Bioorthogonal Reaction Pairs Enable Simultaneous, Selective, Multi-Target Imaging


  • The authors would like to thank Dr. Jack Szostak at MGH for the use of the stopped-flow spectrometer. The authors would also like to thank Alex Chudnovskiy, Elizabeth Tiglao, and Yoshi Iwamoto of CSB for technical assistance with cell culture and flow cytometry as well as Neal K. Devaraj and Jason R. McCarthy for helpful discussions. This research is supported in part by NIH grants RO1EB010011 and P50A86355.


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Mutually orthogonal tetrazine–trans-cyclooctene and azide–cyclooctyne cycloaddition reactions were used simultaneously for the bioorthogonal labeling of two different live cell populations in the same culture (see scheme). These small-molecule probes show good chemical reactivity and can be readily incorporated into biological systems.