▎ 摘　　要

NOVELTY - A single molecule fluorescence observation method involves utilizing thin layer of graphene nano metal bonded fluorescence quenching array, and processing single molecule fluorescence observation area surface. Preparation of a graphene substrate on ordered array of nano metals is performed by spin coating para-methoxymethamphetamine, drawing thin layer on graphene layer, and forming nanostructure array. The array is soaked in chemical solution, precooled and deposited by metallic layer followed by subjecting to electron microscope photograph. USE - Method for observation of single molecule fluorescence. ADVANTAGE - The method enables observing single molecule fluorescence by stimulating light with excellent permeability and reduced loss of signal. DETAILED DESCRIPTION - A single molecule fluorescence observation method involves utilizing thin layer of graphene nano metal bonded fluorescence quenching array, where the array comprises a substrate with thin layer graphene combined with metal nano arrays and quenching fluorescent molecules non-specifically bonded without affecting another modification of nano metals, and processing single molecule fluorescence observation area surface by utilizing glass sheet, cleaning surface with a mixture of concentrated sulfuric acid and hydrogen peroxide, blowing high purity nitrogen gas on the sheet, and utilizing transparent adhesive tape for transferring graphene to a glass surface, where the graphene is a thin layer graphene with thickness of 3 atomic layers or less. Preparation of graphene substrate on ordered array of nano metals by spin coating para-methoxymethamphetamine in slide surface, drawing a thin layer on graphene layer with an aluminized electron-beam by evaporation method under an optical microscope, and forming nanostructure array after exposing electron beam. The array is soaked in chemical solution to remove aluminum, where the chemical solution comprises positive resistance developing agent and deionized water mixture in a weight ratio of 1:3-1:5. The array is precooled to 0-4 degrees C for 30-40 minutes, where the developing agent comprises methyl isobutyl ketone (MIBK) and isopropyl alcohol (IPA) in a weight ratio of 1:2-1:4. The array is deposited by a metallic layer by electron beam method, where the metal layer has thickness of 15-20 nm, soaked in acetone, excess metal film is removed by ultrasonic process, followed by subjecting to electron microscope photograph to determine the array integrity.