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Refining membrane penetration by a combination of steady-state and time-resolved depth-dependent fluorescence quenching
- Kyrychenko, Alexander, Ladokhin, Alexey S.
- Analytical biochemistry 2014 v.446 pp. 19-21
- fluorescence, fluorescent dyes, lipid bilayers, refining
- Accurate determination of the depth of membrane penetration of a fluorescent probe, attached to a lipid, protein, or other macromolecule of interest, using depth-dependent quenching methodology is complicated by thermal motion in the lipid bilayer. Here, we suggest that a combination of steady-state and time-resolved measurements can be used to generate a static quenching profile that reduces the contribution from transverse diffusion occurring during the excited-state lifetime. This procedure results in narrower quenching profiles, compared with those obtained by traditional measurements, and thus improves precision in determination of the underlying depth distribution of the probe.