Journal
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES
Volume 1863, Issue 1, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.bbamem.2020.183470
Keywords
Push-pull pyrene probes; Confocal fluorescence microscopy; Fluorescence spectroscopy; Lipid phases; Giant unilamellar vesicles
Categories
Funding
- Spanish Ministry of Science [FEDER MINECO PGC2018-099857-B-I00]
- Basque Government [IT1264-19, IT1270-19]
- Fundacion Biofisica Bizkaia
- Basque Excellence Research Centre (BERC) program of the Basque Government
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The push-pull solvatochromic pyrene derivatives PA and PK have been successfully applied as polarity-sensitive probes in the study of model membrane vesicles, cells and purified human serum lipoproteins. These probes provide information about lipid order in biomembranes, with the advantage of being excitable by a standard 405 nm laser light. The results from the study showed the capability of PA and PK to report differently on liquid-disordered, liquid-ordered and gel phase bilayers.
The push-pull solvatochromic pyrene derivatives PA and PK have been applied to the study of model membrane vesicles, cells and purified human serum lipoproteins, using both confocal fluorescence microscopy and fluorescence spectroscopy. These polarity-sensitive probes provide information similar to that obtained by Laurdan or Prodan, i.e. mainly lipid order in biomembranes, but they have the essential advantage of being excitable by a standard 405 nm laser light, bypassing the use of multiphoton excitation. In addition, they are brighter and much more photostable than those dimethylamino naphthalene derivatives. Our results with model membrane spectroscopy (multilamellar vesicles) and with microscopy (giant unilamellar vesicles) showed the capacity of PA and PK to report differently on liquid-disordered, liquid-ordered and gel phase bilayers. Moreover, a ratiometric parameter, the Red/Blue Intensity Ratio (RBIR) could be used for inter-domain, inter-vesicle and even inter-technique comparison, and the appropriate microscopy-spectroscopy conversion coefficients could be estimated. In studies at the cellular level, PA probe stained almost exclusively the plasma membrane of red blood cells, revealing its high degree of lipid order. Using Chinese Hamster Ovary cells PA was shown to be an excellent probe for the detection of cytoplasmic lipid droplets, superior to Nile Red in that PA provides simultaneously a detailed information of membrane order in the whole cell, in which the lipid droplets appear with a very good contrast. Moreover, spectrofluorometric data of PA-stained serum lipoproteins indicated an essentially identical value of RBIR for lipid droplets and for high-density lipoproteins.
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