High-throughput repeatable multifunctional film fishing device

Abstract

The invention discloses a high-throughput repeatable multifunctional film fishing device which can quickly prepare a high-quality ultrathin carbon film carrier net, a graphene carrier net, a graphene oxide film carrier net and the like, can meet the requirement of multi-directional film fishing by accurately adjusting the working distance of film fishing and the angle of film fishing, replaces manual film fishing by mechanical film fishing, and has the advantages of high yield, high repeatability and high efficiency. The stability and repeatability of film fishing are improved, and the flux of film fishing is improved by assembling a plurality of multifunctional detachable clamps. The high-flux ultrathin film fishing device is simple and easy to operate, the film fishing process is stable, and a high-integrity ultrathin carbon film is easy to obtain.

Description

TECHNICAL FIELD

[0001] The application belongs to the field of cryo-EM sample preparation, and particularly relates to a high-throughput and repeatable multifunctional membrane fishing device. BACKGROUND

[0002] In the previous transmission electron microscope sample preparation process, a continuous support film is often laid on the support grid to support, stabilize and protect the sample. Especially for cryo-electron microscopy (cryo-EM), sample preparation is one of the main bottleneck problems of cryo-EM technology, and high-throughput and repeatable support grids are urgently needed for cryo-EM sample preparation. As an important biological imaging technology, cryo-EM is used to observe the structure and dynamic process of biomolecules and cells in a natural state, and the requirement of high-resolution structure research is to rapidly freeze the biological sample in a thin layer of glassy ice. However, the support grid used in the conventional cryo-EM sample preparation is mainly made of amorphous carbon film or silicon-nitrogen film, which has poor thermal and electrical conductivity and relatively weak mechanical strength, and can bring strong background noise and serious beam movement to the imaging process. In addition, it can also cause the air-water interface to induce protein denaturation / degradation, the protein particles are not very ideal in the hole, and there are problems such as preferred orientation. In the process of cryo-EM sample preparation, the support film is a very important part, which can make the sample keep the natural state and morphology during freezing, slicing and imaging. At present, researchers have developed a variety of thin film materials, such as ultra-thin amorphous carbon film, single-layer lipid film, graphene film, etc.

[0003] The common method for fishing the support film of the support grid is to float the support film on the liquid surface, and then manually fish the single support grid with tweezers. In this process, due to manual fishing, the quality of the fished membrane is uneven and uncontrollable, and the single fishing method takes a long time and has low efficiency. Therefore, it is necessary to develop a high-throughput and repeatable batch fishing device. SUMMARY

[0004] The application aims to solve the technical problems in the background art, and provides a high-throughput and repeatable multifunctional membrane fishing device, which not only improves the fishing efficiency, but also avoids the errors caused by manual fishing, and improves the stability and controllability of the fishing.

[0005] In order to solve the technical problems, the technical scheme of the application is as follows:

[0006] The utility model provides a high throughput repeatable multifunctional membrane fishing device, including: workbench, still including: lifting mechanism, damping shaft and telescopic mechanism, the lifting mechanism fixedly connected workbench, the lifting mechanism is through damping shaft swing joint telescopic mechanism, the workbench is set up and is installed mobile platform on the slide groove, the slide groove cooperation mobile platform slides and uses, the front end of telescopic mechanism is installed and is provided with fishing tool fixed plate, the fishing tool fixed plate is detachably provided with fishing tool.

[0007] Further, first, the lifting mechanism is used to drive the overall movement of the telescopic mechanism, then, the rotation angle of the overall telescopic mechanism is adjusted through the damping shaft, and finally, the fishing process is realized through the cooperation of the telescopic mechanism and the fishing tool fixed plate.

[0008] Further, the bottom of the lifting mechanism is connected to the workbench through the base flange.

[0009] Further, the lifting mechanism comprises a first lead screw, a first adjusting knob and a first housing.

[0010] The lower end of the first housing is fixedly connected to the base, the first lead screw is installed in the first housing, the first adjusting knob installed on the upper end of the first lead screw is used to control the rotary motion of the first lead screw, which indirectly drives the linear motion of the telescopic mechanism, and the lifting limiting guide rail is formed on the first housing to guide and limit.

[0011] Further, the telescopic mechanism comprises a second lead screw, a second adjusting knob, a second housing and a telescopic piece.

