Some mRNAs localize to specific regions within eukaryotic cells to express their functions. The movement and localization of mRNA molecules provides valuable information about how they concentrate to particular regions. Recent technical advances in optical microscopy and image analysis algorithms enable real-time tracking of single mRNA molecules in living cells. This chapter presents the methods to visualize and track single β-actin mRNA molecules that localize at the leading edge of chicken embryo fibroblasts. Furthermore, this chapter presents an analysis approach for single-molecule tracking data to extract quantitative information about the microenvironments of the mRNA molecules.