CRISPR/Cas9 system are widely used in gene editing due to its high efficiency, simple operation and wide species adaptability. The system consists of a guide RNA (sgRNA) that targets the target DNA series and Cas9 with cleavage enzyme activity. In recent years, researchers have developed a range of super-resolution live cell imaging techniques by combining nuclease-inactivated Cas9 mutants dCas9 (dead Cas9) or sgRNA with fluorescent proteins (FPs), organic dyes, and quantum dots (QDs). This technology helps researchers to study different genes, chromosomes and the spatio-temporal relationship between genes and chromosomes at higher resolutions, which is of great significance to promote the rapid development of genetics, cell biology and biomedicine. This paper summarizes the advances in live cell imaging technology based on CRISPR/Cas9 system, which is expected to further expand the wide application in the biomedical field.