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Synchrotron UV fluorescence microscopy for determining membrane fluidity modification of single bacteria with temperatures

Abstract : Cryopreservation of bacteria requires specific cells pre-adaptation as well as controlled freezing protocol to minimize membrane damage resulting from osmotic cell dehydration. Membrane fluidity has an important role in biophysical events taking place at subzero temperatures, by facilitating or not the exchanges between intracellular and extracellular media. An original approach using Synchrotron fluorescence polarization microscopy was developed to measure membrane fluidity of single bacteria. Cells were labelled with the cytoplasmic membrane probe trimethylammoniumdiphenylhexatriene (TMA-DPH). The degrees of liberty of TMA-DPH inside the bilayer provide a direct marker of membrane fluidity under deep UV excitation. Couples of fluorescence polarization images from zero to forty degrees were recorded by inserting polarizers into the excitation and emission paths. The fluorescence anisotropy of the images was calculated thus allowing the assessment of the cell membrane fluidity. Results indicate intercellular and intracellular heterogeneities with subdomains appearing at low temperature for freeze-sensitive cells.
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https://hal.inrae.fr/hal-02629450
Déposant : Migration Prodinra <>
Soumis le : mercredi 27 mai 2020 - 03:51:00
Dernière modification le : samedi 27 juin 2020 - 19:44:07

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Stéphanie Passot, Frederic Jamme, M. Refregiers, Julie Gautier, Stéphanie Cenard, et al.. Synchrotron UV fluorescence microscopy for determining membrane fluidity modification of single bacteria with temperatures. Biomedical Spectroscopy and Imaging, 2014, 3 (3), pp.203-210. ⟨10.3233/BSI-140062⟩. ⟨hal-02629450⟩

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