[0012] The second lead screw is installed in the second housing, the telescopic piece is installed on the second lead screw, the second adjusting knob is installed on the upper end of the second housing to control the rotary motion of the second lead screw, which indirectly drives the linear motion of the telescopic piece to realize telescoping, and the telescoping limiting guide rail is formed on the second housing to guide and limit.

[0013] Further, bearings connected with the first lead screw and the second lead screw are installed on the lifting limiting guide rail and the telescoping limiting guide rail, the bearings rotate when there is friction, and the bearings slide forward when there is no friction.

[0014] Further, a plurality of object holding cups are uniformly distributed on the mobile platform.

[0015] Further, the fishing tool adopts a self-locking forceps.

[0016] A high-throughput repeatable multifunctional membrane fishing method, the method is applied to the system in any of the above, the method comprises:

[0017] Step one: in the object cup, add solution, release the required film into the solution. The solution is deionized water, the film is 2nm thick ultra-thin carbon film, and the moving stage is moved to the appropriate distance by moving the plane forward and backward;

[0018] Step two: use the film fishing tool to clamp the film fishing substrate, and fix the film fishing tool to the detachable film fishing tool fixing plate. The film fishing tool uses a self-locking tweezers, and the film fishing substrate is a Cu carrier net.

[0019] Step three: adjust the first adjusting knob to adjust the lifting distance of the Z-axis, adjust the height to a position 10cm above the workbench, adjust the second adjusting knob to adjust the distance of the X-axis, that is, the horizontal substrate to the working distance of the object cup, adjust to a position where the distance between them is 15cm, and adjust the angle between the film fishing substrate and the film solution by adjusting the damping shaft between the lifting mechanism and the telescopic mechanism, adjust the water entry angle to 45°.

[0020] Step four: fine-tune the X-axis distance of the second lead screw, adjust the film fishing substrate to below the floating film liquid level, slowly adjust the damping shaft, gradually adjust the angle to about 0°, and then slowly adjust the Z-axis unit at a speed of 10mm / min, so that the substrate rises to a distance of 6cm from the liquid level, and the substrate can be removed. The film fishing step is completed.

[0021] Compared with the prior art, the advantages of the present application are:

[0022] A high-throughput repeatable multifunctional film fishing device can quickly prepare high-quality ultra-thin carbon film carrier nets, graphene carrier nets, graphene oxide film carrier nets, etc. The working distance and angle of film fishing can be accurately adjusted to meet the requirements of multi-directional film fishing. The stability and repeatability of film fishing are improved by replacing human hands with machinery. The throughput of film fishing is improved by assembling multiple multifunctional detachable clamps. The high-throughput ultra-thin film fishing device is simple, easy to operate, stable in film fishing process, and easy to obtain high-integrity ultra-thin carbon film. BRIEF DESCRIPTION OF DRAWINGS

[0023] Figure 1 The figure is a structural schematic diagram of the high-throughput repeatable multifunctional film fishing device of the present application;

[0024] Figure 2 The figure is a structural schematic diagram of the telescopic mechanism of the present application;

[0025] Figure 3 The figure is a structural schematic diagram of the lifting mechanism of the present application;

[0026] Figure 4 The figure is a partial enlarged view of the damping shaft of the present application.

[0027] Reference signs:

[0028] 1-Workbench; 2-Lifting mechanism; 3-Fishing tool fixing plate; 4-Fishing tool; 5-Moving platform; 6-Loading cup; 7-Base; 8-Damping shaft; 9-Telescopic mechanism; 10-Slide groove; 201-First lead screw; 202-First adjusting knob; 203-First housing; 204-Lifting limit guide rail; 901-Second lead screw; 902-Second adjusting knob; 903-Second housing; 904-Telescopic limit guide rail; 905-Telescopic component. Detailed Implementation

[0029] The specific implementation of the present invention is described below with reference to embodiments:

[0030] It should be noted that the structures, proportions, sizes, etc. shown in this specification are only used to complement the content disclosed in the specification for those skilled in the art to understand and read, and are not intended to limit the conditions under which the present invention can be implemented. Any modifications to the structure, changes in the proportions, or adjustments to the size, without affecting the effects and objectives that the present invention can produce, should still fall within the scope of the technical content disclosed in the present invention.

