Abstract: Antisense imaging is an important part in the field of nuclear medicine imaging. The method of ⁹⁹Tc^m-labeling oligonucleotides was reviewed in this article based on the preparation of antisense oligonucleotide probes. It mainly involves the chemical modification, the appropriate labeling method of oligonucleotides, the advantages of different kinds of chelators, the optimization of the reaction conditions of labeling and the usage in antisense imaging. A simple programmatic way to synthesize a stable and efficient gene antisense probe will provide new impetus in the development of antisense imaging, and bring the goodness to the diagnosis and treatment of tumors.