Pulsed-interleaved excitation FRET measurements on single duplex DNA molecules inside C-shaped nanoapertures.
Single-molecule fluorescence resonant energy transfer (FRET) is a widely accepted method for determining the spatial separation between molecules. In combination with pulsed interleaved excitation (PIE), additional information about the stoichiometry of molecular interactions is obtained. PIE-FRET, however, as implemented with standard confocal optics, requires the dilution of the sample to biologically low concentrations. Here, we show that PIE-FRET measurements inside nanometer-sized apertures yield meaningful biochemical data at 1000 x higher concentrations.
Fore
Samantha
S
NSF Center for Biophotonics Science and Technology, University of California at Davis, 2700 Stockton Boulevard, Suite 1400, Sacramento, California 95817, USA. srfore@ucdavis.edu
Yuen
Yin
Y
Hesselink
Lambertus
L
Huser
Thomas
T
D004247:DNA / Q000737:chemistry* / Q000648:ultrastructure*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D046911:Macromolecular Substances / Q000737:chemistry; D008422:Materials Testing; D008832:Microchemistry / Q000379:methods*; D008968:Molecular Conformation; D049329:Nanostructures / Q000737:chemistry* / Q000648:ultrastructure*; D036103:Nanotechnology / Q000379:methods*; D010316:Particle Size; D013499:Surface Properties
D046911:Macromolecular Substances; D004247:DNA