title: Characterization of a new series of fluorescent probes for imaging membrane order.
abstract: 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.
authors: Centre for Vascular Research and Australian Centre for Nanomedicine, University of New South Wales, Sydney, Australia.
authors: Joanna M
authors: JM
authors: Kwiatek
authors: Dylan M
authors: DM
authors: Owen
authors: Ahmed
authors: A
authors: Abu-Siniyeh
authors: Ping
authors: P
authors: Yan
authors: Leslie M
authors: LM
authors: Loew
authors: Katharina
authors: K
authors: Gaus
mesh_terms: D015081:2-Naphthylamine / Q000031:analogs & derivatives* / Q000032:analysis / Q000138:chemical synthesis; D000818:Animals; D002784:Cholesterol / Q000737:chemistry*; D004625:Embryo, Nonmammalian; D005456:Fluorescent Dyes / Q000032:analysis* / Q000138:chemical synthesis; D006367:HeLa Cells; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry*; D021962:Membrane Microdomains / Q000737:chemistry*; D010713:Phosphatidylcholines / Q000737:chemistry*; D011725:Pyridines / Q000032:analysis* / Q000138:chemical synthesis; D000644:Quaternary Ammonium Compounds / Q000032:analysis* / Q000138:chemical synthesis; D011804:Quinolines / Q000032:analysis* / Q000138:chemical synthesis; D013050:Spectrometry, Fluorescence; D013109:Sphingomyelins / Q000737:chemistry*; D015027:Zebrafish
chemical_list: D005456:Fluorescent Dyes; D008051:Lipid Bilayers; D008081:Liposomes; D010713:Phosphatidylcholines; D011725:Pyridines; D000644:Quaternary Ammonium Compounds; D011804:Quinolines; D013109:Sphingomyelins; C033171:palmitoylsphingomyelin; D002784:Cholesterol; D015081:2-Naphthylamine; C017251:1,2-oleoylphosphatidylcholine
references: Virology. 2009 Mar 30;386(1):192-202
references: 19185899
references: J Lipid Res. 2006 Jul;47(7):1597-8
references: 16645198
references: Biophys J. 1990 Jun;57(6):1179-86
references: 2393703
references: Biophys J. 2009 May 6;96(9):3461-70
references: 19413953
references: Proc Natl Acad Sci U S A. 2009 Sep 29;106(39):16645-50
references: 19805351
references: Nat Protoc. 2011 Dec 08;7(1):24-35
references: 22157973
references: Biophys J. 1997 Jun;72(6):2413-29
references: 9168019
references: Biophys J. 2003 Oct;85(4):2406-16
references: 14507704
references: FEBS Lett. 2007 May 22;581(11):2098-104
references: 17382322
references: Biophys J. 1991 Jul;60(1):179-89
references: 1883937
references: J Cell Biol. 2005 Oct 10;171(1):121-31
references: 16203859
references: Biophys J. 2006 Jun 1;90(11):L80-2
references: 16617080
references: Chembiochem. 2007 Mar 26;8(5):553-9
references: 17300111
references: Proc Natl Acad Sci U S A. 2003 Dec 23;100(26):15554-9
references: 14673117
references: Nat Rev Mol Cell Biol. 2010 Oct;11(10):688-99
references: 20861879
references: Semin Cell Dev Biol. 2007 Oct;18(5):591-8
references: 17728161
references: Biophys J. 2005 Jul;89(1):L04-6
references: 15879475
references: Nat Methods. 2010 Nov;7(11):865-6
references: 20881966
references: Mol Membr Biol. 2010 Aug;27(4-6):178-89
references: 20540668
references: Biochemistry. 1979 Jul 10;18(14):3075-8
references: 465454
references: J Cell Biol. 1987 Oct;105(4):1623-35
references: 3667693
references: Biochim Biophys Acta. 2006 Oct;1758(10):1541-56
references: 16854370
references: J Biol Chem. 1999 Jan 22;274(4):2038-44
references: 9890962
references: Biophys J. 2006 Apr 1;90(7):2563-75
references: 16415047
references: J Immunol. 2011 Mar 15;186(6):3505-16
references: 21307290
references: Biophys J. 2010 Jul 7;99(1):L7-9
references: 20655825
references: Microsc Res Tech. 2010 Jun;73(6):618-22
references: 19937746
references: Annu Rev Phys Chem. 2005;56:309-36
references: 15796703
references: Traffic. 2009 Aug;10(8):962-71
references: 19416480
references: Science. 2010 Jan 1;327(5961):46-50
references: 20044567
references: J Neurosci Methods. 2004 Apr 30;134(2):179-90
references: 15003384
references: J Biomed Opt. 2010 Jul-Aug;15(4):046017
references: 20799819
references: Nature. 1997 Jun 5;387(6633):569-72
references: 9177342