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Proceedings Paper

FRET-sensor for imaging with lifetime resolution
Author(s): Alexander L. Rusanov; Tatiana V. Ivashina; Leonid M. Vinokurov; Alexander S. Goryashenko; Victoria V. Zherdeva; Alexander P. Savitsky
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Paper 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.

Paper Details

Date Published: 25 November 2010
PDF: 6 pages
Proc. SPIE 7376, Laser Applications in Life Sciences, 737611 (25 November 2010); doi: 10.1117/12.871367
Show Author Affiliations
Alexander L. Rusanov, M.V. Lomonosov Moscow State Univ. (Russian Federation)
A.N. Bach Institute of Biochemistry (Russian Federation)
Tatiana V. Ivashina, Skryabin Institute of Biochemistry and Physiology (Russian Federation)
Leonid M. Vinokurov, Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry (Russian Federation)
Alexander S. Goryashenko, M.V. Lomonosov Moscow State Univ. (Russian Federation)
A.N. Bach Institute of Biochemistry (Russian Federation)
Victoria V. Zherdeva, A.N. Bach Institute of Biochemistry (Russian Federation)
Alexander P. Savitsky, M.V. Lomonosov Moscow State Univ. (Russian Federation)
A.N. Bach Institute of Biochemistry (Russian Federation)


Published in SPIE Proceedings Vol. 7376:
Laser Applications in Life Sciences
Matti Kinnunen; Risto Myllylä, Editor(s)

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