Automatic freezing electron microscope sample preparation function enhancing device

Abstract

The invention provides an automatic cryoelectron microscope sample preparation function enhancing device which comprises a transversely-arranged first linear module, the first linear module is provided with a movable first sliding plate, the first sliding plate is provided with a rotary table device, the rotary table device is provided with a rotatable rotating flange plate, and the rotating flange plate is provided with a net carrying box low-temperature container. A third linear module which is vertically arranged is arranged above the first linear module, the third linear module is provided with a movable third sliding plate, the third sliding plate is provided with a rotatable lower extension rod assembly, the lower end of the lower extension rod assembly is provided with a pair of sample loading tweezers and a liquid suction clamp, the upper end of the liquid suction clamp is connected with the third linear module, and the lower end of the liquid suction clamp is connected with the second linear module. A fourth linear module is transversely arranged on one side of the third linear module and provided with a movable fourth sliding plate, a fixing clamp is arranged on the fourth sliding plate, a quantitative pipette is arranged at the position of the fixing clamp, and a linear ejection cylinder is arranged at the rear end of the quantitative pipette on the fourth sliding plate. And the success rate is low.

Description

Technical Field

[0001] This invention relates to the field of electron microscopy sample preparation, and in particular to an automated cryo-electron microscopy sample preparation function enhancement device. Background Technology

[0002] Cryo-electron microscopy (cryo-EM) grids are a key component in cryo-EM sample preparation. They serve as scaffolds or trays to support biological samples such as viruses and proteins, and their quality and characteristics directly affect the final imaging results.

[0003] Before the sample is placed on the grid, it is usually pre-cooled in a liquid nitrogen environment. Then, the sample is added dropwise, and excess water is absorbed by filter paper. Then, it is placed in liquid ethane to freeze into a layer of glassy ice. Finally, it is quickly placed back into the grid storage box in a liquid nitrogen environment for subsequent testing.

[0004] Due to the small size of the sample carrier, the traditional process mainly involves manually handling the carrier with tweezers for transfer. Since the carrier needs to stand for several minutes to tens of minutes after sample loading, it is difficult to maintain a handheld position for extended periods. Furthermore, excess moisture is usually absorbed after sample loading, and the handheld filter paper can easily touch the carrier surface, damaging the sample layer. Therefore, there is an urgent need for an automated device to replace manual sample preparation. Summary of the Invention

[0005] This invention provides an automated cryo-electron microscopy sample preparation function enhancement device, which solves the problems of high labor intensity and low success rate of manual sample preparation in cryo-electron microscopy.

[0006] To solve the above-mentioned technical problems, the technical solution adopted by the present invention is: an automated cryo-electron microscopy sample preparation function enhancement device, comprising a first linear module arranged horizontally, the first linear module having a movable first sliding plate, a turntable device having a rotatable rotating flange having a low-temperature container for carrying a mesh tray on the rotating flange, a third linear module arranged vertically above the first linear module having a movable third sliding plate, a rotatable lower extension rod assembly having a sample loading forceps at the lower end of the lower extension rod assembly, and a liquid suction clamp having its upper end connected to the third linear module, a fourth linear module arranged horizontally on one side of the third linear module having a movable fourth sliding plate, a fixing clamp having a fixed clamp having a quantitative pipette at the fixed clamp, and a linear top cylinder having a linear top cylinder at the rear end of the quantitative pipette on the fourth linear module.

[0007] In a preferred embodiment, the low-temperature container for the wire mesh carrier includes a first holding container, an annular groove inside the first holding container, a wire mesh carrier placement rack inside the annular groove, a plurality of wire mesh carrier positioning slots along the circumference of the wire mesh carrier placement rack, a wire mesh carrier in each wire mesh carrier positioning slot, a plurality of wire mesh slots along the circumference of the wire mesh carrier, the wire mesh slots being used to place the sample wire mesh, and a partition groove in the center of the first holding container, a second holding container inside the partition groove.

[0008] In a preferred embodiment, the lower extension rod assembly includes a core rod, the lower end of which is provided with a detachable connecting rod. The sample tweezers are connected to the lower end of the connecting rod. A rotatable rotating seat is provided on the third sliding plate, and an end fixing seat is provided on the rotating seat. The upper end of the core rod is sleeved with the end fixing seat.

