JP3572322B2 - Equipment for making hanging drops of protein crystallization - Google Patents

Description

[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to an apparatus for producing a protein crystallization hanging drop used in structural genomics.
[0002]
[Prior art]
2. Description of the Related Art In recent years, structural biology, in particular, structural genomics for clarifying the three-dimensional structure of a protein and physicochemically elucidating all functions of a living body has been actively pursued.
[0003]
In order to clarify the three-dimensional structure of a protein, first, a test for obtaining a protein crystal by crystallizing the protein is performed, and then the three-dimensional structure is measured by X-ray diffraction of the obtained protein crystal. .
[0004]
Therefore, in order to clarify the three-dimensional structure of a protein, it is necessary to first produce a crystal of the protein, and as a method of crystallizing the protein, a hanging drop vapor diffusion method in which good quality crystals are easily obtained is known. .
[0005]
The hanging drop vapor diffusion method hangs a mixed drop, in which a protein solution and a crystallization solution are mixed, on the upper wall in a closed container filled with the crystallization solution, and mixes the mixed drops separated in the gas phase with excess crystals. By utilizing the equilibration of the crystallization agent concentration with the agent solution, crystallization is realized by slowly creating a supersaturated state of the protein in the mixed drop.
[0006]
The protein crystallization method is a method in which crystallization is attempted using a representative crystallization agent that has succeeded in crystallizing various proteins known so far. For proteins, it is necessary to repeat the operation of sequentially changing the type of the crystallization agent, further changing the concentration of each crystallization agent, and searching for the crystallization conditions, and an enormous number of tests are required.
[0007]
[Problems to be solved by the invention]
However, the protein crystallization test as described above is complicated and requires a great deal of trouble. Therefore, if such complicated protein crystallization tests were repeatedly performed manually for the huge number required from the search for the initial conditions to the establishment of the optimal conditions, it would require a huge amount of personnel and time. It cannot meet the demands of the development of structural genomics.
[0008]
Further, since the protein solution is expensive, it is necessary to minimize the amount of protein used in one crystallization drop in many crystallization tests. Therefore, since it is necessary to handle the protein solution and the crystallization agent in a very small amount and with high precision, the handling operation is technically difficult, and there is a problem that the working efficiency is further reduced.
[0009]
Therefore, it is very important to save labor and increase the efficiency of the protein crystallization test process in order to promote structural genomics.
[0010]
The present invention has been made in view of the above problems, and has as its object to provide an apparatus for preparing a protein crystallization hanging drop, which enables a protein crystallization test to be performed efficiently with low labor.
[0011]
[Means for Solving the Problems]
The present invention has a plurality of solution pockets, and a drop making pallet that can seal the upper opening of each solution pocket with an upper glass,
A glass reversal support device that supports the upper glass horizontally and is capable of reversing the upper and lower surfaces of the upper glass,
An end effector fixing plate that can move back and forth, left and right, and up and down at the top of the fixed drop making pallet;
A medium amount solution which is attached to the end effector fixing plate so as to be able to move up and down, and a predetermined amount of the crystallizing agent solution of the crystallizing agent solution container separately provided by a replaceable medium amount suction tip and supplied to an arbitrary solution pocket. A dispensing device,
A small amount of protein solution is aspirated from the protein solution container attached separately to the end effector fixing plate so as to be movable up and down by a replaceable micro suction tip and supplied onto the upper glass on the glass reversing support device. Forming a drop, and a small amount solution dispensing device that forms a mixed drop by supplying a small amount of the crystallization agent solution by another small amount suction tip and mixing it with the protein drop,
A grease applying device attached to the end effector fixing plate so as to be able to move up and down, and applying grease to an upper opening of the solution pocket,
A glass attachment / detachment device which is attached to the end effector fixing plate so as to be able to move up and down, grasps the upper glass of the glass inversion support device, and mounts the glass at the upper opening of the solution pocket;
The present invention relates to an apparatus for producing a hanging drop for protein crystallization, comprising:
[0012]
In the above means, a tip detector for detecting at least the tip position of the micro suction suction tip of the micro suction tip and the medium suction tip may be provided, and the liquid level of the protein solution container and the crystallization agent A liquid level detecting device for detecting at least the liquid level of the protein solution container among the liquid surfaces of the solution container may be provided.
