Disclosed is an SiO2 nano-particle taking a positive electrical polyelectrolyte as a template and doped with a negative electricity dye, comprising a nuclear shell structure, a kernel comprises a polyelectrolyte cluster with positive electricity, and is doped with the fluorescent dye with negative electricity and SiO2, as well as a selectable additive with negative electricity, and a shell is composed of the SiO2, wherein, the sum of negative charges of the additive and the fluorescent dye is less than the number of positive charges of the polyelectrolyte cluster. A preparation method of the SiO2 nano-particle comprises steps of: composite solution preparation of the polyelectrolyte and the fluorescent dye; formation of the SiO2 nano-particle by hydrolysis and condensation of an organic silicon source; and organic silicon source coating. The invention can be widely applied to the doping of various fluorescent dyes with negative electricity groups in silicon dioxide, and can realize simultaneous doping of a plurality of dyes and control of dye doping amount and particle sizes and appearances of products, the prepared SiO2 nano-particle is spherical, the particle sizes are uniform, the surface functionalization is easy, and the formed colloid has good stability, so as to provide more options for biomedicine, theoretical research and the like.