Serial crystallography experiment device based on chip technology and assembling method thereof

Abstract

The invention discloses a serial crystallography experiment device based on a chip technology and an assembling method thereof. The device comprises a chip, a chip base, a chip operation board, a chip lower disc, a chip upper disc and a low background scattering film, a plurality of hollow chip holes are formed in the chip and are used for supporting and adsorbing the crystal; the lower chip disc and the upper chip disc fix the low background scattering films at the upper end and the lower end of the chip base in a nesting manner; a liquid removal chamber groove and a liquid removal chamber are arranged on the chip operation board and are used for removing redundant crystal solution in the chip; the groove of the liquid removal chamber is used for accommodating a chip base, an air exhaust hole is formed in the side wall of the liquid removal chamber and connected with an air exhaust pipe of an air exhaust system, and acting force is generated on the crystal solution to enable the redundant crystal solution to be sucked to the other end of a chip hole; and the chip operation board is provided with an observation chamber groove and an observation chamber and is used for observing the state of the crystal in the chip hole. According to the invention, the chip can be rapidly assisted to remove redundant crystal solution, and the chip can be rapidly observed, assembled and installed on a line station.

Description

Technical Field

[0001] The present invention relates to the field of protein crystal loading, and more specifically to an experimental device and method based on chip loading technology. Background Art

[0002] In recent years, serial crystallography has gradually become a popular research direction in crystallographic lines around the world. Serial crystallography includes high-throughput preparation, loading and data processing technologies of crystals, among which high-throughput loading of crystals is the key and difficult point in the entire serial crystallography.

[0003] Existing serial crystallography techniques mainly include fixed target technology and mobile phase technology. Fixed target technology means that the crystal does not move relative to the crystal carrier during data collection, while mobile phase technology means that the crystal moves relative to the crystal carrier during data collection. Compared with mobile phase technology, the characteristics of fixed target technology are mainly high crystal hit rate, high sample utilization rate, and low technical implementation difficulty.

[0004] Conventional fixed target technologies mainly include chip technology and membrane-based loading technology. Among them, chip technology has been widely recognized in the fourth-generation synchrotron radiation source and free electron laser device. The advantage of chip technology is that the micro size of the chip is more compatible with the size of the crystal, and the number of crystals that can be transferred by the chip at one time is relatively objective. However, the development of chip technology is still in an immature stage. There are some problems when the existing chip technology is directly applied to serial crystallography: first, there is a certain interference from the excess crystal solution. Secondly, the crystals on the chip are difficult to transfer to the line station efficiently with the chip. The pretreatment of samples on the chip is more troublesome, and it is impossible to guarantee that the crystals can collect data for a long time. Therefore, serial crystallography based on chip technology needs to be continuously improved to adapt to new experimental needs. Summary of the invention

[0005] In order to solve the efficiency problem of the existing chip technology, the present invention provides an experimental device and method based on chip loading technology. By designing a chip matching device, an efficient serial crystallography experiment is carried out.

[0006] The chip loading experiment device according to the present invention mainly comprises a chip, a chip base, a chip upper plate, a chip lower plate, a chip operation plate, a chip bottom cap and a chip top cap.

[0007] The chip base can support the chip and be assembled with the chip operation plate to complete the liquid removal and observation of the crystal. After the sample preparation is completed, the chip upper plate and the chip lower plate can be nested to fix the low background scattering film on both ends of the chip base, so that the low background scattering film can be efficiently assembled on the chip device. The chip top cap and the chip bottom cap can be fixed to the chip upper plate and the chip lower plate by threaded assembly. Finally, the chip bottom cap can be assembled with the goniometer head of the conventional crystallography line station to complete the sample loading. The low background scattering film has two functions: one is to reduce the volatilization rate of the crystal solution, and the other is to reduce the interference of the film on the quality of the crystal diffraction data.

[0008] Preferably, the chip base has a built-in chip groove to assemble the chip without blocking the chip hole.

[0009] Preferably, the chip operation panel has an observation chamber and a deliquidation chamber, and the two chambers are respectively provided with an observation chamber groove and a deliquidation chamber groove to place the chip or chip base. The observation chamber is hollowed out to observe the crystal state, and the deliquidation chamber is non-hollowed out. The side wall of the deliquidation chamber is provided with an exhaust hole, which can be connected to the suction device.

[0010] Preferably, the upper plate and the lower plate are provided with upper plate semi-threaded columns and lower plate semi-threaded columns at both ends, respectively, and the semi-threaded columns at both ends of the upper plate and the lower plate can be spliced ​​into a whole threaded column. Secondly, the bottom side of the lower plate is provided with rounded corners to facilitate the assembly of the low background scattering film on the chip base.