[0013]
A container body having a solution pocket having an annular liquid chamber on the outer periphery of the center tube, an outer wall of the annular liquid chamber extending above the center tube, and having an upper opening formed therein; A lower glass placed on the upper end of each central tube of the container body to close the central tube, and an upper glass placed on the upper opening of the solution pocket to close each solution pocket, wherein the grease applying device is Grease may be applied to the upper end opening of the tube, and the glass attaching / detaching device may place the lower glass on the upper end opening of the central tube.
[0014]
According to the present invention, the following operation is performed.
[0015]
Operation of supplying the crystallization solution to the solution pocket of the drop making pallet, operation of supplying the protein solution and the crystallization solution to the upper glass to form a mixed drop, the upper end of the center tube and the upper opening of the solution pocket The operation of applying grease to, the operation of inverting the upper glass, the operation of placing the lower glass and the upper glass on the upper end of the central tube and the upper opening of the solution pocket, and sealing the solution pocket can all be automated, Therefore, the protein crystallization test can be performed efficiently with low labor.
[0016]
In addition, by providing a liquid level detection device and a tip detector and supplying a small amount of a protein solution and a crystallization agent solution by a small solution dispensing device, it is possible to accurately prepare a small mixed drop. Yes, thus allowing for testing with small protein solutions.
[0017]
BEST MODE FOR CARRYING OUT THE INVENTION
Hereinafter, preferred embodiments of the present invention will be described with reference to the drawings.
[0018]
The present invention realizes labor saving and high efficiency by automating the operation of preparing a protein crystallizing hanging drop when crystallizing a protein using the hanging drop vapor diffusion method.
[0019]
FIG. 1 is a plan view showing an example of a drop making pallet used in the protein crystallization hanging drop making apparatus of the present invention, and FIG. 2 is a view taken along the line II-II in FIG.
[0020]
The drop making pallet 1 shown in FIGS. 1 and 2 has a plurality of (for example, about 50) solution pockets 4 arranged in a laterally and horizontally aligned manner inside an outer wall 3 having an overhanging portion 2 formed on the upper periphery. Is formed of a synthetic resin material.
[0021]
As shown in FIG. 3, each solution pocket 4 of the container body 1 </ b> A is provided with a center tube 5 formed vertically and having an upper end extending to an upper and lower middle position of the outer wall 3 at the center. A peripheral wall 7 forming an annular liquid chamber 6 is integrally provided on the outer periphery. The peripheral wall 7 extends above the upper end 5a of the central tube 5 to form an upper opening 7a.
[0022]
Further, the drop making pallet 1 is placed on the upper end 5a of the center tube 5 of each solution pocket 4 so as to close the center tube 5, and is placed on the upper opening 7a of the solution pocket 4. And an upper glass 9 for closing each solution pocket 4. The central tube 5 provided in the solution pocket 4 is illuminated from below through the central tube 5 and the lower glass 8 so that crystals can be observed to be formed on a hanging drop formed on the lower surface of the upper glass 9. ing.
[0023]
The drop making pallet 1 is placed on a pallet support 10 so as to be held substantially horizontally.
[0024]
As shown in FIGS. 4 and 5, a plurality of crystals containing a plurality of types of crystallization agent solutions are placed on a work table 11 on the side of the drop making pallet 1 supported on the pallet support 10. The agent solution container 12 is arranged. As the crystallization agent, a known representative crystallization agent which has succeeded in crystallization of a protein can be used.
[0025]
Further, a plurality of protein solution containers 13 containing a plurality of types of protein solutions to be tested are arranged on the side of the crystallization agent solution device 12.
[0026]
A plurality of upper glasses 9 are laminated and stocked on the side of the crystallization agent solution container 12, and an upper glass dispenser 14 that dispenses one by one, and a plurality of lower glasses 8 are laminated and stocked. And a lower glass dispensing machine 15 for dispensing one by one is installed.
[0027]
A glass reversing support device 17 is provided on a side portion of the upper glass dispenser 14, and the glass reversing support device 17 receives the upper glasses 9 discharged from the upper glass dispenser 14 one by one. The upper and lower surfaces of the upper glass 9 are inverted by rotating horizontally by 180 ° about a horizontal rotation shaft 16.
[0028]
Further, a lower glass support device 18 is provided on a side portion of the lower glass dispenser 15, and the lower glass support device 18 receives the lower glasses 8 discharged from the lower glass dispenser 15 one by one and horizontally supports them. I have to.
