Fluorescence liftime imaging (FLIM) of modified hydrophobic bodipy dyes that act as fluorescent molecular rotors shows that the fluorescence lifetime of these probes is a function of the microviscosity of their environment. Incubating cells with these dyes, we find a punctate and continuous distribution of the dye in cells. The viscosity value obtained in what appears to be endocytotic vesicles in living cells is around 100 times higher than that of water and of cellular cytoplasm.Time-resolved fluorescence anisotropy measurements also yield rotational correlation times consistent with large microviscosity values. In this way, we successfully develop a practical and versatile approach to map the microviscosity in cells based on imaging fluorescent molecular rotors.
© 2010 Chinese Optics Letters
(110.0110) Imaging systems : Imaging systems
(170.0170) Medical optics and biotechnology : Medical optics and biotechnology
(180.0180) Microscopy : Microscopy
(300.0300) Spectroscopy : Spectroscopy
James A. Levitt, Marina K. Kuimova, Gokhan Yahioglu, Pei-Hua Chung, Klaus Suhling, and David Phillips, "Fluorescence lifetime imaging of molecular rotors to map microviscosity in cells (Invited Paper)," Chin. Opt. Lett. 8, 926-930 (2010)