[0011] Preferably, a top cap thread groove is provided in the chip top cap, and the top cap thread groove can be assembled with the entire thread column at any end after the upper and lower plates of the chip are spliced.

[0012] Preferably, a bottom cap thread groove and a bottom cap hole are provided in the chip bottom cap, the bottom cap thread groove can be assembled with the entire thread column at either end after the upper and lower plates of the chip are spliced, and the bottom cap hole has a built-in magnetic attraction and can be assembled with the goniometer head of a conventional crystallographic line station.

[0013] The technical solution of the present invention is:

[0014] A serial crystallography experimental device based on chip technology, characterized in that it comprises a chip 10, a chip base 20, a chip operation plate 30, a chip lower plate 40, a chip upper plate 50 and a low background scattering film;

[0015] The chip 10 is provided with a plurality of hollow chip holes 11 for supporting and adsorbing crystals in the crystal solution transferred to the chip 10;

[0016] The chip base 20 is provided with a chip base groove 21 for placing the chip 10;

[0017] The chip lower plate 40 and the chip upper plate 50 are fixed to the upper and lower ends of the chip base 20 by nesting to obtain a sample loading unit;

[0018] The chip operation disk 30 is provided with a dehydration chamber groove 34 and a dehydration chamber 32, which are used to remove redundant crystal solutions in the chip 10; wherein the dehydration chamber 32 is located below the dehydration chamber groove 34 and is communicated with the dehydration chamber groove 34, the dehydration chamber groove 34 is used to accommodate the chip base 20, and a suction hole is provided on the side wall of the dehydration chamber 32, which is used to communicate with the air hole 35 of the chip operation disk 30; the air hole 35 is used to be connected to the suction pipe of the suction system, and the suction system generates a force on the crystal solution so that the redundant crystal solution in the chip 10 is sucked to the other end of the chip hole 11;

[0019] The chip operation disk 30 is provided with an observation chamber groove 33 and an observation chamber 31 for observing the state of the crystal in the chip hole 11; the observation chamber 31 is located below the observation chamber groove 33 and is connected to the observation chamber groove 33, and the observation chamber groove 33 is used to accommodate the chip base 20.

[0020] Furthermore, the chip lower plate 40 and the chip upper plate 50 are annular structures, and two lower plate semi-threaded columns 41 are provided on the side wall of the annular structure of the chip lower plate 40; two upper plate semi-threaded columns 51 are provided on the side wall of the annular structure of the chip upper plate 50; the annular structure of the chip lower plate 40 and the chip upper plate 50 is used to clamp the chip base 20 on which the chip 10 is placed, and the two lower plate semi-threaded columns 41 and the two upper plate semi-threaded columns 51 correspond to each other in position to form two threaded columns, one of which is fixed by the top cap internal thread 61 of a chip top cap 60, and the other threaded column is fixed by the bottom cap internal thread 71 of a chip bottom cap 70, so as to obtain the loading unit; a low background scattering film is respectively provided between the chip lower plate 40, the chip upper plate 50 and the chip base 20 to cover the chip 10.

[0021] Furthermore, the bottom of the deliquidation chamber 32 is a closed structure, and the deliquidation chamber groove 34 includes a hollow annular groove and two rectangular grooves, the hollow part of the annular groove is connected to the deliquidation chamber 32, and the rectangular groove is connected to the annular groove; wherein, the annular groove is matched and assembled with the annular structure of the chip lower plate 40 and the chip upper plate 50, and the rectangular groove is matched and assembled with the threaded column to obtain the loading unit.

[0022] Furthermore, the bottom of the observation chamber 31 is a hollow structure, and the observation chamber groove 33 includes a hollow annular groove and two rectangular grooves, the hollow part of the annular groove is connected to the observation chamber 31, and the rectangular groove is connected to the annular groove; the annular groove is matched and assembled with the annular structure of the chip lower plate 40 and the chip upper plate 50; the rectangular groove is matched with the threaded column for accommodating the threaded column.

[0023] Furthermore, the chip bottom cap 70 has a bottom cap hole 72 , and the bottom cap hole 72 has a built-in magnetic attraction for assembly with a goniometer head of a crystallographic beamline.

[0024] Furthermore, a rounded corner 42 is provided on the annular structure of the chip bottom plate 40 for positioning and assembling with the chip base 20 ; and the bottom of the chip base groove 21 is a through-hole structure.

