Probing Polarity and Heterogeneity of Lipid Droplets in Live Cells Using a Push-Pull Fluorophore

被引:120
作者
Collot, Mayeul [1 ]
Bou, Sophie [1 ]
Fam, Tkhe Kyong [1 ]
Richert, Ludovic [1 ]
Mely, Yves [1 ]
Danglot, Lydia [2 ,3 ]
Klymchenko, Andrey S. [1 ]
机构
[1] Univ Strasbourg, CNRS, Lab Biophoton & Pathol, Fac Pharm,UMR 7021, 74 Route Rhin, F-67401 Illkirch Graffenstaden, France
[2] Univ Paris Diderot, Inst Jacques Monod, CNRS, Sorbonne Paris Cite,UMR 7592, F-75013 Paris, France
[3] INSERM, Inst Psychiat & Neurosci Paris, Membrane Traff Hlth & Diseased Brain, U894, 102 Rue Sante, F-75014 Paris, France
关键词
NILE RED; FLUORESCENCE; PROBES; DYES; BRIGHTNESS; BIOPROBE; DESIGN; ORDER; MCF7;
D O I
10.1021/acs.analchem.8b04218
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Lipid droplets (LDs) are organelles composed of a lipid core surrounded by a phospholipid monolayer. Lately, LDs have attracted considerable attention due to recent studies demonstrating their role in a variety of physiological processes as well as diseases. Herein we synthesized a push-pull molecule named DAF (Dimethyl Aniline Furaldehyde) that possesses a strong positive solvatochromism in emission of 119 nm from toluene to methanol. Its impressive fluorogenic properties from water to oil (2000-fold) as well as its high quantum yields (up to 0.97) led us to investigate its ability to sense the distribution of polarity in live cells by fluorescence ratiometric imaging. When added to live cells and excited at 405 nm, DAF immediately and brightly stain lipid droplets using a blue channel (410-500 nm) and cytoplasm in a red channel (500-600 nm). DAF also proved to be compatible with fixation thus allowing 3D imaging of LDs in their cytoplasm environment. Taking advantage of DAF emission in two distinct channels, ratiometric imaging was successfully performed and led to the polarity mapping of the cell unraveling some heterogeneity in polarity within LDs of the same cell.
引用
收藏
页码:1928 / 1935
页数:8
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