Phase-selective staining of model and cell membranes, lipid droplets and lipoproteins with fluorescent solvatochromic pyrene probes

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...

Full description

Saved in:
Bibliographic Details
Published in:Biochimica et biophysica acta. Biomembranes 2021-01, Vol.1863 (1), p.183470-183470, Article 183470
Main Authors: Sot, Jesús, Esnal, Ixone, Monasterio, Bingen G., León-Irra, Rocío, Niko, Yosuke, Goñi, Félix M., Klymchenko, Andrey, Alonso, Alicia
Format: Article
Language:English
Subjects:
Animals
Biochemistry, Molecular Biology
CHO Cells
Confocal fluorescence microscopy
Cricetulus
Erythrocyte Membrane - chemistry
Erythrocyte Membrane - metabolism
Fluorescence spectroscopy
Fluorescent Dyes - chemistry
Fluorescent Dyes - pharmacology
Giant unilamellar vesicles
Humans
Life Sciences
Lipid Droplets - chemistry
Lipid Droplets - metabolism
Lipid phases
Lipoproteins - chemistry
Lipoproteins - pharmacology
Microscopy, Fluorescence
Push-pull pyrene probes
Pyrenes - chemistry
Pyrenes - pharmacology
Staining and Labeling
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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. [Display omitted] •The push-pull solvatochromic pyrene derivatives PA and PK have been applied to the study of model and cell membranes.•These polarity-sensitive probes can be excited by a 405 nm laser light, bypassing the use of multiphoton excitation.•PA and PK are brighter and more photostable than the dimethylamino naphthalene probes currently in use.•PA and PK report differently on liquid-disordered, liquid-ordered and gel phase bilayers.•These probes have been used to retrieve information on cell lipid droplets and serum lipoproteins.
ISSN:0005-2736
1879-2642
DOI:10.1016/j.bbamem.2020.183470