[0025] A method for assembling a serial crystallography experimental device, the steps comprising:

[0026] 1) transferring the crystal solution to the chip 10, wherein the chip holes of the chip 10 are hollow structures for supporting and adsorbing the crystals;

[0027] 2) placing the chip 10 in the chip base groove 21 in the middle of the chip base 20, and then placing the chip base 20 on the deliquescence chamber groove 34, connecting the air extraction holes on the side wall of the deliquescence chamber 32 with the air holes 35 of the chip operation disk 30, and applying force to the crystal solution through the air extraction system so that the redundant crystal solution in the chip 10 is sucked to the other end of the chip hole 11;

[0028] 3) placing the chip base 20 processed in step 2) on the observation chamber groove 33 for observation;

[0029] 4) Cover the upper end surface of the chip base 20 with a low-background scattering film, press the low-background scattering film down with the chip lower plate 40 and assemble it with the chip base 20 to complete the sealing of the upper end surface of the chip base 20; then flip the assembly structure of the chip base 20 and the chip lower plate 40, cover the lower end surface of the chip base 20 with a low-background scattering film, press the low-background scattering film down with the chip upper plate 50 and assemble it with the chip base 20 to complete the sealing of the lower end surface of the chip base 20, and obtain a loading unit.

[0030] Compared with the prior art, the present invention has the following positive effects:

[0031] The supporting device of the present invention can quickly assist the chip to remove the interference of excess crystal solution. At the same time, the chip can be quickly observed, assembled and installed on the line station, and the crystal state on the chip can be maintained for a long time and effective data collection can be completed. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 It is a schematic diagram of an experimental device for chip-loading technology according to a preferred embodiment of the present invention.

[0033] Figure 2 It is a schematic diagram of the structure of a chip according to a preferred embodiment of the present invention.

[0034] Figure 3 It is a schematic structural diagram of a chip base according to a preferred embodiment of the present invention.

[0035] Figure 4 It is a schematic diagram of the structure of a chip operating disk according to a preferred embodiment of the present invention.

[0036] Figure 5 It is a schematic structural diagram of a chip lower plate according to a preferred embodiment of the present invention.

[0037] Figure 6 It is a schematic structural diagram of a chip-on-disk according to a preferred embodiment of the present invention.

[0038] Figure 7 Schematic diagram of the structure of a chip top cap according to a preferred embodiment of the present invention.

[0039] Figure 8 It is a schematic diagram of the assembly end structure of a chip bottom cap according to a preferred embodiment of the present invention.

[0040] Fig. 9 It is a schematic diagram of the fixed end structure of a chip bottom cap according to a preferred embodiment of the present invention. DETAILED DESCRIPTION

[0041] The preferred embodiments of the present invention are given below in conjunction with the accompanying drawings and described in detail.

[0042] Example

[0043] like Figure 1 As shown, the serial crystallography experimental device based on chip technology mainly includes a chip 10, a chip base 20, a chip operation plate 30, a chip lower plate 40, and a chip upper plate 50. In addition, there are also assembly devices such as a chip top cap 60 and a chip bottom cap 70.

[0044] When conducting a crystallographic experiment, the crystal solution is first transferred to the chip 10 and evenly coated into the chip holes 11 in the area. The chip holes 11 are hollow and mainly support and adsorb the crystals through hollow holes that are smaller than the size of the crystals. Then the chip 10 is placed in the chip base groove 21 in the middle of the chip base 20. Due to the interference of crystal solution on crystal data collection, the chip base 20 needs to be placed on the dehydration chamber groove 34 of the chip operation disk 30. The dehydration chamber groove 34 is designed as a ring-shaped and elongated rectangle, which is used to support the chip base 20 and the upper and lower plates of the chip respectively. A certain degree of dehydration can be performed in the dehydration chamber 32, and a conventional exhaust pipe is assembled with the air hole 35 of the chip operation disk 30; the chip base groove 21 in the middle of the chip base 20 is a bottomless through hole. After the chip base 20 equipped with the chip 10 is placed on the dehydration chamber groove 34, there is no barrier between the chip hole 11 on the chip 10 and the bottom of the dehydration chamber 32. The air hole 35 is connected to the exhaust system, and the entire dehydration chamber is in a closed state. The exhaust system can generate a force on the crystal solution in the chip hole 11, so that the redundant crystal solution is sucked to the other end of the chip hole 11 and dissipated. Crystals with a crystal size larger than the chip hole size will not be sucked to the lower end of the crystal hole 11, so the upper end of the chip hole 11 can leave crystals with a small amount of crystal solution.

[0045] After the liquid is removed, the chip base 20 is placed in the observation chamber 31 of the chip operation disk 30. The bottom of the observation chamber 31 is hollow. After placement, the state of the crystal in the chip hole 11 can be observed under a microscope to ensure that the crystal is intact and does not overlap to a large extent.

