A serial crystallography experimental device based on chip technology and its assembly method

Abstract

The present invention discloses a serial crystallography experimental device based on chip technology and its assembly method. The device includes a chip, a chip base, a chip operation disk, a chip lower disk, a chip upper disk, and a low-background scattering film. The chip is provided with multiple hollow chip holes for supporting and adsorbing crystals. The chip lower disk and the chip upper disk are nested to fix the low-background scattering film to the upper and lower ends of the chip base. The chip operation disk is provided with a deliquescence chamber groove and a deliquescence chamber for removing redundant crystal solution from the chip. The deliquescence chamber groove is used to accommodate the chip base. The side wall of the deliquescence chamber is provided with an exhaust hole connected to the exhaust pipe of the exhaust system, generating a force on the crystal solution so that the redundant crystal solution is sucked to the other end of the chip hole. The chip operation disk is provided with an observation chamber groove and an observation chamber for observing the state of the crystal in the chip hole. The present invention can quickly assist the chip in removing excess crystal solution, and the chip can be quickly observed, assembled, and installed on the line station.

Description

Technical Field

[0001] The present invention relates to the field of protein crystal loading, and more particularly 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 crystallography stations 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 primarily include fixed-target and mobile-phase techniques. Fixed-target techniques involve no relative movement of the crystal relative to the crystal carrier during data acquisition, while mobile-phase techniques involve relative movement of the crystal relative to the crystal carrier during data acquisition. Compared to mobile-phase techniques, fixed-target techniques offer a higher crystal hit rate, greater sample utilization, and lower technical difficulty.

[0004] Conventional fixed-target technologies primarily include chip technology and membrane-based loading techniques. Chip technology has gained widespread attention at fourth-generation synchrotron radiation sources and free-electron laser facilities. The advantages of chip technology lie in the compatibility of the chip's microscale with the size of the crystal, and the ability to transfer a relatively large number of crystals at a time. However, the development of chip technology is still immature, and there are several problems with directly applying existing chip technology to serial crystallography. First, excess crystal solution can interfere with the process. Second, the crystals on the chip are difficult to efficiently transfer to the line station. Pre-processing of samples on the chip is cumbersome, and long-term data acquisition cannot be guaranteed. Therefore, serial crystallography based on chip technology needs to be continuously improved to adapt to new experimental requirements. Summary of the Invention

[0005] In order to solve the efficiency problem of existing chip technology, the present invention provides an experimental device and method based on chip-on-chip technology. By designing a chip supporting device, efficient serial crystallography experiments can be 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 supports the chip and, in conjunction with the chip manipulation tray, facilitates crystal desalting and observation. After sample preparation, the upper and lower chip trays nest together to secure the low-background scattering film to the ends of the chip base, effectively attaching the film to the chip assembly. The top and bottom chip caps thread together to secure the upper and lower chip trays. Finally, the bottom chip cap can be assembled with the goniometer head of a conventional crystallography line station to complete sample loading. The low-background scattering film serves two purposes: to slow the evaporation rate of the crystal solution and to minimize any interference with the quality of the crystal diffraction data.

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

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

[0010] Preferably, the upper and lower chip plates are each provided with a semi-threaded upper and lower plate at their ends, respectively. These semi-threaded upper and lower plate components can be joined to form a single threaded column. Furthermore, the bottom side of the lower chip plate has rounded corners to facilitate assembly of the low-background scattering film onto 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 ​​together. 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 by comprising a chip 10, a chip base 20, a chip operation tray 30, a chip lower tray 40, a chip upper tray 50 and a low background scattering film;

[0015] The chip 10 is provided with a plurality of hollow chip holes 11 for supporting and absorbing 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 deliquescence chamber groove 34 and a deliquescence chamber 32 for removing redundant crystal solution in the chip 10; wherein, the deliquescence chamber 32 is located below the deliquescence chamber groove 34 and is communicated with the deliquescence chamber groove 34, the deliquescence chamber groove 34 is used to accommodate the chip base 20, and a suction hole is provided on the side wall of the deliquescence 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;

[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, thereby obtaining 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 line station.

[0024] Furthermore, 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.