[0009] In a preferred embodiment, the lower extension rod assembly further includes an outer sleeve sleeved around the outside of the core rod. The outer sleeve is threadedly connected to the core rod. A rotatable guide sleeve is also provided below the rotating seat on the third sliding plate. A straight guide groove is provided circumferentially on the inner wall of the guide sleeve. A guide strip is provided on the outer arm of the outer sleeve. The guide strip is slidably engaged with the straight guide groove. A locking sleeve is provided at the lower end of the outer sleeve. The lower end of the locking sleeve presses the forceps arm of the sample loading forceps to clamp the forceps arm.

[0010] In a preferred embodiment, the upper end of the locking sleeve is provided with a sliding part, and the outer wall of the sliding part is provided with an annular locking groove. The lower end of the outer sleeve is also provided with a connecting sleeve. The lower end of the connecting sleeve has multiple deformable arms along the circumference. The inner side of the deformable arms is provided with a locking protrusion. The locking protrusion slides and engages with the annular locking groove so that the locking sleeve can rotate relative to the connecting sleeve. The lower end of the locking sleeve is provided with a flexible bushing. The outer side of the sample forceps arm is provided with a protrusion. The flexible bushing engages with the protrusion. The outer side of the deformable arm is covered with a limit sleeve.

[0011] In a preferred embodiment, the liquid suction clamp includes two clamping flaps arranged opposite each other. A filter paper is provided on one side of each clamping flap, and a connecting arm is provided on the other side of each clamping flap. The upper end of the connecting arm is provided with an inner bend. A hinge seat and a connecting seat are provided on the base frame. The middle part of the connecting arm is hinged to the hinge seat. A tension spring is connected to the connecting seat. A connecting ear is provided at the lower end of the connecting arm. The tension spring holds the connecting ear. A lifting rod is connected to the lower end of the third sliding plate. A fork-shaped block is provided at the lower end of the lifting rod. A wedge-shaped surface is provided on the inner side of the fork-shaped block. The wedge-shaped surface presses the inner bend to make the clamping flaps swing.

[0012] In a preferred embodiment, the clamping flap includes an end cap and a rear housing. The rear housing is connected to the connecting arm. A movable sleeve that can reciprocate is provided inside the rear housing. One end of the movable sleeve has a front ring tooth, and the other end of the movable sleeve has a rear ring tooth. The end cap near the movable sleeve has a front ring tooth, which meshes with the front ring tooth. A rear fixed seat is provided on the base. The end of the rear fixed seat near the rear housing has a rear ring tooth, which meshes with the rear ring tooth. A rotating disk is also provided. One end of the rotating disk passes through the end cap to connect with the movable sleeve. The end face of the rotating disk has a deformable flexible ring. The filter paper is placed on the flexible ring. The front ring tooth, the front ring tooth, the rear ring tooth, and the rear ring tooth are inclined teeth. The meshing of the rear ring tooth with the rear ring tooth and the meshing of the front ring tooth with the front ring tooth both cause the rotating disk to rotate circumferentially in the same direction.

[0013] In the preferred embodiment, a magnetic ring is provided on one side of the movable sleeve, and multiple electromagnets are provided circumferentially on the upper part of the rear housing. When the electromagnets are energized, they generate a repulsive force on the magnetic ring, which can push the movable sleeve close to the end cover so that the front ring teeth mesh with the front end ring teeth. When the electromagnets are de-energized, the movable sleeve is reset under the action of the tension spring so that the rear ring teeth mesh with the rear end ring teeth.

[0014] In the preferred embodiment, the end fixing seat is provided with a rotatable ball joint sleeve that is threadedly connected to the core rod. The side wall of the end fixing seat is provided with multiple threaded tightening screws along the circumferential direction, and one end of each tightening screw abuts against the outer wall of the core rod.

[0015] In the preferred embodiment, a lower bushing is provided at the lower end of the outer sleeve, and the lower bushing is sleeved with the core rod.