[0031] Furthermore, the terms such as "upper," "lower," "left," "right," "middle," and "one" used in this specification are merely for clarity of description and are not intended to limit the scope of the invention. Any changes or adjustments to their relative relationships, without substantially altering the technical content, should also be considered within the scope of the invention.

[0032] Example 1:

[0033] This embodiment takes a 2nm thick ultrathin carbon film as an example, and will be further described in detail with reference to the accompanying drawings.

[0034] Figure 1 A schematic diagram of the device of the present invention is shown, including a chute 10, which allows the carrier fixing frame holding the film to move back and forth to adjust the distance as needed.

[0035] The multi-directionally adjustable lifting mechanism 2 is fixed to the worktable 1 by a base holder 7. (See attached...) Figure 3 As shown, the lifting mechanism 2 is equipped with a damping shaft, such as... Figure 4 As shown, it can rotate 180°, start and stop at will, and adjust the appropriate film-catching distance and angle before fixing the first lead screw 201 to improve stability during the film-catching process. Meanwhile, adjustable distance knobs are installed at the ends of the first lead screw 201 and the second lead screw 901, respectively. The first lead screw 201 and the vertical guide rail 204 allow precise adjustment of the lifting distance of the second lead screw 901 by adjusting the knob at the end of the first lead screw 201, thus achieving control in the Z-axis direction. Figure 2As shown, the distance of the front and back extension of the second screw rod 901 can be accurately adjusted by adjusting the knob at the end of the second screw rod 901.

[0036] The detachable multi-energy fishing tool fixing plate 3 of the fishing tool (such as tweezers), which can not only fix the fishing tool 4, but also fix a plurality of self-modifiable fishing tools 4 before fishing.

[0037] The moving carrier 5 is usually required to release the film into a container containing a solution, whether it is fishing or demolding. In order to facilitate operation, the device is designed to meet the flux fishing of the moving carrier 5. The fishing carrier cup 6 used by the device is a circular cup, which is fixed on the cup fixing frame with a groove by screws, which can greatly improve the stability of the cup during fishing.

[0038] The base 7 of the fixed lifting mechanism 2 can better maintain the stability of the lifting mechanism 2 during fishing. The base 7 can firmly fix the lifting mechanism 2 on the bottom plate of the workbench, thereby maintaining the stability of the entire device.

[0039] Example 2:

[0040] The method for fishing and demolding ultra-thin film by using the device of the present application comprises the following steps:

[0041] Step one: add a solution to the carrier cup 6, and release the required film into the solution. The solution is deionized water, and the film is an ultra-thin carbon film with a thickness of 2 nm. The moving carrier 5 is moved forward and backward to a suitable distance;

[0042] Step two: clamp the fishing film substrate with the fishing tool 4, and fix the fishing tool 4 to the detachable fishing tool fixing plate 3. The fishing tool 4 is a self-locking tweezer, and the fishing film substrate is a Cu carrier net.

[0043] Step three: adjust the first adjusting knob 202 to adjust the lifting distance of the Z-axis, and adjust the height to a position 1110 cm above the workbench. Adjust the second adjusting knob 902 to adjust the distance of the X-axis, that is, the working distance of the horizontal substrate to the carrier cup 6, to a position where the distance between them is 15 cm. Adjust the damping shaft 8 between the lifting mechanism 2 and the extension mechanism 9 to adjust the angle between the fishing film substrate and the film solution, and adjust the water entry angle to 45°.

[0044] Step four: fine-tune the distance of the X-axis of the second screw rod 901, adjust the film substrate to below the floating film liquid level, slowly adjust the damping shaft 8, gradually adjust the angle to about 0°, and then slowly adjust the Z-axis unit at a speed of 10 mm / min, so that the substrate rises to a distance of 6 cm from the liquid level, and the substrate can be taken down, and the film fishing step is completed.

[0045] The preferred embodiments of the present application are described in detail above, but the present application is not limited to the above-described embodiments, and various changes can be made within the knowledge of those skilled in the art without departing from the spirit of the present application.

[0046] Many other changes and modifications can be made without departing from the spirit and scope of the application. It is to be understood that the application is not limited to the specific embodiments described herein, but only by the claims that follow.

