Simultaneous multi-lifetime multi-color STED imaging for colocalization analyses
Authors: Bückers J, Wildanger D, Vicidomini G, Kastrup L, Hell SW
CellNetworks People: Hell Stefan
Journal: Opt Express. 2011 Feb 14;19(4):3130-43. doi: 10.1364/OE.19.003130

We describe a STED microscope optimized for colocalization experiments with up to three colors. Two fluorescence labels are separated by their fluorescence lifetime whereas a third channel is discriminated by the wavelength of fluorescence emission. Since it does not require a second STED beam, separating by lifetime is insensitive to drift and thus optimally suited for colocalization analyses. Furthermore, we propose a setup having a second STED beam for long duration multicolor recording.