[0016] The beneficial effects of this invention are as follows: The use of an automated linear movement module in conjunction with an actuator replaces manual labor in complex operations such as transferring the mesh carrier position, adding samples, and aspirating liquid, ensuring sample consistency, reducing labor intensity, and improving sample quality. The mesh carrier low-temperature container can integrate multiple mesh carriers and other low-temperature auxiliary reagents. Through the coordination of rotation and movement mechanisms, it can achieve automatic mesh carrier gripping and transfer based on tweezers. The tweezers can be mounted on the automatic mechanism or separated for manual operation, making it more flexible to use. The use of a liquid aspiration clamp with switchable circumferential angles allows the filter paper to rotate by one circumferential angle each time it is used, ensuring that a single filter paper can be used multiple times by switching the contact area with the mesh carrier. This avoids cross-contamination of samples, prevents low filter paper utilization, and reduces the number of times filter paper needs to be manually pasted. Attached Figure Description

[0017] The present invention will be further described below with reference to the accompanying drawings and embodiments.

[0018] Figure 1 This is a schematic diagram of the mechanism arrangement of the present invention.

[0019] Figure 2 This is a side view of the present invention.

[0020] Figure 3 This is a diagram of the liquid suction clamp mechanism.

[0021] Figure 4 This is a schematic diagram of the suction clamp closing.

[0022] Figure 5 This is a top view of the fork-shaped block.

[0023] Figure 6 This is a schematic diagram of the lower extension rod assembly.

[0024] Figure 7 This is a partial view of the upper part of the lower extension rod assembly.

[0025] Figure 8 This is a cross-sectional view of the lower extension rod assembly.

[0026] Figure 9 This is a cross-sectional view of the sample loading forceps and locking sleeve.

[0027] Figure 10 This is a distribution diagram of the deformation arms.

[0028] Figure 11 It is a cross-sectional view of the flap.

[0029] Figure 12 This is a schematic diagram demonstrating the equivalent displacement of the movable sleeve teeth when straightened.

[0030] Figure 13 This is a schematic diagram showing the contact between the filter paper and the sample carrier mesh.

[0031] Figure 14 This is a schematic diagram of a cryogenic container with a wire mesh casing.

[0032] Figure 15 This is an exploded view of the first container.

[0033] Figure 16 This is an exploded view of the carrier box.

[0034] In the diagram: 101 for the wire mesh inlet; 102 for the sample inlet; 103 for the first upright; 104 for the second upright; 2 for the first linear module; 201 for the first sliding plate; 3 for the turntable device; 301 for the rotating flange; 5 for the wire mesh box cryogenic container; 501 for the sample wire mesh; 502 for the first container; 503 for the wire mesh box placement rack; 504 for the wire mesh box positioning groove; 505 for the wire mesh box; 506 for the wire mesh slot; 507 for the partition; 507 for the second container. 508; Annular groove; 509; Third linear module; 7; Third sliding plate; 701; Lower extension rod assembly; 702; Core rod; 703; Outer sleeve; 704; Guide sleeve; 705; Bearing seat; 706; Guide bar; 707; Linear guide groove; 708; Rotating seat; 709; End fixing seat; 710; Locking sleeve; 711; Flexible bushing; 712; Sliding part; 713; Annular snap-fit ​​groove; 714; Connecting sleeve; 715; Deformation arm; 716; Snap-fit ​​protrusion; 7 17; Limiting sleeve 718; Connecting rod 719; Ball joint sleeve 720; Locking nut 721; Tightening screw 722; First drive motor 723; Second drive motor 724; Lower bushing 725; Liquid suction clamp 8; Clamping disc 801; Connecting arm 802; Inner bend 803; Lifting rod 804; Fork block 805; Wedge surface 806; Hinge seat 807; Connecting seat 808; Tension spring 809; Connecting ear 810; Filter paper 8 11; Flexible ring 812; Rotating disk 813; End cap 814; Rear housing 815; Front ring tooth 816; Movable sleeve 817; Front ring tooth 818; Magnetic ring 819; Electromagnet 820; Rear ring tooth 821; Rear fixing seat 822; Rear ring tooth 823; Sample loading forceps 9; Protrusion 901; Fourth linear module 10; Fourth sliding plate 1001; Fixing clamp 1002; Quantitative pipette 11; Linear top cylinder 12. Detailed Implementation

[0035] like Figure 1-16 A device for enhancing the automated cryo-electron microscopy sample preparation function includes a horizontally arranged first linear module 2, which has a movable first sliding plate 201. A turntable device 3 is mounted on the first sliding plate 201, and a rotatable rotating flange 301 is mounted on the rotating flange 301. A low-temperature container 5 carrying a mesh tray is mounted on the rotating flange 301. Above the first linear module 2, a vertically arranged third linear module 7 is arranged, which has a movable third sliding plate 701. The third sliding plate 701 is equipped with... The device includes a rotatable lower extension rod assembly 702, with sample loading tweezers 9 at the lower end and a liquid suction clamp 8. The upper end of the liquid suction clamp 8 is connected to the third linear module 7. A fourth linear module 10 is arranged horizontally on one side of the third linear module 7. The fourth linear module 10 has a movable fourth sliding plate 1001. A fixing clamp 1002 is provided on the fourth sliding plate 1001. A quantitative pipette 11 is provided at the fixing clamp 1002. A linear top cylinder 12 is provided at the rear end of the quantitative pipette 11 on the fourth sliding plate 1001.

