Fluorescence lifetime imaging for viscosity and diffusion measurements

Klaus Suhling; Yurema Teijeiro-Gonzalez; I. Emilie Steinmark; Arjuna L. James; Augoustina M. Economou; Gokhan Yahioglu; Alix Le Marois; Liisa M. Hirvonen; Jakub Nedbal; James A. Levitt; Pei-Hua Chung; Cécile A. Dreiss; Andrew J. Beavil; Rebecca L. Beavil; Elena Ortiz-Zapater; Christian D. Lorenz; Maddy Parsons; Alessandro Crnjar; Bethan Cornell; Carla Molteni

doi:10.1117/12.2508744

22 February 2019 Fluorescence lifetime imaging for viscosity and diffusion measurements

Klaus Suhling, Yurema Teijeiro-Gonzalez, I. Emilie Steinmark, Arjuna L. James, Augoustina M. Economou, Gokhan Yahioglu, Alix Le Marois, Liisa M. Hirvonen, Jakub Nedbal, James A. Levitt, Pei-Hua Chung, Cécile A. Dreiss, Andrew J. Beavil, Rebecca L. Beavil, Elena Ortiz-Zapater, Christian D. Lorenz, Maddy Parsons, Alessandro Crnjar, Bethan Cornell, Carla Molteni

Author Affiliations +

Proceedings Volume 10882, Multiphoton Microscopy in the Biomedical Sciences XIX; 1088210 (2019) https://doi.org/10.1117/12.2508744
Event: SPIE BiOS, 2019, San Francisco, California, United States

Abstract

Imaging viscosity and its spatiotemporal patterns can provide valuable insight into the underlying physical conditions of biochemical reactions and biological processes in cells and tissues. One way to measure viscosity and diffusion is the use of fluorescence recovery after photobleaching (FRAP). We combine FRAP with FLIM and time-resolved fluorescence anisotropy imaging (tr-FAIM), by acquiring time- and polarization-resolved fluorescence images in every frame of a FRAP series. This allows us to simultaneously monitor translational and rotational diffusion. This approach can be applied to measuring diffusion in homogeneous and heterogeneous environments, and in principle also allows the study of homo-FRET. Another way to measure viscosity and diffusion is through specific flexible dyes, e.g. fluorescent molecular rotors, whose fluorescence quantum yield and fluorescence lifetime depend on the viscosity of the environment, in combination with fluorescence lifetime imaging (FLIM). We show that a bodipybased fluorescent molecular rotor targeting mitochondria reports on their viscosity, which changes under physiological stimuli. Both methods can optically measure viscosity and diffusion on the micrometer scale.

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Klaus Suhling, Yurema Teijeiro-Gonzalez, I. Emilie Steinmark, Arjuna L. James, Augoustina M. Economou, Gokhan Yahioglu, Alix Le Marois, Liisa M. Hirvonen, Jakub Nedbal, James A. Levitt, Pei-Hua Chung, Cécile A. Dreiss, Andrew J. Beavil, Rebecca L. Beavil, Elena Ortiz-Zapater, Christian D. Lorenz, Maddy Parsons, Alessandro Crnjar, Bethan Cornell, and Carla Molteni "Fluorescence lifetime imaging for viscosity and diffusion measurements", Proc. SPIE 10882, Multiphoton Microscopy in the Biomedical Sciences XIX, 1088210 (22 February 2019); https://doi.org/10.1117/12.2508744

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