Objective In order to make the labeled neurons and glia with 1,1′-dioctadecy-3,3,3′,3,-tetramethylindocarbocyanine perchlorate(DiI) diolistic labeling for long time preservation and ultrastructural study. Methods DiI diotistic assays were done on the sections of visual cortex of 20 C57/B6J mice to label neurons and glia, then the targeted neurons were photoconverted and prepared for ultrathin section. Results The neurons after the photoconversion could still show fine shape with spines on dendrites and decent synapse ultrastructure. Conclusion After DiI diotistic labeling and photoconversation, the labeled neurons can present satisfied morphological details with either light microscopy or electron microscopy. The organelle is well and the synaptic structure is clear either.