Enhanced accuracy of single-molecule diffusion measurements with a photocleavable energy-transfer dyad.
A photocleavable energy-transfer dyad was synthesized, characterized, and applied to single-molecule fluorescence microscopy. After photocleavage, a combination of independent two-color single-molecule tracking and analysis of single-molecule energy-transfer efficiencies allows the determination of the temporal evolution of the relative distances between both fragments from the nm to the μm scale. This gives access to a broad range of diffusion coefficients.
Fachbereich Chemie, Universität Konstanz, Universitätsstrasse 10, 78457 Konstanz (Germany).
Maren
M
Dill
Moritz C
MC
Baier
Stefan
S
Mecking
Dominik
D
Wöll
D000873:Anthracenes / Q000138:chemical synthesis* / Q000737:chemistry; D004058:Diffusion; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D007094:Imides / Q000737:chemistry; D008856:Microscopy, Fluorescence; D010569:Perylene / Q000138:chemical synthesis* / Q000737:chemistry; D010782:Photolysis; D014466:Ultraviolet Rays
D000873:Anthracenes; D005456:Fluorescent Dyes; D007094:Imides; C528989:terrylene; D010569:Perylene
Förster resonance energy transfer; diffusion; fluorescence microscopy; photolabile groups; single-molecule spectroscopy