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

[0026] 1) Transferring the crystal solution to the chip 10, wherein the chip pores 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, 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 panel 30, and using the air extraction system to generate a force on the crystallite solution so that the redundant crystallite 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 assembled 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 sample 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 in removing 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 Schematic diagram of an experimental device for chip-on-chip technology according to a preferred embodiment of the present invention.

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

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

[0037] Figure 6 2 is a schematic structural diagram of a chip-on-disk according to a preferred embodiment of the present invention.

[0038] Figure 7 2 is a schematic structural diagram of a chip top cap according to a preferred embodiment of the present invention.

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

[0040] Figure 9 1 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 tray 30, a chip lower tray 40, and a chip upper tray 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 crystallography experiment, the crystal solution is first transferred to the chip 10 and evenly coated into the chip holes 11 within the area. The chip holes 11 are hollow, and they 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. Because crystallite solution interferes with crystal data acquisition, the chip base 20 needs to be placed in the deliquescence chamber groove 34 of the chip operation tray 30. The deliquescence chamber groove 34 is designed as a circular elongated rectangle, used to support the chip base 20 and the upper and lower chip trays, respectively. A certain degree of deliquescence can be performed in the deliquescence chamber 32. A conventional exhaust pipe is assembled with the air hole 35 of the chip operation tray 30. The chip base groove 21 in the center of the chip base 20 is a bottomless through-hole. After the chip base 20, equipped with the chip 10, is placed in the deliquescence chamber groove 34, there is no barrier between the chip well 11 on the chip 10 and the bottom of the deliquescence chamber 32. The air hole 35 is connected to the exhaust system, and the entire deliquescence chamber is in a sealed state. The exhaust system can exert force on the crystallite solution in the chip well 11, causing excess crystallite solution to be sucked to the other end of the chip well 11 and dissipated. Crystals larger than the chip well size will not be sucked to the lower end of the crystal well 11, so crystals with a small amount of crystallite solution will remain at the upper end of the chip well 11.

[0045] After the liquid is removed, the chip base 20 is placed in the observation chamber 31 of the chip operation tray 30. The bottom of the observation chamber 31 is hollow. After placement, the state of the crystals in the chip hole 11 can be observed under a microscope to ensure that the crystals are intact and there is no large overlap.

[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 panel 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. Then the chip lower plate 40 is taken out and the film is pressed down from above 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 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, thereby completing the sealing of one side of the chip base 20. The chip base 20 and chip lower plate 40 assembly is then flipped over so that the unsealed side of the chip base 20 faces upward. The low-background scattering film is again applied over the observation chamber 31 to completely cover it. Finally, the chip upper plate 50 is placed over the observation chamber, and the two upper plate semi-threaded posts 51 of the chip upper plate 50 are aligned with the two lower plate semi-threaded posts 41 of the chip lower plate 40. After alignment, the upper plate semi-threaded posts 51 and the lower plate semi-threaded posts 41 form a complete threaded post. The assembly of the device is then completed by aligning the top cap internal threads 61 of the chip top cap 60 and the bottom cap internal threads 71 ​​of the chip bottom cap 70 with the two threaded posts formed at the upper and lower ends, respectively. The chip bottom cap 70 also has a bottom cap hole 72 with a built-in magnet that can be assembled with the goniometer head of a conventional crystallography line station. Once assembled, chip-based serial crystallography experiments can be performed.

[0047] The above description is merely a preferred embodiment of the present invention and is not intended to limit the scope of the present invention. Various modifications are possible. In other words, any simple, equivalent changes and modifications made in accordance with the claims and description of the present invention are within the scope of protection of the patent claims. Anything not fully described in this invention constitutes conventional technical content.

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 the 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 deliquescence chamber groove (34) and a deliquescence chamber (32), which are used to remove redundant crystal solutions in the chip (10); wherein the deliquescence chamber (32) is located below the deliquescence chamber groove (34) and is communicated with the deliquescence chamber groove (34), the deliquescence chamber groove (34) is used to accommodate the chip base (20), and a suction hole is provided on the side wall of the deliquescence 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); 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 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), thereby obtaining the sample 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).

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 sample 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 line station.

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), then placing the chip base (20) on the deliquescence chamber groove (34), connecting the exhaust hole on the side wall of the deliquescence chamber (32) with the air hole (35) of the chip operation disk (30), and generating a force on 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 assembled 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 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), thereby obtaining the sample 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).

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 sample 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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