A cryo-em grid
By coating the surface of the carrier mesh with a titanium dioxide nano-coating and designing a detachable dual carrier mesh structure, the problems of carrier mesh impurity adsorption and sample solution cross-flow were solved, thereby improving the imaging quality and detection accuracy of cryo-electron microscopy.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- LUOJIADA ADVANCED TECH RES INST OF SUZHOU IND PARK
- Filing Date
- 2025-05-29
- Publication Date
- 2026-06-16
AI Technical Summary
Impurities are easily adsorbed on the surface of the screen, which leads to a decrease in the imaging quality of the electron microscope. Furthermore, when multiple screens are assembled, the sample solution is prone to cross-flow, affecting the detection accuracy and reliability.
A titanium dioxide nano-coating is used to coat the surface of the grid to reduce impurity adsorption, and a detachable dual-grid structure ensures sample independence. The grid spacing is stabilized by using columns and locking components.
It reduces background noise, improves imaging clarity, prevents sample solution cross-flow, and enhances detection accuracy and reliability, making it suitable for high-precision cryo-electron microscopy.
Smart Images
Figure CN224366828U_ABST
Abstract
Description
Technical Field
[0001] This utility model relates to the field of cryo-electron microscopy, specifically to a cryo-electron microscopy support grid. Background Technology
[0002] In the field of cryo-electron microscopy, the application of grids faces numerous challenges. On the one hand, the surface of the grid readily adsorbs various impurities, which interfere with the interaction between the electron beam and the sample, leading to a significant increase in background noise during electron microscopy imaging. This results in reduced image clarity and difficulty in improving resolution, failing to meet the growing demand for high-precision imaging in current scientific research and severely hindering the in-depth advancement of related research.
[0003] On the other hand, existing technologies often employ dual-grid mounting or assembling multiple grids together to simultaneously load multiple samples, aiming to improve work efficiency. However, in practice, when multiple grids are tightly assembled for sample loading, the small spacing between the grids easily leads to cross-flow of different sample solutions. This affects the integrity of each sample, causing deviations in the detection results and significantly reducing detection accuracy, thus greatly impacting the effectiveness and reliability of cryo-electron microscopy in high-precision detection.
[0004] In view of this, it is necessary to provide an improved technical solution to solve the above problems. Utility Model Content
[0005] The purpose of this invention is to provide a cryo-electron microscope grid to address the aforementioned shortcomings in the prior art.
[0006] To achieve the above objectives, this utility model provides the following technical solution:
[0007] A cryo-electron microscope grid, comprising:
[0008] At least two carrier nets, namely a first carrier net and a second carrier net, and both surfaces are provided with a protective coating;
[0009] The first carrier net is provided with a column, and the second carrier net is detachably mounted on the column and can be horizontally rotated or directly engaged relative to the first carrier net for positioning. The edges of the first carrier net and the second carrier net are separated from each other.
[0010] As a preferred embodiment of the present invention, both the first and second carrier nets include a base with holes and a porous silicon nitride film disposed on the base.
[0011] As a preferred embodiment of this utility model, the protective coating is a titanium dioxide nano-coating and is disposed on the porous silicon nitride film.
[0012] As a preferred embodiment of this utility model, a first support part and a hollow support bottom connected horizontally to the edge are vertically arranged on the base of the first carrier net. The base, the first support part and the support bottom constitute a frame structure and the three form an assembly space.
[0013] As a preferred embodiment of the present invention, a first support portion is vertically provided on the base of the first carrier net near the edge, the base and the first support portion forming a semi-open structure, and the two forming an assembly space.
[0014] In a preferred embodiment of this invention, one end of the column is higher than the surface of the first netting.
[0015] As a preferred embodiment of this utility model, the outer surface of the column facing the first carrier net is provided with short positioning protrusions or long positioning protrusions.
[0016] As a preferred embodiment of this utility model, a second support portion is vertically provided on the base of the second carrier net near the edge;
[0017] The second carrier net has a locking part in the middle position. The locking part is composed of symmetrically arranged clamping ends, and the two clamping ends form a notch for assembling and locking the column.
[0018] As a preferred embodiment of this utility model, the engaging part is provided with a positioning groove that cooperates with the short positioning protrusion or the long positioning protrusion.
[0019] As a preferred embodiment of this utility model, a handle is provided at one edge end of the second carrier net for pulling the second carrier net to rotate.