[0029]
A fixed frame 20 supported by support legs 19 is provided at an upper portion of the work table 11. The fixed frame 20 includes a left-right movable table 21 that moves in the left-right direction X along the fixed frame 20. A front-rear moving table 22 that moves in the front-rear direction Y along the left-right moving table 21 is provided. The front-rear moving table 22 is provided with a vertical moving table 45 that moves in the vertical direction Z. The end effector fixing plate 23 is fixed to 45.
[0030]
Thereby, the end effector fixing plate 23 can move back and forth, left and right, and up and down the upper part of the drop making pallet 1, the crystallization agent solution device 12, the protein solution device 13, the glass inversion support device 17 and the lower glass support device 18. It has become.
[0031]
FIG. 6 is a front view showing details of the end effector fixing plate 23, and FIG. 7 is a side view of FIG. 6 as seen from the VII-VII direction.
[0032]
The end effector fixing plate 23 includes a liquid level detecting device 24 for detecting the liquid level of the crystallization agent solution device 12 and the liquid level of the protein solution device 13, and a medium solution dispensing device 25. , A small amount solution dispensing device 26 and a glass attaching / detaching device 27 are arranged in a straight line.
[0033]
The liquid level detecting device 24 is provided at the lower raising / lowering end of a lifting / lowering device 28 such as an air cylinder provided vertically on the end effector fixing plate 23, the liquid level of the crystallization agent solution device 12 or the protein solution device 13. An ultrasonic distance meter 29 for detecting the liquid level of the liquid level is provided. Further, as the liquid level detecting device 24, a device which detects the liquid level using light or the like other than the ultrasonic distance meter 29 may be used. The liquid level detecting device 24 in FIG. 6 has a configuration in which the ultrasonic distance meter 29 is surrounded by a cylinder 29a. In this way, when the ultrasonic range finder 29 is configured to be surrounded by the cylinder 29a, the spread of the ultrasonic waves can be suppressed, and the liquid level can be detected with high accuracy.
[0034]
The medium-volume solution dispensing device 25 is configured by attaching a supply / discharge device 31 connected to a known syringe pump (not shown) to a lower lifting end of a lifting device 30 such as an air cylinder provided vertically on the end effector fixing plate 23. In addition, a medium amount suction tip 32 is exchangeably mounted below the supply / discharge device 31.
[0035]
The small amount solution dispensing device 26 has a supply / discharge device 34 connected to a well-known syringe pump (not shown) attached to a lower lifting end of a lifting device 33 such as an air cylinder provided vertically on the end effector fixing plate 23. At the lower part of the supply / discharge device 34, a small amount of suction tip 35 is exchangeably mounted.
[0036]
The glass attachment / detachment device 27 has a suction pipe (not shown) attached to a lower elevating end of a lifting / lowering device 36 such as an air cylinder provided vertically on the end effector fixing plate 23, and a suction port provided at a lower end of the suction pipe. 37, the upper glass 9 on the glass reversing support device 17 and the lower glass 8 on the lower glass support device 18 are sucked and gripped, and the upper opening 7a of the solution pocket 4 in FIG. 5 is placed on the upper end 5a to perform closing.
[0037]
A grease applying device 38 is provided at a position on the end effector fixing plate 23 before the glass attaching / detaching device 27. As shown in FIGS. 5 to 7, the grease applying device 38 includes an elevating device 39 that can move vertically along the end effector fixing plate 23. The grease connected to a dispenser (not shown) is connected to the elevating device 39. A syringe 40 is mounted, and a grease needle 40a is mounted below the grease syringe 40. The grease needle 40 a applies grease to the upper opening 7 a of the solution pocket 4 and the upper end 5 a of the center tube 5 in the drop making pallet 1.
[0038]
At a required position of the worktable 11, a tip detector 41 such as a magnescale for detecting the tip position (lower end position) of the medium amount suction tip 32 and the small amount suction tip 35 is provided. ing.
[0039]
The operation of the above embodiment will be described below.
[0040]
In making the protein crystallizing hanging drop, the drop making pallet 1 shown in FIG. 1 and FIG. 2 is placed on the pallet support 10 in FIG. To support.
