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Sphingosine 1-phosphate (S1P) is a central regulatory lipid, produced via the phosphorylation of sphingosine. S1P signals both intra- and extracellularly, binding to the S1P1-5 family of G-protein coupled receptors to control fundamental processes like cell migration, division, and cytoskeletal organization. This highly fluorescent derivative, carrying the tetramethylrhodamine (TMR) label, is perfectly suited for high-resolution live-cell imaging and tracking of S1P internalization and receptor dynamics.