Fluorescent Lipids

Avanti offers a wide variety of fluorescent lipids to fulfill your research needs. There are several moieties that we use to install fluorescent properties in our lipids. As always, these lipids are made with the same quality standards as their non-fluorescent counterparts. The following moieties have been used to install fluorescent properties in phospholipids, sphingolipids, sterols, glycerolipids, and others: nitrobenzoxadiazole (NBD), 1,6-diphenyl-1,3,5-hexatriene (DPH), pyrenedecanoyl, cyanine 5, and TopFluor. To find out more about each type of fluorescent moiety see the information below!

Nitrobenzoxadiazole (NBD) is a common fluorescent label for lipids. NBD was discovered several decades ago but continues to be a popular choice for fluorescent labeling of lipids due to its desirable fluorescence characteristics. NBD-labeled lipids offer a wide array of applications in membrane and cell biology. NBD-labeled lipids, specifically NBD-PE, have been shown to have maximum excitation and emission wavelengths of 465nm and 535nm, respectively.1,2

1,6-diphenyl-1,3,5-hexatriene (DPH) has become a popular moiety for investigating the dynamic and structural properties of lipid bilayers and cellular membranes. The technique used for these studies is steady-state or time-resolved fluorescence anisotropy. DPH-labels help explore properties such as lipid tail order, hydration at the membrane-water interface, membrane electrostatic properties, and dynamics of membrane components.3

Pyrenedecanoyl is of the most common fluorescent labels and is commonly incorporated as a head group in phospho- and glycerophospholipids. Pyrene labeled lipids are particularly useful for monitoring membrane fusion and phospholipid transfer processes due to their excimer-forming properties. The formation of the excited state pyrene-dimers, also called excimers, is concentration-dependent, and exhibit a red-shifted emission peak at ~470nm.4

Cyanine 5 (Cy5) are among the most used oligonucleotide labels for microarray experiments, fluorescent in situ hybridization (FISH), real-time PCR (RT-PCR), and FRET studies. This label is considered to be environmentally insensitive. This moiety is made of two indole rings connected by five carbon polymethine bridge. This bridge can undergo cis-trans isomerization from the first excited single state which competes with fluorescence. If the rate of cis-trans isomerization is reduced or eliminated they produce greater fluorescence.5

TopFluor-labeled lipids are great aids in studying the distribution of lipids, lipid diffusion rates, and lipid dynamics. TopFluor has been used to study accessibility in the lipid bilayer, cellular localization, and membrane dynamics via fluorescence recovery. Studies have indicated that this fluorescent label alters the biochemical and biophysical properties of lipids less than some other fluorescent labels such as NBD.6

  1. Haldar S., Chattopadhyay A. (2012) Application of NBD-Labeled Lipids in Membrane and Cell Biology. In: Mély Y., Duportail G. (eds) Fluorescent Methods to Study Biological Membranes. Springer Series on Fluorescence (Methods and Applications), vol 13. Springer, Berlin, Heidelberg. https://doi.org/10.1007/4243_2012_43
  2. Monti J.A., Christian, S.T., Shaw, W.A. (1978) Synthesis and properties of a highly fluorescent derivative of phosphatidylethanolamine. J. Lip. Res. 19:222-228.
  3. Chetan Poojari, Natalia Wilkosz, Rafael B. Lira, Rumiana Dimova, Piotr Jurkiewicz, Rafał Petka, Mariusz Kepczynski, Tomasz Róg, Behavior of the DPH fluorescence probe in membranes perturbed by drugs, Chemistry and Physics of Lipids, Volume 223, 2019, 04784, ISSN 0009-3084, https://doi.org/10.1016/j.chemphyslip.2019.104784.
  4. Molecular Probes Handbook: A guide to fluorescent probes and labeling technologies (11th ed.) Chapter 13: Probes for Lipids and Membranes.
  5. Kretschy N., Sack M., Somoza, M. (2016) Sequence-Dependent Fluorescence of Cy3- and Cy5-Labeled Double-Stranded DNA. Bioconjug. Chem. 27:840-848
  6. Shirey, C. M., Ward, K. E., & Stahelin, R. V. (2016). Investigation of the biophysical properties of a fluorescently modified ceramide-1-phosphate. Chemistry and physics of lipids, 200, 32–41. https://doi.org/10.1016/j.chem...