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Imaging intracellular viscosity by a new molecular rotor suitable for phasor analysis of fluorescence lifetime

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Abstract

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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Notes

  1. Note that HP mixtures contained always at least 80 % glycerol.

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Acknowledgments

We thank Dr. Marco Cecchini and Prof. Enrico Gratton for useful discussions. This work was partially supported by the Italian Ministry for University and Research (MiUR) under the framework of the FIRB project RBPR05JH2P and PRIN project 2010BJ23MN_004 and by the European Union Seventh Framework Programme (FP7/2007–2013) under grant agreement no. NMP4-LA-2009-229289 NanoII and grant agreement no. NMP3-SL-2009-229294 NanoCARD.

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Authors and Affiliations

  1. NEST, Scuola Normale Superiore and Istituto Nanoscienze, CNR, Piazza San Silvestro 12, 56127, Pisa, Italy

    Antonella Battisti, Silvio Panettieri, Gerardo Abbandonato, Emanuela Jacchetti, Fabio Beltram & Ranieri Bizzarri

  2. Center for Nanotechnology Innovation at NEST, Istituto Italiano di Tecnologia, Piazza San Silvestro 12, 56127, Pisa, Italy

    Antonella Battisti, Gerardo Abbandonato, Francesco Cardarelli & Giovanni Signore

  3. Istituto di Biofisica, CNR, via Moruzzi 1, 56124, Pisa, Italy

    Ranieri Bizzarri

Authors
  1. Antonella Battisti
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  2. Silvio Panettieri
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  3. Gerardo Abbandonato
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  4. Emanuela Jacchetti
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  5. Francesco Cardarelli
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  6. Giovanni Signore
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  7. Fabio Beltram
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  8. Ranieri Bizzarri
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Corresponding authors

Correspondence to Giovanni Signore or Ranieri Bizzarri.

Additional information

Published in the topical collection Optical Nanosensing in Cells with guest editor Francesco Baldini.

Antonella Battisti and Silvio Panettieri contributed equally to this work.

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Battisti, A., Panettieri, S., Abbandonato, G. et al. Imaging intracellular viscosity by a new molecular rotor suitable for phasor analysis of fluorescence lifetime. Anal Bioanal Chem 405, 6223–6233 (2013). https://doi.org/10.1007/s00216-013-7084-x

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  • DOI: https://doi.org/10.1007/s00216-013-7084-x

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