Phase-selective staining of model and cell membranes, lipid droplets and lipoproteins with fluorescent solvatochromic pyrene probes
| Autor: | Yosuke Niko, Alicia Alonso, Rocío León-Irra, Andrey S. Klymchenko, Félix M. Goñi, Ixone Esnal, Jesús Sot, Bingen G. Monasterio |
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| Přispěvatelé: | European Commission, Ministerio de Ciencia, Innovación y Universidades (España), Agencia Estatal de Investigación (España), Eusko Jaurlaritza, Fundación Biofísica Bizkaia |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Lipoproteins Biophysics CHO Cells 02 engineering and technology Biochemistry Fluorescence spectroscopy Push-pull pyrene probes 03 medical and health sciences chemistry.chemical_compound Cricetulus Lipid droplet Fluorescence microscope Animals Humans Fluorescent Dyes Lipid phases Pyrenes Staining and Labeling Vesicle Erythrocyte Membrane Nile red Lipid Droplets Cell Biology 021001 nanoscience & nanotechnology Fluorescence Confocal fluorescence microscopy 030104 developmental biology Membrane Microscopy Fluorescence chemistry lipids (amino acids peptides and proteins) Giant unilamellar vesicles 0210 nano-technology Laurdan |
| Popis: | 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. This work was supported in part by grants from the Spanish Ministry of Science (FEDER MINECO PGC2018-099857-B-I00) and the Basque Government (grants No. IT1264-19 and IT1270-19), by the Fundación Biofísica Bizkaia and by the Basque Excellence Research Centre (BERC) program of the Basque Government. |
| Databáze: | OpenAIRE |
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