[0036] Each linear module adopts a servo motor-lead screw-slide rail structure.

[0037] The quantitative pipette 11 retains the push-button type pipette that allows for setting the pressure volume when operated manually.

[0038] In a preferred embodiment, the low-temperature container 5 for the wire mesh carrier includes a first holding container 502. The first holding container 502 has an annular groove 509. The annular groove 509 has a wire mesh carrier placement rack 503. The wire mesh carrier placement rack 503 has multiple wire mesh carrier positioning grooves 504 along its circumference. Each wire mesh carrier positioning groove 504 has a wire mesh carrier 505. The wire mesh carrier 505 has multiple wire mesh slots 506 along its circumference. The wire mesh slots 506 are used to place the sample wire mesh 501. The first holding container 502 has a partition groove 507 in the center. The partition groove 507 has a second holding container 508.

[0039] Additional notches are provided on both sides of the carrier slot 506 to facilitate the insertion and clamping of the tweezer arms.

[0040] The annular trough 509 is used to hold liquid nitrogen, and the second container 508 is used to hold liquid ethane; the two containers have slightly different temperatures.

[0041] The sampling volume of the quantitative pipette 11 is manually set in advance, and the sample is drawn from the sample tube. Then the quantitative pipette 11 is inserted into the fixing clip 1002 for later use.

[0042] Since the sample carrier mesh 501 usually needs to be pre-cooled in liquid nitrogen, the entire cryogenic container 5 of the carrier mesh is positioned on the rotating flange 301. The servo motor of the first linear module 2 drives the first sliding plate 201 to move laterally, and the lower extension rod assembly 702 and the rotating flange 301 rotate, so that the carrier mesh slot 506 moves to be directly below the lower extension rod assembly 702, and the clamping direction of the sample tweezers 9 is correct. The servo motor of the third linear module 7 drives the third sliding plate 701 and the lower extension rod assembly 702 to descend, and the sample tweezers 9 picks up the sample carrier mesh 501 and lifts it to a certain height. The fourth linear module 10 on the side drives the tip of the quantitative pipette 11 to approach the vicinity of the mesh surface of the sample carrier mesh 501, and the linear top cylinder 12 pushes the button of the quantitative pipette 11 to add the sample to the mesh surface of the sample carrier mesh 501.

[0043] The lower extension rod assembly 702 moves upward so that the sample carrier mesh 501 is at the liquid suction clamp 8. The liquid suction clamp 8 clamps and contacts the mesh surface of the sample carrier mesh 501 to absorb excess liquid.

[0044] The first sliding plate 201 moves, rotating the flange 301 and the lower extension rod assembly 702, so that the lower end of the sample tweezers 9 is aligned with the second container 508 in the center of the low-temperature container 5. The lower extension rod assembly 702 drives the sample tweezers 9 to move down again into the ethane container of the low-temperature container 5, causing the sample on the sample grid 501 to freeze into a thin layer of ice. The lower extension rod assembly 702 moves up, the first sliding plate 201 moves, rotating the flange 301 and the lower extension rod assembly 702, so that the lower end of the sample tweezers 9 is aligned with the original storage position of the sample grid 501, and the sample grid 501 is put back. The first sliding plate 201 moves, rotating the flange 301 and the lower extension rod assembly 702, so that the sample tweezers 9 is aligned with the storage position of another sample grid 501 in the low-temperature container 5, and then the next round of sample preparation operation is performed.

[0045] Because the quantitative liquid dosing gun technology is relatively mature and the quantitative liquid aspiration operation is also quite convenient, the workload and intensity are not high. Apart from this point, the rest of the complex sample preparation process is automated, avoiding the uncertainty of manual operation and enhancing the sample preparation success rate and the reliability of test detection.

