Spectral phasor analysis of LAURDAN fluorescence in live A549 lung cells to study the hydration and time evolution of intracellular lamellar body-like structures.

Leonel Malacrida 1, 2, 3 Soledad Astrada 1 Arturo Briva 2 Mariela Bollati-Fogolín 1 Enrico Gratton 3 Luis A Bagatolli 4
1 Institut Pasteur de Montevideo
2 UCUR - Universidad de la República [Montevideo]
3 laboratory for fluorescence dynamics
4 MEMPHYS
Abstract : Using LAURDAN spectral imaging and spectral phasor analysis we concurrently studied the growth and hydration state of subcellular organelles (lamellar body-like, LB-like) from live A549 lung cancer cells at different post-confluence days. Our results reveal a time dependent two-step process governing the size and hydration of these intracellular LB-like structures. Specifically, a first step (days 1 to 7) is characterized by an increase in their size, followed by a second one (days 7 to 14) where the organelles display a decrease in their global hydration properties. Interestingly, our results also show that their hydration properties significantly differ from those observed in well-characterized artificial lamellar model membranes, challenging the notion that a pure lamellar membrane organization is present in these organelles at intracellular conditions. Finally, these LB-like structures show a significant increase in their hydration state upon secretion, suggesting a relevant role of entropy during this process.
Keywords : Spectral phasor Membrane biophysics Microscopic imaging LAURDAN Lamellar bodies Macromolecular crowding
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Journal articles
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https://hal-riip.archives-ouvertes.fr/pasteur-01498359
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Submitted on : Monday, October 9, 2017 - 4:45:36 PM
Last modification on : Friday, August 23, 2019 - 3:20:03 PM

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Leonel Malacrida, Soledad Astrada, Arturo Briva, Mariela Bollati-Fogolín, Enrico Gratton, et al.. Spectral phasor analysis of LAURDAN fluorescence in live A549 lung cells to study the hydration and time evolution of intracellular lamellar body-like structures.. BBA - Biochimica et Biophysica Acta, Elsevier, 2016, 1858 (11), pp.2625-2635. ⟨10.1016/j.bbamem.2016.07.017⟩. ⟨pasteur-01498359⟩

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