[0041]
Subsequently, a predetermined amount of the crystallizing agent solution of the crystallizing agent solution dispenser 12 at a predetermined position is suctioned by the medium amount suction tip 32 of the medium amount solution dispensing device 25 in FIGS. 1 to the solution pocket 4 at a predetermined position.
[0042]
At this time, in order to supply a specified amount of the crystallization agent solution, it is necessary to measure in advance the amount (liquid level) of the liquid contained in the predetermined crystallization agent solution device 12. That is, when the medium amount suction tip 32 sucks the crystallization agent solution, if the insertion amount of the medium amount suction tip 32 with respect to the liquid level changes, the actual suction amount changes even if the constant suction is performed. I will.
[0043]
For this purpose, first, an operation of moving the left and right movable table 21 in the left and right direction X along the fixed gantry 20, an operation of moving the end effector fixing plate 23 in the front and rear direction Y along the left and right movable table 21, and By moving the end effector fixing plate 23 in the vertical direction Z along the table 21, the end effector fixing plate 23 is moved back and forth, left and right, and up and down, and the liquid level detecting device 24 is moved to the predetermined crystallization agent solution container. After moving to just above 12, the ultrasonic range finder 29 is lowered by the elevating device 28 to measure the liquid level L1 (FIG. 8). At this time, since the ultrasonic distance meter 29 is surrounded by the cylinder 29a, the spread of the ultrasonic wave is suppressed, and the liquid level is detected with high accuracy.
[0044]
In addition, the medium amount suction tip 32, which is exchangeably attached to the supply / discharge device 31 of the medium amount solution dispensing device 25, has a length dimension error due to manufacturing or the like. In order to supply the crystallization agent solution, the lower end position of the medium amount suction tip 32 needs to be measured in advance. For this purpose, the medium amount suction tip 32 is moved by the end effector fixing plate 23 so as to be directly above the tip detector 41, and then the medium amount suction tip 32 is lowered by the elevating device 30, and the medium amount The lower end position of the suction tip 32 is measured, and this position is set as the origin (zero point).
[0045]
After performing the above-described measurement, the medium amount suction tip 32 is moved to a position immediately above the predetermined crystallization agent solution device 12, and the medium amount suction tip 32 is lowered by the elevating device 30. At this time, the liquid level height L1 of the crystallization agent solution device 12 is measured as described above, and the tip position of the medium amount suction tip 32 is measured. As), the tip of the medium amount suction tip 32 is inserted into the solution by a predetermined length S1 from the liquid level L1, and a predetermined amount of the crystallization agent solution is suctioned in this state. By accurately setting the length S1 at which the tip of the medium amount suction tip 32 is inserted from the liquid level L1, the suction amount can be controlled with high accuracy and high reproducibility, and the insertion length can be controlled. By setting S1 to be small, the amount of the crystallization agent solution adhering to the outer surface of the tip of the medium amount suction tip 32 is also reduced, so that more accurate suction is possible.
[0046]
Subsequently, the medium suction tip 32 in a state where the crystallization agent solution is sucked is raised, and the end effector fixing plate 23 is moved so that the medium suction tip 32 is positioned above the solution pocket 4 at a predetermined position on the drop forming pallet 1. Then, the medium suction tip 32 is lowered to supply the crystallization agent solution to the annular liquid chamber 6 of the solution pocket 4 shown in FIG.
[0047]
In the above, when the accuracy of the amount of the crystallization agent solution supplied to the solution pocket 4 is not required, the operation of measuring the lower end position of the medium amount suction tip 32 may be omitted.
[0048]
Next, on the upper surface of the upper glass 9 which has been dispensed from the upper glass dispenser 14 onto the glass reversing support device 17, the trace suction tip 35 of the trace solution dispensing device 26 shown in FIGS. 4 and 6 is used. A small amount of the protein solution and a small amount of the same crystallization agent solution device 12 as described above are supplied to perform an operation of forming a mixed drop.
[0049]
At this time, in order to accurately aspirate a small amount of protein solution for forming a hanging drop from the protein solution device 13 and supply it onto the upper glass 9, the amount (liquid amount) of the protein solution stored in the protein solution device 13 Surface height) must be measured in advance. That is, when the amount of insertion of the minute amount suction tip 35 with respect to the liquid surface changes when the protein solution is sucked by the minute amount suction tip 35, the actual amount of suction changes even if the constant suction is performed.
