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24 November 2010 FRET-sensor for imaging with lifetime resolution
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Proceedings Volume 7376, Laser Applications in Life Sciences; 737611 (2010)
Event: Laser Applications in Life Sciences 2010, 2010, Oulu, Finland
FRET-sensor with nonfluorescent protein as an acceptor was synthesized to observe caspase-3 activity in lifetime mode. We inserted caspase-3 cleavable linker between red highly fluorescent protein TagRFP and chromoprotein KFP. Dynamic light scattering was used to determine size of the fusion protein. Incubation with caspase-3 lead to increase both fluorescence intensity and lifetime of the construction. Cleavage of the linker between proteins was confirmed by electrophoresis and immunoblotting. FLIM-microscopy showed the differences between fluorescence decays of A549 cell line expressed TagRFP and TagRFP-23-KFP.
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Alexander L. Rusanov, Tatiana V. Ivashina, Leonid M. Vinokurov, Alexander S. Goryashenko, Victoria V. Zherdeva, and Alexander P. Savitsky "FRET-sensor for imaging with lifetime resolution", Proc. SPIE 7376, Laser Applications in Life Sciences, 737611 (24 November 2010);

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