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Multifunctional graphene grid and preparation method thereof

A multifunctional, graphene-based technology that can be used in measurement devices, instruments, scientific instruments, etc., to solve problems such as the inability to specifically adjust the molecular orientation of target particles

Active Publication Date: 2022-01-21
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these affinity grids cannot specifically adjust the orientation of target particle molecules and may lead to more serious problems of preferred orientations

Method used

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  • Multifunctional graphene grid and preparation method thereof
  • Multifunctional graphene grid and preparation method thereof
  • Multifunctional graphene grid and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1, preparation and structural characterization of MFG carrier network

[0054] 1) Preparation of graphene grid

[0055] First, the CVD method is used to grow graphene film on copper foil, and then the copper foil is directly and selectively etched into the micro-grid of the electron microscope grid to obtain an ultra-clean and strong graphene grid, which contains an array of holes ( figure 1 (A) and figure 1 (B)).

[0056] During low-temperature sample preparation, the coverage of graphene and the integrity on the grid depend on the pore diameter and the number of graphene layers, and when the pore diameter of bilayer or multilayer graphene is about 30 μm, it can be well maintained Graphene coverage and integrity on the grid. In this embodiment, a grid covered with 1 to 4 layers of graphene is used as a starting material. Suspended graphene is ultra-clean, and when imaged by high-resolution STEM, the carbon atoms on the etched copper holes can be well ide...

Embodiment 2

[0061] Embodiment 2, the biological application performance research of MFG carrying net

[0062] 1. The MFG grid is hydrophilic and resistant to electron beam radiation

[0063] The present invention checks hydrophilicity by water contact angle (WCA) test. The present invention has measured the WCA of graphene carrier network and multifunctional graphene carrier network, finds that after functionalization, this angle reduces to 39.1 ° from 97.4 ° ( figure 2 (A)), suitable for subsequent bioelectron microscopy specimen preparation. The superior properties of graphene, such as ultrahigh thermal and electrical conductivity and strong mechanical strength, are mainly attributed to the perfect two-dimensional hexagonal lattice formed by sp2 hybridized carbon atoms. Raman spectroscopy shows that there is no D peak (1350cm -1 ), indicating that graphene has almost no defects ( figure 2 (B), upper blue curve). After multiple functionalization, a weak D peak appeared, which was ...

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Abstract

The invention discloses a multifunctional graphene grid (MFG grid) and a preparation method thereof. The preparation method of the MFG grid comprises the following steps: etching a copper foil covered by a CVD grown graphene film as an electron microscope grid to obtain a graphene grid; performing oxygen plasma treatment on the graphene grid; then, adopting an MES buffer solution containing EDC and NHS to treat the graphene grid; activating the graphene carrying net by adopting nitrilotriacetic acid and biotin; and finally reacting with nickel salt. The MFG grid can specifically anchor target biomolecules on the surface of the MFG carrier network, so that the interface problem of air and water is avoided, the orientation distribution of particles can be controlled, and abundant Euler angles are provided by constructing related protein purification tags in different directions of each biomacromolecule, and moreover, the structural analysis of biomacromolecules is more reliable and more efficient, so that the MFG grid can promote the universality and repeatability of application of the cryoelectron microscope in structural biological analysis.

Description

technical field [0001] The invention relates to a multifunctional graphene carrier net and a preparation method thereof, belonging to the technical field of biological cryo-electron microscopy. Background technique [0002] Due to the continuous development of cryo-EM technology-related hardware and software, cryo-electron microscopy (cryo-EM) is now a routine method for solving atomic-resolution structures of biological macromolecules, which greatly expands the understanding of biological mechanisms at the molecular level . However, this method still faces some key technical challenges, especially for frozen sample preparation. In practice, a small amount of a biomolecule-containing solution is added dropwise onto a freshly glow-discharged grid covered by a support membrane (eg, a porous carbon membrane) and then quenched in liquid ethane. Ideally, biomolecules would be expected to be embedded in a directionally enriched manner in a thin vitrifiable ice layer supported by...

Claims

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Application Information

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IPC IPC(8): G01N23/2202G01N23/2251
CPCG01N23/2202G01N23/2251G01N2223/07G01N2223/102G01N2223/3103
Inventor 王宏伟陈亚楠刘楠胡翠霞
Owner TSINGHUA UNIV
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