[0050]
For this purpose, the liquid level detecting device 24 is moved directly above the predetermined protein solution container 13 by the end effector fixing plate 23, and then the ultrasonic range finder 29 is lowered by the elevating device 28 so that the liquid level height L2 ( FIG. 9) is measured.
[0051]
Further, the micro suction tip 35, which is exchangeably attached to the supply / discharge device 34 of the micro solution dispenser 26, has a length dimension error due to manufacturing or the like. In order to supply a protein solution with high accuracy, it is necessary to measure the lower end position of the micro suction tip 35 in advance. To this end, after moving the micro suction tip 35 by the end effector fixing plate 23 so as to be directly above the tip detector 41, the micro suction suction tip 35 is lowered by the elevating device 33, and the micro suction tip 35 is moved. Is measured, and this position is set as the origin (zero point).
[0052]
After performing the above measurement, the micro suction tip 35 is moved to a position immediately above the predetermined protein solution device 13, and the elevating device 33 lowers the micro suction tip 35. At this time, since the liquid level height L2 of the protein solution device 13 in FIG. 9 is measured and the tip position of the micro suction tip 35 is measured, the tip position is set as the origin (zero point). The tip of the suction tip 35 is inserted into the solution by a predetermined length S2 from the liquid level L2, and a predetermined amount of the protein solution is suctioned in this state.
[0053]
In this way, by accurately setting the length S2 at which the tip of the micro suction tip 35 is inserted from the liquid level L2, it becomes possible to control the suction amount with high accuracy and high reproducibility. Further, by setting the insertion length S2 to be small, the amount of the protein solution adhering to the outer surface of the distal end of the micro suction tip 35 is reduced, so that more accurate suction can be performed.
[0054]
After raising the micro suction tip 35 and further moving the micro suction tip 35 to the upper portion of the upper glass 9 by the end effector fixing plate 23, the sucked protein solution is placed on the upper glass 9 as shown in FIG. By supplying, first, a protein drop 42 is formed.
[0055]
Subsequently, the liquid level height of the crystallization agent solution device 12 is measured by the ultrasonic distance meter 29 of the liquid level detecting device 24, and the tip position of the newly replaced trace suction tip 35 is detected by the tip detector 41. After the measurement, the crystallization agent solution of the crystallization agent solution device 12 is supplied by the micro suction tip 35 to the length S2 at which the tip of the micro suction tip 35 is inserted from the liquid level L2 as in FIG. Is suctioned in a state where is set to be small, and thereafter, the mixture is supplied to the protein drop 42 on the upper glass 9 shown in FIG.
[0056]
In the above, the crystallization agent solution may be a single component solution or a solution in which a plurality of components are mixed. At this time, the crystallization agent solution forming the mixed drop 43 is supplied to the corresponding solution pocket 4. Must match the crystallization agent solution used.
[0057]
On the other hand, a plurality of crystallization agent solutions may be supplied to the solution pockets 4 of the drop making pallet 1 and mixed. In this case, it is necessary to supply a small amount of the mixed crystallization agent solution in the solution pocket 4 to the protein drop 42 by suction. Therefore, in this case, it is necessary to measure the liquid level of the crystallization agent solution in the solution pocket 4 in order to accurately aspirate the crystallization agent solution in the solution pocket 4. As described above, the liquid level of the crystallization agent solution in the solution pocket 4 can also be favorably measured by the ultrasonic distance meter 29.
[0058]
When aspirating the crystallizing agent solution in the solution pocket 4, the tip for a minute amount of suction tip 35 is replaced with a new one, and the tip position of the tip for the minute amount of suction tip 35 is measured by the tip detector 41. The liquid level 29 in the solution pocket 4 is measured by moving the ultrasonic wave meter 29 right above the predetermined mixing drop 43 and the ultrasonic distance meter 29 (FIG. 9). Subsequently, the crystallization agent solution in the solution pocket 4 is sucked in a state in which the tip S of the microscopic suction tip 35 is set so that the length S2 inserted from the liquid level L2 is reduced as in FIG. Thereafter, the mixture is supplied to the protein drop 42 on the upper glass 9 shown in FIG.
[0059]
As described above, the liquid level detecting device 24 and the tip detector 41 are provided, and the protein solution and the crystallization agent solution are configured to be supplied by the trace solution dispensing device 26 in a very small amount. Can be accurately prepared. Therefore, a test with a small amount of a protein solution becomes possible.