[0046] After the crystal preparation is completed, the chip loading device is assembled to obtain the loading unit. The assembly process can be completed through the observation chamber 31 or the liquid removal chamber 32 of the chip operation disk 30, or it can be self-assembled outside the device. The steps are the same when assembling using the observation chamber 31 or the liquid removal chamber 32. Take the observation chamber 31 as an example: after the chip base 20 is placed in the observation chamber 31, the low background scattering film is laid on the top of the observation chamber 31 to completely cover it, and then the chip lower plate 40 is taken out and the film is pressed down from the top of the observation chamber 31 and assembled with the chip base 20. The rounded corners 42 of the chip lower plate 40 mainly function to facilitate the positioning and assembly with the chip base 20. After assembly, the lower plate semi-threaded column 41 of the chip lower plate 40 is just placed on the observation chamber groove 33, and the sealing of one side of the chip base 20 is completed. Afterwards, the assembly device of the chip base 20 and the chip lower plate 40 is turned over so that the unsealed side of the chip base 20 faces upward, and the low background scattering film is laid again on the observation chamber 31 to completely cover it. Finally, the chip upper plate 50 is placed above the observation chamber, and the two upper plate semi-threaded columns 51 of the chip upper plate 50 are aligned with the two lower plate semi-threaded columns 41 of the chip lower plate 40. After alignment, the upper plate semi-threaded columns 51 and the lower plate semi-threaded columns 41 form a complete threaded column, and then the top cap internal thread 61 of the chip top cap 60 and the bottom cap internal thread 71 of the chip bottom cap 70 are respectively assembled with the two threaded columns formed at the upper and lower ends to complete the assembly of the device. The chip bottom cap 70 also has a bottom cap hole 72, which has a built-in magnetic attraction and can be assembled with the angle measuring head of a conventional crystallographic line station. After the assembly is completed, a chip-based serial crystallography experiment can be carried out.

[0047] The above is only a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention. The above embodiment of the present invention can also be modified in various ways. That is, all simple, equivalent changes and modifications made according to the claims and the description of the present invention fall within the scope of protection of the claims of the present invention. The contents not described in detail in the present invention are all conventional technical contents.

Claims

1. A serial crystallography experimental device based on chip technology, characterized in that: It comprises a chip (10), a chip base (20), a chip operation plate (30), a chip lower plate (40), a chip upper plate (50) and a low background scattering film; The chip (10) is provided with a plurality of hollow chip holes (11) for supporting and adsorbing crystals in a crystal solution transferred to the chip (10); The chip base (20) is provided with a chip base groove (21) for placing the chip (10); The chip lower plate (40) and the chip upper plate (50) are nested to fix the low background scattering film at the upper and lower ends of the chip base (20) to obtain a sample loading unit; The chip operation disk (30) is provided with a dehydration chamber groove (34) and a dehydration chamber (32) for removing redundant crystal solution in the chip (10); wherein the dehydration chamber (32) is located below the dehydration chamber groove (34) and is communicated with the dehydration chamber groove (34); the dehydration chamber groove (34) is used to accommodate the chip base (20); a suction hole is provided on the side wall of the dehydration chamber (32) for communicating with the air hole (35) of the chip operation disk (30); the air hole (35) is used to be connected to the suction pipe of the suction system, and the suction system generates a force on the crystal solution so that the redundant crystal solution in the chip (10) is sucked to the other end of the chip hole (11); The chip operation disk (30) is provided with an observation chamber groove (33) and an observation chamber (31) for observing the state of the crystal in the chip hole (11); the observation chamber (31) is located below the observation chamber groove (33) and is connected to the observation chamber groove (33); the observation chamber groove (33) is used to accommodate the chip base (20).

2. The serial crystallography experimental device according to claim 1, characterized in that: The chip lower plate (40) and the chip upper plate (50) are annular structures, and two lower plate semi-threaded columns (41) are arranged on the side wall of the annular structure of the chip lower plate (40); two upper plate semi-threaded columns (51) are arranged on the side wall of the annular structure of the chip upper plate (50); the annular structures of the chip lower plate (40) and the chip upper plate (50) are used to clamp the chip base (20) on which the chip (10) is placed, and the two lower plate semi-threaded columns (41) and the two upper plate semi-threaded columns (51) correspond to each other in position to form two threaded columns, one of which is fixed by the top cap internal thread (61) of a chip top cap (60), and the other is fixed by the bottom cap internal thread (71) of a chip bottom cap (70), so as to obtain the loading unit; a low background scattering film is respectively arranged between the chip lower plate (40), the chip upper plate (50) and the chip base (20) to cover the chip (10).

