Diffusion of lipid-like single-molecule fluorophores in the cell membrane.
The dicyanomethylenedihydrofuran (DCDHF) class of single-molecule fluorophores contains an amine donor and a dicyanomethylenedihydrofuran acceptor linked by a conjugated unit (benzene, naphthalene, or styrene). Molecules in this class have a number of useful properties in addition to those usually required for single-molecule studies (such as high fluorescence quantum yield and photostability), including second-order optical nonlinearity, large ground-state dipole moment, and sensitivity to local environment. Moreover, most DCDHF molecules have amphiphilic structures, with a polar dicyanomethylenedihydrofuran headgroup and nonpolar hydrocarbon tails on the amine or furan ring, and can be used as fluorescent lipid analogues for live cell imaging. Here we demonstrate that individual molecules of several different DCDHF lipid analogues can be observed diffusing in the plasma membrane of Chinese hamster ovary cells. The photophysical and diffusive behaviors of the DCDHF lipid analogues in membranes are described and are found to be competitive with the well-known lipid probe N-(6-tetramethylrhodaminethiocarbamoyl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine.
Nishimura
Stefanie Y
SY
Department of Chemistry and Biophysics Program, Stanford University, Stanford, CA 94305-5080, USA.
Lord
Samuel J
SJ
Klein
Lawrence O
LO
Willets
Katherine A
KA
He
Meng
M
Lu
Zhikuan
Z
Twieg
Robert J
RJ
Moerner
W E
WE
D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000737:chemistry*; D002784:Cholesterol / Q000737:chemistry; D006224:Cricetinae; D003412:Cricetulus; D003627:Data Interpretation, Statistical; D004058:Diffusion; D005456:Fluorescent Dyes / Q000737:chemistry*; D005663:Furans / Q000737:chemistry*; D007202:Indicators and Reagents; D008563:Membrane Lipids / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D009570:Nitriles / Q000737:chemistry*; D010777:Photochemistry; D017785:Photons; D013109:Sphingomyelins / Q000737:chemistry
D005456:Fluorescent Dyes; D005663:Furans; D007202:Indicators and Reagents; D008563:Membrane Lipids; D009570:Nitriles; D013109:Sphingomyelins; C521333:dicyanomethylenedihydrofuran; D002784:Cholesterol