[0060]
Next, after moving the grease applying device 38 onto the upper end 5a of the central tube 5 in the solution pocket 4 to which the crystallization agent solution has been supplied, the lifting device 39 moves the grease needle 40a to the upper end 5a of the central tube 5. The grease 44 (FIG. 10) is applied to the upper end 5a by moving the grease needle 40a along the annular shape of the center tube 5 by moving the end effector fixing plate 23 back and forth, right and left, and up and down. Further, grease 44 (FIG. 10) is applied to upper opening 7a of solution pocket 4 by grease applying device 38 in the same manner as described above.
[0061]
Subsequently, the glass attachment / detachment device 27 is moved onto the lower glass support device 18 by the movement of the end effector fixing plate 23, and the one lower glass 8 paid out from the lower glass dispenser 15 to the lower glass support device 18 is attached to the glass attachment / detachment device. Then, the glass attachment / detachment device 27 is moved to the upper portion of the solution pocket 4 to which the crystallization solution is supplied, and the lower glass 8 is moved to the upper end 5a of the central tube 5 by the elevating device 36. The grease 44 is pressed and placed on the applied grease 44.
[0062]
Further, by rotating the upper glass dispensing machine 14 supporting the upper glass 9 (indicated by phantom lines in FIG. 10) on which the mixing drops 43 are formed by 180 ° about the rotation shaft 16, the mixing drops 43 are lowered. Flip it to the side. In this state, the glass attachment / detachment device 27 is moved to the upper part of the upper glass 9 and the upper glass 9 on the glass reversing support device 17 is sucked and gripped by the suction port 37 of the glass attachment / detachment device 27. The upper glass 9 is moved to the upper part of the solution pocket 4 and pressed by the elevating device 36 onto the grease 44 applied to the upper opening 7a of the solution pocket 4 and placed.
[0063]
This forms a sealed solution pocket 4 as shown in FIG. The above operation is performed for each of the solution pockets 4 of the drop making pallet 1 in FIG.
[0064]
Between the above-mentioned crystallization agent solution in the annular liquid chamber 6 in the sealed solution pocket 4 and the mixed drop 43 (hanging drop) formed of the same crystallization agent solution and protein solution formed in the upper glass 9. Then, a vapor diffusion action is performed, and a moderate supersaturation state of the protein is created in the hanging drop to realize crystallization. At this time, by illuminating from the lower part through the lower glass 8 from the central tube 5 provided in the solution pocket 4, the crystal formation in the hanging drop is observed.
[0065]
According to the above-described apparatus for producing a protein crystallization hanging drop, the operation of supplying the crystallization agent solution to the solution pocket 4 of the drop preparation pallet 1 and the step of supplying the protein solution and the crystallization agent solution onto the upper glass 9 are performed. An operation for forming the mixing drop 43, an operation for applying grease 44 to the upper end 5a of the central tube 5 and the upper opening 7a of the solution pocket 4, an operation for inverting the upper glass 9, the lower glass 8 and the upper glass 9 for the upper end of the central tube 5. 5a and the operation of placing the solution pocket 4 on the upper opening 7a of the solution pocket 4 to seal the solution pocket 4 can be fully automated, and therefore, the protein crystallization test can be performed efficiently with less labor.
[0066]
In addition, the liquid level detecting device 24 and the tip detector 41 are provided, and a small amount of the protein solution and the crystallization agent solution are supplied by the small amount solution dispensing device 26 so that the minute mixing drop 43 can be accurately detected. It can be prepared with good reproducibility, and thus enables testing with a small amount of protein solution.
[0067]
It should be noted that the present invention is not limited only to the above-described embodiment, and various changes can be made without departing from the gist of the present invention.
[0068]
【The invention's effect】
According to the present invention, an operation of supplying a crystallization agent solution to a solution pocket of a drop making pallet, an operation of supplying a protein solution and a crystallization agent solution on an upper glass to form a mixed drop, and an upper end of a central tube And applying the grease to the upper opening of the solution pocket, inverting the upper glass, placing the lower glass and the upper glass on the upper end of the central tube and the upper opening of the solution pocket, and sealing the solution pocket. It can be automated, and thus has an effect that the protein crystallization test can be performed efficiently with low labor.