[0046] In a preferred embodiment, the lower extension rod assembly 702 includes a core rod 703, the lower end of which is provided with a detachable connecting rod 719. The sample tweezers 9 are connected to the lower end of the connecting rod 719. The third sliding plate 701 is provided with a rotatable rotating seat 709, and the rotating seat 709 is provided with an end fixing seat 710. The upper end of the core rod 703 is sleeved with the end fixing seat 710.

[0047] In a preferred embodiment, the lower extension rod assembly 702 further includes an outer sleeve 704 sleeved on the outside of the core rod 703. The outer sleeve 704 is threadedly connected to the core rod 703. A rotatable guide sleeve 705 is also provided below the rotating seat 709 on the third sliding plate 701. A straight guide groove 708 is provided along the circumferential direction on the inner wall of the guide sleeve 705. A guide strip 707 is provided on the outer arm of the outer sleeve 704. The guide strip 707 is slidably engaged with the straight guide groove 708. A locking sleeve 711 is provided at the lower end of the outer sleeve 704. The lower end of the locking sleeve 711 presses the arms of the sample forceps 9 to clamp the arms.

[0048] The third sliding plate 701 has a horizontal plate on its side for mounting the bearing seat 706, and the guide sleeve 705 is inserted into the bearing seat 706.

[0049] The third sliding plate 701 is also equipped with a first drive motor 723 and a second drive motor 724. The first drive motor 723 drives the rotating seat 709 to rotate through a synchronous belt mechanism, and the second drive motor 724 drives the guide sleeve 705 to rotate through a synchronous belt mechanism.

[0050] In a preferred embodiment, the upper end of the locking sleeve 711 is provided with a sliding part 713, and the outer wall of the sliding part 713 is provided with an annular locking groove 714. The lower end of the outer sleeve 704 is also provided with a connecting sleeve 715. The lower end of the connecting sleeve 715 has a plurality of deformable arms 716 along the circumference. The inner side of the deformable arm 716 is provided with a locking protrusion 717. The locking protrusion 717 slides and engages with the annular locking groove 714 so that the locking sleeve 711 can rotate relative to the connecting sleeve 715. The lower end of the locking sleeve 711 is provided with a flexible bushing 712. The outer side of the forceps arm of the sample loading forceps 9 is provided with a protrusion 901. The flexible bushing 712 is engaged at the protrusion 901. The outer side of the deformable arm 716 is covered with a limiting sleeve 718.

[0051] The core rod 703 can rotate to adjust the opening direction of the sample loading forceps 9.

[0052] The rotation of the guide sleeve 705 drives the outer sleeve 704 to rotate. Since the upper end of the outer sleeve 704 is threadedly connected to the core rod 703, the outer sleeve 704 rotates and moves down until the locking sleeve 711 contacts the protrusion 901, squeezing the tweezers arm together. The deformation of the flexible bushing 712 enhances the wrapping effect and increases the friction, so that the locking sleeve 711 can stably keep the tweezers arm in the closed state.

[0053] The limiting sleeve 718 is fitted on the outer side of the lower end of the connecting sleeve 715 to limit the deformation of the deformable arm 716.

[0054] When manual operation is required, first remove the connecting rod 719 and the sample forceps 9. The limiting sleeve 718 can be moved upward so that the deformable arm 716 can regain its deformability. At this time, pull the locking sleeve 711 downward forcefully. The deformable arm 716 deforms and the locking protrusion 717 disengages from the annular locking groove 714. After the locking sleeve 711 is removed, it can continue to be put on the sample forceps 9 for manual use.

[0055] In a preferred embodiment, the liquid suction clamp 8 includes two clamping segments 801 arranged opposite to each other. A filter paper sheet 811 is provided on one side of the clamping segment 801, and a connecting arm 802 is provided on the other side of the clamping segment 801. An inner bend 803 is provided at the upper end of the connecting arm 802. A hinge seat 807 and a connecting seat 808 are provided on the base frame. The middle part of the connecting arm 802 is hinged to the hinge seat 807. A tension spring 809 is connected to the connecting seat 808. A connecting ear 810 is provided at the lower end of the connecting arm 802. The tension spring 809 pulls the connecting ear 810. A lifting rod 804 is connected to the lower end of the third sliding plate 701. A fork-shaped block 805 is provided at the lower end of the lifting rod 804. A wedge-shaped surface 806 is provided on the inner side of the fork-shaped block 805. The wedge-shaped surface 806 presses the inner bend 803 to make the clamping segment 801 swing.

