The invention discloses a multi-scale uniform and single-dispersion magnetic microsphere and a preparation method of the multi-scale uniform and single-dispersion magnetic microsphere. The multi-scale uniform and single-dispersion magnetic microsphere comprises spinel structure ferrite doped with transition metal elements, the microsphere is a polycrystal micro-nano sphere which is formed by nanocrystalline in an aggregation mode, the diameter of the microsphere ranges from 20nm to 1.23 micrometers, the specific surface area of the microsphere ranges from 0.94 m<2>*g<-1> to 17.31 m<2>*g<-1>, and the size of the nanocrystalline ranges from 9.2nm to 25.8nm. A mixed solvothermal method is adopted, and the size and the composition of particles can be adjusted through adjustment of the volume fraction of water or the ratio of iron to other transition metal salt. According to the preparation method, raw materials are low in price, and can be easily obtained, the cost is low, the technology is simple, the requirement for equipment is low, efficiency is high, and popularization is easy; the multi-scale uniform and single-dispersion magnetic microsphere has the advantages that single dispersion and uniformity are good, and the size and the composition can be adjusted; the multi-scale uniform and single-dispersion magnetic microsphere can have the broad application prospect in the fields such as the magneto-rheological field, the magnetic separation field, the catalysis field, the electrode material field, the pigment field, the high-density magnetic recording material field and the medical diagnosis field.