3. The serial crystallography experimental device according to claim 2, characterized in that: The bottom of the deliquidation chamber (32) is a closed structure, and the deliquidation chamber groove (34) includes a hollow annular groove and two rectangular grooves, the hollow part of the annular groove is connected to the deliquidation chamber (32), and the rectangular groove is connected to the annular groove; wherein, the loading unit is obtained by matching and assembling the annular structure of the chip lower plate (40) and the chip upper plate (50), and matching and assembling the rectangular groove with the threaded column.

4. The serial crystallography experimental device according to claim 2, characterized in that: The bottom of the observation chamber (31) is a hollow structure, and the observation chamber groove (33) includes a hollow annular groove and two rectangular grooves. The hollow part of the annular groove is connected to the observation chamber (31), and the rectangular groove is connected to the annular groove; the annular groove is matched and assembled with the annular structure of the chip lower plate (40) and the chip upper plate (50); the rectangular groove is matched with the threaded column and is used to accommodate the threaded column.

5. The serial crystallography experimental device according to claim 2, characterized in that: The chip bottom cap (70) is provided with a bottom cap hole (72), and the bottom cap hole (72) has a built-in magnetic attraction for assembly with a goniometer head of a crystallographic beamline.

6. The serial crystallography experimental device according to any one of claims 1 to 5, characterized in that: The annular structure of the chip bottom plate (40) is provided with a rounded corner (42) for positioning and assembling with the chip base (20); the bottom of the chip base groove (21) is a through-hole structure.

7. A method for assembling the serial crystallography experimental device according to claim 1, comprising the steps of: 1) transferring the crystal solution into a chip (10), wherein the chip holes of the chip (10) are hollow structures for supporting and adsorbing crystals; 2) placing the chip (10) in the chip base groove (21) in the middle of the chip base (20), and then placing the chip base (20) on the dehydration chamber groove (34), connecting the exhaust hole on the side wall of the dehydration chamber (32) with the air hole (35) of the chip operation disk (30), and applying force to the crystal solution through the exhaust system so that the redundant crystal solution in the chip (10) is sucked to the other end of the chip hole (11); 3) placing the chip base (20) processed in step 2) on the observation chamber groove (33) for observation; 4) using a low-background scattering film to cover the upper end surface of the chip base (20), using a chip lower plate (40) to press down the low-background scattering film and assemble it with the chip base (20), thereby completing the sealing of the upper end surface of the chip base (20); then turning over the assembly structure of the chip base (20) and the chip lower plate (40), using a low-background scattering film to cover the lower end surface of the chip base (20), using a chip upper plate (50) to press down the low-background scattering film and assemble it with the chip base (20), thereby completing the sealing of the lower end surface of the chip base (20), thereby obtaining a sample loading unit.

8. The method according to claim 7, characterized in that The chip lower plate (40) and the chip upper plate (50) are annular structures, and two lower plate semi-threaded columns (41) are arranged on the side wall of the annular structure of the chip lower plate (40); two upper plate semi-threaded columns (51) are arranged on the side wall of the annular structure of the chip upper plate (50); the annular structures of the chip lower plate (40) and the chip upper plate (50) are used to clamp the chip base (20) on which the chip (10) is placed, and the two lower plate semi-threaded columns (41) and the two upper plate semi-threaded columns (51) correspond to each other in position to form two threaded columns, one of which is fixed by the top cap internal thread (61) of a chip top cap (60), and the other is fixed by the bottom cap internal thread (71) of a chip bottom cap (70), so as to obtain the loading unit; a low background scattering film is respectively arranged between the chip lower plate (40), the chip upper plate (50) and the chip base (20) to cover the chip (10).

9. The method according to claim 8, characterized in that The bottom of the deliquidation chamber (32) is a closed structure, and the deliquidation chamber groove (34) includes a hollow annular groove and two rectangular grooves, the hollow part of the annular groove is connected to the deliquidation chamber (32), and the rectangular groove is connected to the annular groove; wherein, the loading unit is obtained by matching and assembling the annular structure of the chip lower plate (40) and the chip upper plate (50), and matching and assembling the rectangular groove with the threaded column.

10. The method according to claim 8, characterized in that The bottom of the observation chamber (31) is a hollow structure, and the observation chamber groove (33) includes a hollow annular groove and two rectangular grooves. The hollow part of the annular groove is connected to the observation chamber (31), and the rectangular groove is connected to the annular groove; the annular groove is matched and assembled with the annular structure of the chip lower plate (40) and the chip upper plate (50); the rectangular groove is matched with the threaded column and is used to accommodate the threaded column.

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