The invention belongs to the technical field of biological detection, and particularly discloses a method for immunolocalization of
intracellular protein by a
scanning electron microscope based on nanogold labeling. The method comprises the following steps: collecting cells, cleaning the cells with PBS, and fixing the cells with
paraformaldehyde; heating and melting
agarose, preserving heat, gently resuspending the
cell precipitate, curing and demolding to form a
cell-
agarose complex; embedding the
cell-
agarose complex, quickly freezing,
slicing, attaching to the surface of ITO (
Indium Tin Oxide) conductive glass, and
drying at
room temperature to obtain a slide; cleaning and
drying the slide, and sequentially carrying out primary
antibody incubation and nanogold secondary
antibody incubation; positioning a
soft agar region by adopting an SE mode of a
scanning electron microscope and observing the morphology of the single cell; and detecting the position and the distribution form of the gold nanoparticles by adopting a BSE mode. According to the method, high-resolution, high-fidelity and high-specificity positioning of the
intracellular protein is realized, the efficiency and accuracy of
intracellular protein research are remarkably improved, and the method has important scientific value and application prospect.