[0056] The lower end of the third sliding plate 701 is provided with two sets of lifting rods 804 and related structures. When the third sliding plate 701 is raised, the two clamping petals 801 can swing inward and the lower ends of the clamping petals 801 close. At the same time, the lower extension rod assembly 702, along with the sample tweezers 9, moves to this position. The filter paper 811 on the inner side of the clamping petals 801 contacts the sample carrier mesh 501 and absorbs excess liquid.

[0057] When the lower extension rod assembly 702 moves downward, the clamping flap 801 opens and resets under the tension of the tension spring 809.

[0058] Because the sample carrier 501 is small and the filter paper 811 is large, when the clips 801 are clamped together once, the sample carrier 501 rests against a small area near the bottom of the filter paper 811. To avoid cross-contamination of the sample during the next use, the filter paper was previously replaced directly during manual operation, resulting in low filter paper utilization and waste.

[0059] In a preferred embodiment, the clamping flap 801 includes an end cap 814 and a rear housing 815. The rear housing 815 is connected to the connecting arm 802. A reciprocating movable sleeve 817 is provided inside the rear housing 815. One end of the movable sleeve 817 has a front ring tooth 818, and the other end has a rear ring tooth 821. The end cap 814 has a front ring tooth 816 near the movable sleeve 817, and the front ring tooth 818 meshes with the front ring tooth 816. A rear fixing seat 822 is provided on the base, and a rear ring tooth is provided at the end of the rear fixing seat 822 near the rear housing 815. 823, the rear ring tooth 821 meshes with the rear end ring tooth 823, and a rotating disk 813 is also provided. One end of the rotating disk 813 passes through the end cover body 814 to connect with the movable sleeve 817. The end face of the rotating disk 813 is provided with a deformable flexible ring 812. The filter paper 811 is provided on the flexible ring 812. The front end ring tooth 816, the front ring tooth 818, the rear ring tooth 821 and the rear end ring tooth 823 are inclined teeth. The meshing of the rear ring tooth 821 with the rear end ring tooth 823 and the meshing of the front ring tooth 818 with the front end ring tooth 816 both cause the rotating disk 813 to rotate circumferentially in the same direction.

[0060] The flexible ring 812 can be made of sponge sheet. When the lower edges of the clamping segments 801 are clamped together, the flexible ring 812 can adapt to deformation, ensuring that the filter paper sheet 811 and the sample carrier 501 have full contact without dead corners.

[0061] Because of the oblique teeth, the movable sleeve 817 rotates circumferentially by an angle each time it contacts the teeth at both ends, causing the end cap 814 to rotate by the same angle. This means the area of ​​the filter paper 811 that was wetted by excess liquid from the sample grid 501 rotates to the next circumferential position. Therefore, the next time the clips 801 are clamped, the dry area will contact the sample grid 501. Furthermore, multiple sample preparations can be completed by attaching the filter paper 811 once, resulting in high utilization of the filter paper 811 and eliminating the need for frequent manual replacement, thus increasing the level of automation.

[0062] In a preferred embodiment, a magnetic ring 819 is provided on one side of the movable sleeve 817, and multiple electromagnets 820 are provided circumferentially on the rear housing 815. When the electromagnets 820 are energized, they generate a repulsive force on the magnetic ring 819, which can push the movable sleeve 817 close to the end cover 814 so that the front ring tooth 818 engages with the front end ring tooth 816. When the electromagnets 820 are de-energized, the movable sleeve 817 is reset under the tension of the tension spring 809 so that the rear ring tooth 821 engages with the rear end ring tooth 823.

[0063] Since the swing amplitude of the connecting arm 802 is not large, the electromagnet 820 can always overcome the tension of the tension spring 809 when it is energized, so that the movable sleeve 817 can be pushed forward into place.

[0064] After the clamping of the clamping plates 801 ends and the connecting arm 802 returns to its original position, when the rear ring tooth 821 approaches the rear ring tooth 823, the electromagnet 820 is de-energized again to avoid idle travel and uncertainty in tooth engagement.

[0065] In a preferred embodiment, the end fixing seat 710 is provided with a rotatable ball joint sleeve 720 which is threadedly connected to the core rod 703. The side wall of the end fixing seat 710 is provided with a plurality of threaded tightening screws 722 along the circumferential direction, and one end of each tightening screw 722 abuts against the outer wall of the core rod 703.

