▎ 摘　　要

NOVELTY - An electron microscopy grid (6) comprises a perforated substrate (5), a support film (4) on the perforated substrate, a mixture of linker molecules (1,2) comprising (A) at least a linker molecule (M1) (I) and (B) at least a linker molecule (M2) comprising (a) linker molecule (I) (in which at least AS, L1, L2 or a is different from AS, L1, L2 or a of the linker molecule (I)) or (b) a linker molecule (II). The linker molecules are attached to the support film via the anchoring group. USE - Electron microscopy grid e.g. affinity electron microscopy grid used for immobilizing analyte for determining structure of analyte (all claimed) e.g. protein, nucleic acid, carbohydrate, lipid vesicle, nanoparticle, ligand, drug candidate, and small molecule. Can also be used for epitope mapping and antibody screening. ADVANTAGE - The electron microscopy grid provides excellent angular distribution of the analyte once immobilized on the grid, maximizes the angular distribution of the particle, which in turn increasing the number of different projection angles of the analyte and improving the resolution, reduces the time needed for image acquisitioning, allows easy priming of the analyte, prevents further addition of additives or detergents after the analyte bind to the anchoring groups, speeds up sample preparation with more reliable and reproducible manner, reduce noise in the images as long linkers interfere with the image of the analyte, allows a high signal to noise ratio, and prevents interaction between the analyte and the surface of the grid or support film. As the angular distribution of the analyte on the grids is maximized, no need to tilt the grids in different angles during image acquisitioning, which further reduce the time needed for image acquisitioning. DETAILED DESCRIPTION - An electron microscopy grid (6) comprises a perforated substrate (5), a support film (4) on the perforated substrate, a mixture of linker molecules (1,2) comprising (A) at least a linker molecule (M1) of formula (I) and (B) at least a linker molecule (M2) comprising (a) linker molecule (I) (in which at least AS, L1, L2 or a is different from AS, L1, L2 or a of the linker molecule (I)) or (b) a linker molecule of formula (II). The linker molecules are attached to the support film via the anchoring group. AG = anchoring group for anchoring linker molecule to support film; BU = binding unit for binding to analyte; L1-L3 = linear linker section; (a) = angle between linear linker sections;and AS = angled linker section containing double bond, ring structure or aromatic structure and connecting linear linker sections. INDEPENDENT CLAIMS are included for the following: (1) formation of electron microscopy grid, which involves providing a perforated substrate, providing a support film on the perforated substrate, attaching linker molecule (I) to the support film via the anchoring group, partially modifying the linker molecules attached to the support film to linker molecules (M2) attached to the support film. The modification of the linker molecules involves chemical or physical, preferably photoisomerization; (2) immobilizing an analyte (16) to an electron microscopy grid, which involves providing an electron microscopy grid, contacting the electron microscopy grid with a solution comprising the analyte for immobilizing the analyte on the electron microscopy grid by the formation of a bond between the binding unit and the analyte, and optionally, partially or fully removing the solution from the electron microscopy grid; (3) determining the structure of the analyte, which involves (a) immobilizing the analyte by providing a primed electron microscopy grid, (b) vitrifying the primed electron microscopy grid, (c) acquiring electron microscopy images from the electron microscopy grid, and (d) reconstructing the structure of the analyte from the electron microscopy images; and (4) linker molecule (I) for attaching the analyte to the support film or the surface. DESCRIPTION OF DRAWING(S) - The drawing shows a schematic view of the electron microscopy grid. Linker molecules (1,2) Support film (4) Perforated substrate (5) Electron microscopy grid (6) Analyte (16)