[0069]
In addition, a liquid level detector and a tip detector are provided, and a small amount of a protein solution and a crystallization agent solution are supplied by a small amount solution dispenser, so that a small amount of mixed drops can be accurately and reproducibly prepared. Therefore, there is an effect that a test with a small amount of a protein solution can be performed.
[Brief description of the drawings]
FIG. 1 is a plan view showing an example of a drop making palette in a protein crystallizing hanging drop making apparatus of the present invention.
FIG. 2 is a view in the direction of arrows II-II in FIG.
FIG. 3 is a cross-sectional view illustrating an example of a solution pocket.
FIG. 4 is a front view of the apparatus for producing a protein crystallization hanging drop of the present invention.
FIG. 5 is a plan view of FIG. 4;
FIG. 6 is a front view showing details of an end effector fixing plate.
FIG. 7 is a side view of FIG. 6 as viewed from the direction of VII-VII.
FIG. 8 is a cross-sectional view of a crystallization agent solution container.
FIG. 9 is a sectional view of a protein solution container.
FIG. 10 is a sectional view showing a state in which the solution pocket is sealed and a hanging drop is formed.
[Explanation of symbols]
1 Drop creation palette
1A Container body
3 outer wall
4 Solution pocket
5 Central tube
5a Top
6 annular liquid chamber
7 Perimeter wall
7a Upper opening
8 Lower glass
9 Upper glass
12 Crystallizing agent solution vessel
13 Protein solution container
17 Glass reversal support device
23 End effector fixing plate
24 Liquid level detector
25 Medium volume solution dispenser
26 Micro solution dispenser
27 Glass attachment / detachment device
32 Medium volume suction tip
35 Micro suction tip
38 Grease application device
41 Advanced detector
42 protein drop
43 Mixed Drop
44 Grease

Claims (5)

A drop making pallet having a plurality of solution pockets, and capable of closing an upper opening of each solution pocket with an upper glass,
A glass reversal support device that supports the upper glass horizontally and is capable of reversing the upper and lower surfaces of the upper glass,
An end effector fixing plate that can move back and forth, left and right, and up and down at the top of the fixed drop making pallet;
A medium amount solution which is attached to the end effector fixing plate so as to be able to move up and down, and a predetermined amount of the crystallizing agent solution of the crystallizing agent solution container separately provided by a replaceable medium amount suction tip and supplied to an arbitrary solution pocket. A dispensing device,
A small amount of protein solution is aspirated from the protein solution container attached separately to the end effector fixing plate so as to be movable up and down by a replaceable micro suction tip and supplied onto the upper glass on the glass reversing support device. Forming a drop, and a small amount solution dispensing device that forms a mixed drop by supplying a small amount of the crystallization agent solution by another small amount suction tip and mixing it with the protein drop,
A grease applying device attached to the end effector fixing plate so as to be able to move up and down, and applying grease to an upper opening of the solution pocket,
A glass attachment / detachment device which is attached to the end effector fixing plate so as to be able to move up and down, grasps the upper glass of the glass inversion support device, and mounts the glass at the upper opening of the solution pocket;
An apparatus for producing a protein crystallizing hanging drop, comprising: 2. The apparatus for producing a protein crystallization hanging drop according to claim 1, further comprising a tip detector for detecting a tip position of at least a tip of the trace suction tip among the trace suction tip and the medium suction tip. . The protein according to claim 1, further comprising a liquid level detection device that detects at least a liquid level of the protein solution container among a liquid surface of the protein solution container and a liquid surface of the crystallization agent solution container. Equipment for making crystallization hanging drops. 4. The apparatus according to claim 3, wherein the liquid level detecting device has a configuration in which the ultrasonic distance meter is surrounded by a cylinder. A container body having a liquid pocket on an outer periphery of a center tube, a drop forming pallet having an annular liquid chamber on an outer periphery of the center tube, an outer wall of the annular liquid chamber extending above the center tube, and having a solution pocket having an upper opening formed therein; A lower glass placed on the upper end of each central tube to close the central tube, and an upper glass placed on the upper opening of the solution pocket to close each solution pocket, and the grease applying device is provided for the central tube. The hanging drop for protein crystallization according to any one of claims 1 to 4, wherein grease is applied to an upper end opening, and the glass attaching / detaching device places the lower glass on the upper end opening of the central tube. Making equipment.

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