[0020] This invention offers the following advantages: A self-cleaning titanium dioxide nano-coating is applied to the surface of the carrier mesh, utilizing the unique properties of nanomaterials to reduce impurity adsorption and automatically remove impurities, thereby lowering background noise. Furthermore, the multi-carrier mesh configuration allows for simultaneous testing of multiple samples, and its assembly structure ensures the rationality and stability of the mesh spacing, effectively preventing cross-contamination of sample solutions. Attached Figure Description
[0021] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in this utility model. For those skilled in the art, other drawings can be obtained based on these drawings.
[0022] Figure 1This is a schematic diagram of the first and second carrier nets of this utility model in their assembly and use state.
[0023] Figure 2 This is a schematic diagram of another assembly structure of the first and second carrier nets of this utility model.
[0024] Figure 3 This is a schematic diagram of the first carrier network structure of this utility model.
[0025] Figure 4 This is a schematic diagram of the second carrier net structure of this utility model.
[0026] Figure 5 This is a schematic diagram of the base, porous silicon nitride film, and protective coating structure of the two-carrying mesh of this utility model.
[0027] Figure 6 This is a schematic diagram of the structure of the first carrier net in another embodiment of this utility model.
[0028] Figure 7 This is a schematic diagram of the assembly structure of the first and second carrier nets according to another embodiment of the present invention.
[0029] Figure 8 This is a schematic diagram of the structure of the long positioning protrusion on the first carrier wire in another embodiment of the present invention.
[0030] Explanation of reference numerals in the attached figures:
[0031] 10. Protective coating; 20. Porous silicon nitride film; 30. Base; 100. First carrier net; 11. Column; 12. First support part; 13. Support bottom; 14. Assembly space; 15. Short positioning protrusion; 16. Long positioning protrusion; 200. Second carrier net; 21. Second support part; 22. Engaging part; 221. Clamping end; 222. Positioning groove; 23. Handle. Detailed Implementation
[0032] The technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments. Obviously, the described embodiments are only some embodiments of the present utility model, and not all embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the protection scope of the present utility model.
[0033] This invention provides a cryo-electron microscope (CEMM) support, which is a flat disc with a grid structure or other shaped holes. Its main function is to support and fix the sample during transmission electron microscopy (TEM) observation, ensuring the sample remains stable during TEM observation. The use of the support and sample loading for observation are existing technologies and will not be elaborated upon.
[0034] Specifically, such as Figures 1 to 5 As shown, it includes at least two carrier nets, namely a first carrier net 100 and a second carrier net 200, and both surfaces are provided with a protective coating 10.
[0035] Among them, such as Figure 5 As shown, both the first carrier mesh 100 and the second carrier mesh 200 include a base 30 with perforations and a porous silicon nitride film 20 disposed on the base 30. The base 30 uses silicon as the main material, possessing excellent mechanical strength and chemical inertness, maintaining structural stability under extreme low temperatures (liquid nitrogen temperature) and high vacuum environments, avoiding the risk of deformation or breakage. Simultaneously, it facilitates precision etching and micro / nano fabrication, allowing for the customization of carrier mesh structures with different pore sizes and arrangements. The porous silicon nitride film 20 is disposed on the surface of the base 30, forming a unified whole with the base 30. Using a silicon nitride film ensures efficient electron beam penetration, thereby obtaining high-quality imaging results. It also avoids carbon buildup and possesses characteristics such as acid resistance, high temperature resistance, electron beam irradiation resistance, uniform imaging background, and low noise. It is also suitable for difficult-to-process samples such as membrane proteins and liposomes; the hydrophilicity of the silicon nitride film can optimize the quality of thin ice layer formation.
[0036] Furthermore, the protective coating 10 employs a titanium dioxide nanocoating and is disposed on the porous silicon nitride film 20. Alternatively, a zinc oxide nanocoating or a carbon nanocoating can also be used. Preferably, titanium dioxide generates reactive oxygen species (ROS) under ultraviolet light (or electron beam excitation), which can decompose adsorbed organic pollutants (such as protein residues, lipids, etc.), giving the support network self-cleaning ability and reducing background noise during sample preparation.
[0037] Titanium dioxide nanostructures can achieve superhydrophobicity (contact angle >150°) or selective hydrophilicity by calcining to regulate surface energy, thereby reducing the random spread of sample solution on the carrier surface and making the thin ice layer more uniformly distributed.
