Optics Express 2011-02-14

Simultaneous multi-lifetime multi-color STED imaging for colocalization analyses.

Johanna Bückers, Dominik Wildanger, Giuseppe Vicidomini, Lars Kastrup, Stefan W Hell

Index: Opt. Express 19(4) , 3130-43, (2011)

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Abstract

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.