[0066] The length of the extension of the core rod 703 can be adjusted by rotation. At the same time, the tightening screws 722 are adjusted to make the core rod 703 swing to adjust its verticality. Then, the tightening screws 722 are tightened and the locking nut 721 is installed to lock the core rod 703.

[0067] In the preferred embodiment, the lower end of the outer sleeve 704 is provided with a lower bushing 725, which is sleeved with the core rod 703.

[0068] The lower bushing 725 can be made of ceramic or coated with Teflon to reduce sliding friction.

[0069] The above embodiments are merely preferred technical solutions of the present invention and should not be considered as limitations on the present invention. The scope of protection of the present invention should be limited to the technical solutions described in the claims, including equivalent substitutions of the technical features described in the claims. That is, equivalent substitutions and improvements within this scope are also within the scope of protection of the present invention.

Claims

1. An automated cryo-electron microscopy sample preparation enhancement device, characterized in that: The system includes a first linear module (2) arranged horizontally, which has a movable first sliding plate (201). A turntable device (3) is mounted on the first sliding plate (201), and a rotatable rotating flange (301) is mounted on the rotating flange (301). A low-temperature container (5) carrying a wire mesh is mounted on the rotating flange (301). A third linear module (7) is arranged vertically above the first linear module (2). The third linear module (7) has a movable third sliding plate (701), and a rotatable lower extension rod assembly (70) is mounted on the third sliding plate (701). 2) The lower end of the lower extension rod assembly (702) is provided with sample tweezers (9) and liquid suction clip (8). The upper end of the liquid suction clip (8) is connected to the third linear module (7). The third linear module (7) is provided with a horizontally arranged fourth linear module (10) on one side. The fourth linear module (10) is provided with a movable fourth sliding plate (1001). The fourth sliding plate (1001) is provided with a fixing clip (1002). The fixing clip (1002) is provided with a quantitative pipette (11). The rear end of the quantitative pipette (11) on the fourth sliding plate (1001) is provided with a linear top cylinder (12).

2. The automated cryo-electron microscopy sample preparation enhancement device according to claim 1, characterized in that: [The device is equipped with a] carrier... The low-temperature container (5) includes a first holding container (502), an annular groove (509) is provided in the first holding container (502), a grid holder (503) is provided in the annular groove (509), a grid holder positioning groove (503) is provided in the grid holder (503) along the circumference, a grid holder (504) is provided in each grid holder positioning groove (504), a grid holder (505) is provided in each grid holder (505) along the circumference, a grid holder slot (506) is provided in the grid holder slot (506) for placing the sample grid (501), a partition groove (507) is provided in the center of the first holding container (502), and a second holding container (508) is provided in the partition groove (507).

3. The automated cryo-electron microscopy sample preparation enhancement device according to claim 1, characterized in that: The lower extension rod assembly (702) includes a core rod (703), the lower end of which is provided with a detachable connecting rod (719). The sample tweezers (9) are connected to the lower end of the connecting rod (719). A rotatable rotating seat (709) is provided on the third sliding plate (701), and an end fixing seat (710) is provided on the rotating seat (709). The upper end of the core rod (703) is sleeved with the end fixing seat (710).

4. The automated cryo-electron microscopy sample preparation enhancement device according to claim 3, characterized in that: The lower extension rod assembly (702) also includes an outer sleeve (704) sleeved on the outside of the core rod (703). The outer sleeve (704) is threadedly connected to the core rod (703). A rotatable guide sleeve (705) is also provided below the rotating seat (709) on the third sliding plate (701). A straight guide groove (708) is provided along the circumferential direction on the inner wall of the guide sleeve (705). A guide strip (707) is provided on the outer arm of the outer sleeve (704). The guide strip (707) is slidably engaged with the straight guide groove (708). A locking sleeve (711) is provided at the lower end of the outer sleeve (704). The lower end of the locking sleeve (711) presses the arms of the sample forceps (9) to make the arms clamp together.

