The invention discloses a preparation method of mesophase carbon microspheres from coal liquefaction residues serving as raw materials, which comprises the following steps of: firstly carrying out solvent extraction on the coal liquefaction residues, then carrying out a series of processing to obtain refined asphalt with different contents of quinoline insoluble substances, and carrying out thermal polycondensation reaction on the refined asphalt in the presence of a chemical auxiliary agent and a nucleating accelerator at the temperature of 380-450 DEG C, the pressure of 0-5 MPa and the stirring rotation speed of 40-500 r/min for 1-20 hours to obtain the mesophase microspheres. According to the invention, the chemical auxiliary agent and the nucleating accelerator can be added during theheat treatment process to improve the yield and narrow the particle size distribution of the mesophase carbon microbeads, and the mesophase carbon microspheres are prepared from the coal liquefactionresidues serving as the raw materials and have the advantages of low cost, simple preparation process, high yield (up to 35%), narrow particle size distribution, good sphericity, etc.