Characterization of a new series of fluorescent probes for imaging membrane order

PLoS One. 2013;8(2):e52960. doi: 10.1371/journal.pone.0052960. Epub 2013 Feb 4.


Visualization and quantification of lipid order is an important tool in membrane biophysics and cell biology, but the availability of environmentally sensitive fluorescent membrane probes is limited. Here, we present the characterization of the novel fluorescent dyes PY3304, PY3174 and PY3184, whose fluorescence properties are sensitive to membrane lipid order. In artificial bilayers, the fluorescence emission spectra are red-shifted between the liquid-ordered and liquid-disordered phases. Using ratiometric imaging we demonstrate that the degree of membrane order can be quantitatively determined in artificial liposomes as well as live cells and intact, live zebrafish embryos. Finally, we show that the fluorescence lifetime of the dyes is also dependent on bilayer order. These probes expand the current palate of lipid order-sensing fluorophores affording greater flexibility in the excitation/emission wavelengths and possibly new opportunities in membrane biology.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 2-Naphthylamine / analogs & derivatives*
  • 2-Naphthylamine / analysis
  • 2-Naphthylamine / chemical synthesis
  • Animals
  • Cholesterol / chemistry*
  • Embryo, Nonmammalian
  • Fluorescent Dyes / analysis*
  • Fluorescent Dyes / chemical synthesis
  • HeLa Cells
  • Humans
  • Lipid Bilayers / chemistry*
  • Liposomes / chemistry*
  • Membrane Microdomains / chemistry*
  • Phosphatidylcholines / chemistry*
  • Pyridines / analysis*
  • Pyridines / chemical synthesis
  • Quaternary Ammonium Compounds / analysis*
  • Quaternary Ammonium Compounds / chemical synthesis
  • Quinolines / analysis*
  • Quinolines / chemical synthesis
  • Spectrometry, Fluorescence
  • Sphingomyelins / chemistry*
  • Zebrafish


  • Fluorescent Dyes
  • Lipid Bilayers
  • Liposomes
  • Phosphatidylcholines
  • Pyridines
  • Quaternary Ammonium Compounds
  • Quinolines
  • Sphingomyelins
  • palmitoylsphingomyelin
  • Cholesterol
  • 2-Naphthylamine
  • 1,2-oleoylphosphatidylcholine