaturation of the protein solution is more suitably prevented and the quality of crystallization is improved. The configuration of the form is preferred. And since the preset temperature of the said cool box is set to 4 degreeC and dispensing operation is performed at the preset temperature, it is more suitable when improving the quality of crystallization.

Moreover, according to this protein crystallization apparatus 1, since the periphery of the control device 20 is hermetically sealed with the heat insulating material 20b, the electrical component in the control device 20 is prevented from dew condensation so as to prevent the electrical component from condensing. can do.
Furthermore, the electrical component chamber 5 is provided with pneumatic devices such as a compressor necessary for driving each actuator, and a vacuum pump necessary for adsorption of the cover glass. Since the surroundings are surrounded by the soundproofing material 5b, noise generated from the installed devices can be reduced. In the example of this embodiment, the noise at the front of the protein crystallization apparatus 1 is suppressed to 50 dB or less.

Furthermore, according to the protein crystallization apparatus 1, since the electrical component chamber 5 and the processing chamber 4 are separated from each other by the heat insulating material 5a, the entire protein crystallization apparatus 1 is provided as in the example of this embodiment. Is placed in the cool box and dispensed, the heat in the electrical component room 5 is prevented from being transferred to the process chamber 4, and the temperature in the process chamber 4 is kept at the set temperature of the cool box. It is suitable for doing so.
As described above, according to this protein crystallization apparatus 1, it is possible to provide a protein crystallization apparatus that is small and easy to use.

The protein crystallization apparatus according to the present invention is not limited to the above embodiment, and various modifications can be made without departing from the spirit of the present invention.
For example, in the above-described embodiment, the entire protein crystallization apparatus 1 is installed in the cool box and dispensed, and the temperature in the processing chamber 4 is maintained at the set temperature of the cool box. However, the present invention is not limited to this. For example, the entire protein crystallizing apparatus 1 is not installed in the cold storage, and only the temperature in the processing chamber 4 is set to a desired set temperature (for example, 4 ° C.). Good. However, in order to perform crystallization with better quality, it is preferable that the entire protein crystallization apparatus 1 is installed in a cool box and the dispensing operation is performed while the whole is uniformly cooled.

  Further, for example, in the above-described embodiment, the open / close door 7 has been described with an example in which the open / close doors 7a and 7b are formed of transparent tempered glass so that the inside of the processing chamber can be visually recognized. However, the present invention is not limited thereto. For example, a door made of steel plate may be used. However, in order to make it possible to easily check the state of the processing chamber from the outside through the opening and closing door, it is preferable to form the opening and closing door with a material that can visually recognize the processing chamber. In the above-described embodiment, the open / close door 7 has been described as an example of a double door opening. However, the present invention is not limited thereto, and may be, for example, a sliding door.

  Further, for example, in the above embodiment, the slidable base 8 (base body 8a) has been described as an example of sliding in the direction opposite to the position where the open / close door 7 is opened, but the present invention is not limited to this. An opening other than the opening / closing door 7 may be provided separately and slidable with respect to the dedicated opening. However, in order to reduce the size of the apparatus in a compact configuration, it is preferable that the base can be positioned at the extended position by protruding outside the processing chamber from a portion where the opening / closing door is opened.

It is the external view which looked at one Embodiment of the protein crystallization apparatus which concerns on this invention from the front. It is a figure of the state which removed the exterior cover containing the opening-and-closing door in FIG. FIG. 5 is a layout view of a dispensing processing unit when a sitting drop method is used in the protein crystallization apparatus of the present embodiment. FIG. 3 is a layout view of a dispensing processing unit when a hanging drop method is used in the protein crystallization apparatus of the present embodiment. It is a figure which shows an example of the crystallization plate used for the sitting drop method. It is a figure which shows an example of the crystallization plate used for the hanging drop method. It is a figure explaining the operation | work at the time of using the sitting drop method with the protein crystallization apparatus of this embodiment. It is a figure explaining the operation | work at the time of using the hanging drop method with the protein crystallization apparatus of this embodiment.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Protein crystallizer 2 Case 3 Equipment room 4 Processing room 5 Electrical component room 6 Sealing device 7 Opening / closing door 8 Base 9 Trash box (unnecessary substance collection part)
DESCRIPTION OF SYMBOLS 10 Dispensing head drive device 16 Bridging part 18 Head drive part 20 Control apparatus 22 Base drawer mechanism 30 Sealing mechanism part 40 Dispensing head 42 Dispensing syringe 44 Liquid level sensor 46 Cover glass adsorption mechanism 50 Dispensing process part 51 Large chip Rack 52 Small chip rack 53 Trash entrance 54 Reservoir container 55 Protein tube 56 Cover glass stock part 57 Cover glass reversing mechanism 100A, 100B Crystallization plate 101 Well 102 Reservoir well 103 Drop well 104 Well

Claims (7)

  Sliding movement between a housing having a processing chamber for performing a desired dispensing operation on the crystallization plate, a working position provided in the housing and positioned in the processing chamber, and an overhanging position protruding outside the processing chamber Possible pedestal, the crystallization plate provided on the pedestal, a sealing device capable of sealing the well opening of the crystallization plate using a seal member, and the dispensing required for the dispensing operation Dispensing processing section in which materials are respectively disposed, and a dispensing head provided above the dispensing processing section in the processing chamber for performing necessary processing such as dispensing on the well of the crystallization plate A dispensing head driving device capable of moving the dispensing head to a position necessary for processing the well of the crystallization plate, and the dispensing head driving device provided below the base. Shi A control device for controlling Le device, respectively, and protein crystallization apparatus characterized in that it comprises a electrical component compartment, a with the dispensing unwanted matter recovery unit which is capable of recovering the unnecessary ones caused by the work.   The processing chamber has an opening / closing door formed of a material that allows the processing chamber to be visually recognized, and the base can be positioned at the extended position by protruding outside the processing chamber from a portion where the opening / closing door is opened. The protein crystallization apparatus according to claim 1, wherein:   The protein crystallization apparatus according to claim 1 or 2, wherein the entire protein crystallization apparatus is placed in a cool box and dispensed.   The protein crystallization apparatus according to claim 3, wherein a set temperature of the cool box is set to 4 ° C, and a dispensing operation is performed at the set temperature.   The protein crystallization apparatus according to claim 1, wherein the controller is hermetically sealed with a heat insulating material.   The protein crystallization apparatus according to claim 1, wherein the electrical component chamber is surrounded by a soundproofing material.   The protein crystallization apparatus according to claim 1, wherein the electrical component chamber and the processing chamber are partitioned from each other by a heat insulating material.

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