Defining a polymethine dye for fluorescence anisotropy applications in the near-infrared spectral range

Fluorescence anisotropy in the near-infrared (NIR) spectral range is challenging because of the lack of appropriate NIR fluorescent labels. We have evaluated polymethine fluorescent dyes to identify a leading candidate for NIR anisotropy applications. The NIR dye LS601 demonstrated low fluorescence anisotropy values (r) as a result of its relatively long fluorescent lifetime 1.3 ns. The r value of LS601 unbound and coupled to biological macromolecules was found to have a sufficient dynamic range from 0.24 to 0.37, demonstrating the feasibility of fluorescence anisotropy in the NIR. The viability of fluorescence anisotropy using a NIR label was demonstrated by characterization of dye-protein conjugates. These results open the door to a number of applications in drug discovery, fluorescence anisotropy imaging and contrast agent development. This dye does it! Fluorescence anisotropy in the near-infrared (NIR) spectral range is challenging because appropriate NIR fluorescent labels still need to be identified. Polymethine fluorescent dyes are evaluated for this purpose. The most promising label (LS601) is then utilized in conjugation reactions to demonstrate the feasibility of fluorescence anisotropy in the NIR range (see picture).