Journal
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 405, Issue 19, Pages 6223-6233Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-013-7084-x
Keywords
Intracellular viscosity; Molecular rotor; Phasor approach; FLIM; Mitochondria
Funding
- Italian Ministry for University and Research (MiUR) [RBPR05JH2P, 2010BJ23MN_004]
- European Union [NMP4-LA-2009-229289 NanoII, NMP3-SL-2009-229294 NanoCARD]
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The arsenal of fluorescent probes tailored to functional imaging of cells is rapidly growing and benefits from recent developments in imaging strategies. Here, we present a new molecular rotor, which displays strong absorption in the green region of the spectrum, very little solvatochromism, and strong emission sensitivity to local viscosity. The emission increase is paralleled by an increase in emission lifetime. Owing to its concentration-independent nature, fluorescence lifetime is particularly suitable to image environmental properties, such as viscosity, at the intracellular level. Accordingly, we demonstrate that intracellular viscosity measurements can be efficiently carried out by lifetime imaging with our probe and phasor analysis, an efficient method for measuring lifetime-related properties (e.g., bionalyte concentration or local physicochemical features) in living cells. Notably, we show that it is possible to monitor the partition of our probe into different intracellular regions/organelles and to follow mitochondrial de-energization upon oxidative stress.
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