FRET-sensor for imaging with lifetime resolution

Alexander L. Rusanov; Tatiana V. Ivashina; Leonid M. Vinokurov; Alexander S. Goryashenko; Victoria V. Zherdeva; Alexander P. Savitsky

doi:10.1117/12.871367

24 November 2010 FRET-sensor for imaging with lifetime resolution

Alexander L. Rusanov, Tatiana V. Ivashina, Leonid M. Vinokurov, Alexander S. Goryashenko, Victoria V. Zherdeva, Alexander P. Savitsky

Author Affiliations +

Proceedings Volume 7376, Laser Applications in Life Sciences; 737611 (2010) https://doi.org/10.1117/12.871367
Event: Laser Applications in Life Sciences 2010, 2010, Oulu, Finland

Abstract

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.

Citation Download Citation

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); https://doi.org/10.1117/12.871367

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