[0038] Titanium dioxide nanocoatings exhibit excellent chemical stability and strong chemical inertness, enabling them to withstand acidic environments (such as pH 2-11) and high-energy electron beams (200-300 keV) during electron microscopy sample preparation, thus preventing degradation of the film material.
[0039] The preparation process of the nano-coating on the carrier mesh is as follows: a titanium dioxide nano-coating is prepared on the carrier mesh surface using a sol-gel method. First, tetrabutyl titanate, anhydrous ethanol, deionized water, and hydrochloric acid are mixed in a certain proportion and stirred evenly to form a sol. Then, the carrier mesh is immersed in the sol, and the sol is uniformly coated onto the carrier mesh surface using a dip-coating method. Finally, the carrier mesh coated with sol is calcined at high temperature to transform the sol into a titanium dioxide nano-coating. The thickness of the coating can be adjusted by controlling the concentration of the sol and the dip-coating speed, and is generally 50-200 nm thick. Therefore, the titanium dioxide nano-coating on the carrier mesh simultaneously possesses good liquid affinity and anti-fouling properties, as well as stability and durability, showing significant effectiveness in practical applications.
[0040] In some embodiments, a column 11 is provided on the first net 100, and a second net 200 is detachably disposed on the column 11 and can be horizontally rotated or directly engaged relative to the first net 100 for positioning. The edges of the first net 100 and the second net 200 are spaced apart from each other.
[0041] After the second mesh 200 is assembled with the first mesh 100, it can be pulled to rotate horizontally around the cylinder 11 as needed for the experiment, precisely separating the edges of the two meshes (the spacing is selected according to the experimental requirements), forming a physical isolation barrier, which greatly prevents sample solution cross-contamination, especially suitable for easily diffusing liquid samples. Once the second mesh 200 is rotated into position, it engages and is fixed to the first mesh 100, ensuring a constant spacing during freezing, transfer, and electron microscopy observation, preventing displacement deviations caused by vibration or operation. Figure 2 As shown in the attached diagram, this only illustrates the assembly of the second carrier net 200 and the first carrier net 100, not their operational state. In the operational state, there are no obstructions below the first carrier net 100 and the second carrier net 200. Figure 1 As shown.
[0042] Furthermore, the dual-grid setup allows for the simultaneous loading of control and experimental group samples, or the same sample at different concentrations / conditions, reducing batch errors and improving data comparability. On the other hand, the observation area can be quickly switched by rotating the grid in the electron microscope without changing the sample stage, saving equipment usage time. Additionally, when assembling the first grid 100 and the second grid 200, the second grid 200 can be directly snapped onto the column 11 without requiring rotation and repositioning before snapping. These two configurations can be selected according to experimental needs.
[0043] To better understand this utility model, some specific embodiments of the first carrier net 100 and the second carrier net 200 will be further described in detail:
[0044] In some embodiments, a first support portion 12 and a horizontally connected perforated support bottom 13 are vertically arranged near the edge of the base 30 of the first mesh 100. The base 30, the first support portion 12, and the support bottom 13 constitute a frame structure, and the three form an assembly space 14. This frame structure obviously enhances the overall stability of the mesh. It can also effectively disperse and withstand external forces, reduce stress concentration caused by sample weight or operation, thereby reducing the risk of mesh deformation and ensuring stability during use. The assembly space 14 improves the ease of assembly of the first mesh 100 and the second mesh 200, reducing unnecessary contact between them, allowing for plug-and-play operation. The perforated support bottom 13 is used because the surface of the mesh has holes, and the electron beam of the transmission electron microscope needs to pass through the holes to observe the liquid sample. The absence of any obstruction below the holes results in clearer imaging.
[0045] In other embodiments, such as Figure 6 and 7 As shown, a first support portion 12 is vertically arranged near the edge of the base 30 of the first carrier 100. The base 30 and the first support portion 12 form a semi-open structure, and the two form an assembly space 14. The plate-open structure provides an open loading space, which makes the two carriers simpler, and the manufacturing complexity and cost of the first carrier 100 are greatly reduced. At the same time, the simplified structure reduces the interference encountered by the electron beam when passing through the sample.
[0046] Furthermore, one end of the column 11 is higher than the surface of the first mesh 100. By setting the column 11 above the surface, a physical barrier is formed, which can effectively prevent the sample liquid from splashing out or flowing onto the adjacent second mesh 200 when dripping. This greatly reduces mixing and interference between samples, ensuring the independence and integrity of each sample.
