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.
Department of Radiology, Washington University in St. Louis School of Medicine, Saint Louis, MO 63110, USA.
Tiffany P
TP
Gustafson
Qian
Q
Cao
Samuel
S
Achilefu
Mikhail Y
MY
Berezin
D000818:Animals; D002645:Chickens; D005454:Fluorescence Polarization / Q000379:methods; D005456:Fluorescent Dyes / Q000737:chemistry*; D007074:Immunoglobulin G / Q000737:chemistry; D007211:Indoles / Q000737:chemistry*; D009113:Muramidase / Q000737:chemistry; D051381:Rats; D012710:Serum Albumin, Bovine / Q000737:chemistry; D019265:Spectroscopy, Near-Infrared / Q000379:methods
D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D007211:Indoles; D012710:Serum Albumin, Bovine; C098209:VPM chloride; D009113:Muramidase