5. The automated cryo-electron microscopy sample preparation enhancement device according to claim 4, characterized in that: The upper end of the locking sleeve (711) is provided with a sliding part (713), and the outer wall of the sliding part (713) is provided with an annular snap-fit ​​groove (714). The lower end of the outer sleeve (704) is also provided with a connecting sleeve (715). The lower end of the connecting sleeve (715) is provided with multiple deformable arms (716) along the circumference. The inner side of the deformable arm (716) is provided with a snap-fit ​​protrusion (717). The snap-fit ​​protrusion (717) and the annular snap-fit ​​groove (714) are slidably snapped together so that the locking sleeve (711) can rotate relative to the connecting sleeve (715). The lower end of the locking sleeve (711) is provided with a flexible bushing (712). The outer side of the forceps arm of the sample loading forceps (9) is provided with a protrusion (901). The flexible bushing (712) is snapped at the protrusion (901). The outer side of the deformable arm (716) is covered with a limiting sleeve (718).

6. The automated cryo-electron microscopy sample preparation enhancement device according to claim 1, characterized in that: The liquid suction clamp (8) includes two clamping flaps (801) arranged opposite to each other. A filter paper sheet (811) is provided on one side of the clamping flap (801), and a connecting arm (802) is provided on the other side of the clamping flap (801). An inner bend (803) is provided at the upper end of the connecting arm (802). A hinge seat (807) and a connecting seat (808) are provided on the base frame. The middle part of the connecting arm (802) is hinged to the hinge seat (807). A tension spring (809) is connected at the connecting seat (808). A connecting ear (810) is provided at the lower end of the connecting arm (802). The tension spring (809) pulls the connecting ear (810). A lifting rod (804) is connected at the lower end of the third sliding plate (701). A fork-shaped block (805) is provided at the lower end of the lifting rod (804). A wedge-shaped surface (806) is provided on the inner side of the fork-shaped block (805). The wedge-shaped surface (806) squeezes the inner bend (803) to make the clamping flap (801) swing.

7. The automated cryo-electron microscopy sample preparation enhancement device according to claim 6, characterized in that: clamping... The flap (801) includes an end cap (814) and a rear housing (815). The rear housing (815) is connected to the connecting arm (802). The rear housing (815) is provided with a movable sleeve (817) that can reciprocate. One end of the movable sleeve (817) is provided with a front ring tooth (818), and the other end of the movable sleeve (817) is provided with a rear ring tooth (821). The end cap (814) is provided with a front ring tooth (816) at the end near the movable sleeve (817). The front ring tooth (818) meshes with the front ring tooth (816). The base is provided with a rear fixing seat (822), and the rear fixing seat (822) is provided with a rear ring tooth (823) at the end near the rear housing (815). The rear ring tooth (821) meshes with the rear ring tooth (823), and a rotating disk (813) is also provided. One end of the rotating disk (813) passes through the end cover body (814) to connect with the movable sleeve (817). The end face of the rotating disk (813) is provided with a deformable flexible ring (812). The filter paper (811) is provided on the flexible ring (812). The front ring tooth (816), the front ring tooth (818), the rear ring tooth (821) and the rear ring tooth (823) are inclined teeth. The meshing of the rear ring tooth (821) with the rear ring tooth (823) and the meshing of the front ring tooth (818) with the front ring tooth (816) both cause the rotating disk (813) to rotate circumferentially in the same direction.

8. The automated cryo-electron microscopy sample preparation enhancement device according to claim 7, characterized in that: A magnetic ring (819) is provided on one side of the movable sleeve (817), and multiple electromagnets (820) are provided along the circumferential direction on the rear housing (815). When the electromagnets (820) are energized, they generate a repulsive force on the magnetic ring (819), which can push the movable sleeve (817) close to the end cover (814) so ​​that the front ring teeth (818) mesh with the front end ring teeth (816). When the electromagnets (820) are de-energized, the movable sleeve (817) is reset under the pulling force of the tension spring (809) so that the rear ring teeth (821) mesh with the rear end ring teeth (823).

9. The automated cryo-electron microscopy sample preparation enhancement device according to claim 3, characterized in that: The end fixing seat (710) is provided with a rotatable ball joint sleeve (720) which is threadedly connected to the core rod (703). The side wall of the end fixing seat (710) is provided with multiple threaded tightening screws (722) along the circumferential direction, and one end of each tightening screw (722) abuts against the outer wall of the core rod (703).

10. The automated cryo-electron microscopy sample preparation enhancement device according to claim 4, characterized in that: The lower end of the outer sleeve (704) is provided with a lower bushing (725), which is sleeved with the core rod (703).

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