[0047] like Figure 3 and 8 As shown, the column 11 has short positioning protrusions 15 or long positioning protrusions 16 on its outer surface facing the first carrier net 100. A second support portion 21 is vertically arranged near the edge of the base 30 of the second carrier net 200; a locking portion 22 is provided in the middle of the second carrier net 200, which is composed of symmetrically arranged clamping ends 221, each clamping end 221 forming a notch for assembling and locking the column 11. The locking portion 22 has positioning grooves 222 that mate with the short positioning protrusions 15 or long positioning protrusions 16.
[0048] When the second net 200 is assembled, it first abuts against the column 11 through the two clamping ends 221 at the notch. As the second net 200 is continuously squeezed, the two clamping ends 221 cooperate with the column 11. A handle 23 is provided at one edge of the second net 200. At this time, when the handle 23 is pulled, the second net 200 rotates, and the rotating positioning groove 222 cooperates with the short positioning protrusion 15, and the installation of the second net 200 is completed.
[0049] In other embodiments, such as Figure 8 As shown, the difference between the second carrier net 200 in the above embodiment and the assembly method is that when the two clamping ends 221 are engaged with the column 11, the long positioning protrusion 16 extends into the groove 222. The depth of the groove 222 can accommodate one end of the long positioning protrusion 16, so that the second carrier net 200 can be directly engaged and fixed with the first carrier net 100, which simplifies the operation.
[0050] In another embodiment, a micro / nano structure for mounting a MEMS chip is disposed within the base 30 of the first carrier mesh 100 or the second carrier mesh 200, and the chip is fixed by micro-welding, bonding, or other micromechanical structures. The MEMS chip includes components such as micropumps, microvalves, and sensors. Microfluidic channels are designed on the carrier mesh or chip, and the sample liquid is guided to flow on the carrier mesh by precisely controlling the shape and size of the channels. The micropump provides the power for the liquid flow, and the flow rate of the liquid can be precisely adjusted by controlling the operating frequency and voltage of the micropump. The microvalves control the direction and flow rate of the liquid, achieving precise liquid distribution. The sensors monitor parameters such as the liquid flow rate, pressure, and temperature in real time, and feed them back to the control system so that the operating status of the micropumps and microvalves can be adjusted in a timely manner. Through the microfluidic channel structure and precise control of the MEMS chip, it is possible to ensure that the liquid is uniformly and stably distributed on the surface of the carrier mesh, eliminating flow around the surface.
[0051] The above description of the disclosed embodiments enables those skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims
1. A cryo-electron microscope grid, characterized in that, include: At least two carrier nets, namely a first carrier net and a second carrier net, and both surfaces are provided with a protective coating; The first carrier net is provided with a column, and the second carrier net is detachably mounted on the column and can be horizontally rotated or directly engaged relative to the first carrier net for positioning. The edges of the first carrier net and the second carrier net are separated from each other.
2. The cryo-electron microscope grid according to claim 1, characterized in that: Both the first and second carrier nets include a base with holes and a porous silicon nitride film disposed on the base.
3. The cryo-electron microscope grid according to claim 2, characterized in that: The protective coating is a titanium dioxide nanocoating and is disposed on the porous silicon nitride film.
4. The cryo-electron microscope grid according to claim 2, characterized in that: The first support part and the hollow support bottom connected horizontally to the base of the first carrier net are vertically arranged near the edge. The base, the first support part and the support bottom form a frame structure and form an assembly space.
5. The cryo-electron microscope grid according to claim 2, characterized in that: A first support portion is vertically arranged on the base of the first carrier net near the edge. The base and the first support portion form a semi-open structure and form an assembly space between them.
6. The cryo-electron microscope grid according to claim 1, characterized in that: One end of the column is higher than the surface of the first net.
7. The cryo-electron microscope grid according to claim 1, characterized in that: The column has short or long positioning protrusions on its outer surface facing the first carrier net.
8. The cryo-electron microscope grid according to claim 7, characterized in that: A second support is vertically provided on the base of the second carrier net near the edge; The second carrier net has a locking part in the middle position. The locking part is composed of symmetrically arranged clamping ends, and the two clamping ends form a notch for assembling and locking the column.
9. The cryo-electron microscope grid according to claim 8, characterized in that: The engaging part is provided with a positioning groove that mates with the short positioning protrusion or the long positioning protrusion.
10. The cryo-electron microscope grid according to claim 1, characterized in that: A handle is provided at one edge end of the second carrier net for pulling the second carrier net to rotate.