Imaging is a valuable visual tool for scientific research, providing a qualitative measure of analysis. There are various methods for mitochondrial fluorescence imaging, such as small molecule dyes and fluorescent proteins. Small molecule dyes are frequently used in fluorescence microscopy; however, they have low specificity, high toxicity, and photo-bleach easily. By designing a bicistronic plasmid containing two fluorescent protein tagging sites, an efficient method for mitochondrial fluorescence imaging can be utilized.