Claims

1. A high-throughput, repeatable, multi-functional membrane fishing device comprising: The workbench (1) is characterized in that it further comprises a lifting mechanism (2), a damping shaft (8) and a telescopic mechanism (9); the lifting mechanism (2) is fixedly connected to the workbench (1), and the lifting mechanism (2) is movably connected to the telescopic mechanism (9) through the damping shaft (8); a sliding groove (10) is formed in the workbench, a movable platform (5) is arranged on the sliding groove (10), the sliding groove (10) is used in sliding cooperation with the movable platform (5), and a fishing tool fixing plate (3) is arranged at the front end of the telescopic mechanism (9); the fishing tool fixing plate (3) is detachably provided with a fishing tool (4).

2. A high throughput repeatable multifunctional membrane skimming device as claimed in claim 1, wherein, Firstly, the lifting mechanism (2) is used for driving the telescopic mechanism (9) to move as a whole, then the rotation angle of the telescopic mechanism (9) as a whole is adjusted through the damping shaft (8), and finally the fishing process is realized through the telescopic mechanism (9) in cooperation with the fishing tool fixing plate (3).

3. A high throughput repeatable multifunctional membrane skimming device as claimed in claim 1, wherein, The bottom of the lifting mechanism (2) is flange-connected to the workbench (1) through a base (7).

4. A high throughput repeatable multifunctional membrane skimming device as claimed in claim 3, wherein, The lifting mechanism comprises a first lead screw (201), a first adjusting knob (202) and a first shell (203). The lower end of the first shell (203) is fixedly connected to the base (7), the first lead screw (201) is arranged in the first shell (203), the first adjusting knob (202) arranged at the upper end of the first lead screw (201) is used for controlling the rotary motion of the first lead screw (201), and the first lead screw (201) indirectly drives the telescopic mechanism (9) to move linearly; the first shell (203) is provided with a lifting limiting guide rail (204) for guiding and limiting.

5. A high throughput repeatable multifunctional membrane skimming device as claimed in claim 4, wherein, The telescopic mechanism (9) comprises a second lead screw (901), a second adjusting knob (902), a second shell (903) and a telescopic part (905). The second lead screw (901) is arranged in the second shell (903), the telescopic part (905) is arranged on the second lead screw (901), the second adjusting knob (902) is arranged at the upper end of the second shell (903) and is used for controlling the rotary motion of the second lead screw (901), the second lead screw (901) indirectly drives the telescopic part (905) to move linearly, realizes telescopic movement, and the second shell (903) is provided with a telescopic limiting guide rail (904) for guiding and limiting.

6. A high throughput repeatable multifunctional membrane skimming device as claimed in claim 5, wherein, Bearings connected with the first lead screw (201) and the second lead screw (901) are arranged on the lifting limiting guide rail (204) and the telescopic limiting guide rail (904), the bearings rotate when there is friction, and the bearings slide forward when there is no friction.

7. A high throughput repeatable multifunctional membrane skimming device as claimed in claim 1, wherein, A plurality of object holding cups (6) are uniformly distributed on the movable platform (5).

8. A high throughput repeatable multifunctional membrane skimming device as claimed in claim 1, wherein, The fishing tool (4) is a self-locking tweezers.

9. A high throughput repeatable multi-functional membrane fishing method characterized by, The method is applied to the system of any one of claims 1-8, and the method comprises the following steps: In step one, a solution is added to the object holding cup (6), a required film is released into the solution, the solution is deionized water, and the film is a 2nm-thick ultrathin carbon film; the movable platform (5) is moved to a suitable distance through plane forward and backward movement. Step two: hold the membrane substrate with the membrane tool (4) and fix the membrane tool (4) to the detachable membrane tool fixing plate (3), the membrane tool (4) is a self-locking tweezers, and the membrane substrate is a Cu carrier net; Step three: adjust the lifting distance of the Z-axis by adjusting the first adjusting knob (202), and adjust the distance of the X-axis by adjusting the second adjusting knob (902), that is, the working distance of the horizontal substrate to the object cup (6), adjust the distance between the two, and adjust the angle between the membrane substrate and the membrane solution by adjusting the damping shaft (8) between the lifting mechanism (2) and the telescopic mechanism (9), and adjust the water entry angle; Step four: fine-tune the distance of the X-axis of the second lead screw (901), adjust the membrane substrate to be below the floating membrane liquid level, slowly adjust the damping shaft (8), and gradually adjust the angle to about 0°, so that the membrane falls on the substrate, then slowly adjust the Z-axis unit to make the substrate rise, and the substrate can be taken out, and the membrane fishing step is completed.

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