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Updated: Feb 25, 2026
Fluorescence Recovery after Merging a Droplet to Measure the Two-dimensional Diffusion of a Phospholipid Monolayer
Published on: October 15, 2015
Communication: Development of standing evanescent-wave fluorescence correlation spectroscopy and its application to
Takuhiro Otosu1, Shoichi Yamaguchi1
1Department of Applied Chemistry, Graduate School of Science and Engineering, Saitama University, 255 Shimo-Okubo, Sakura, Saitama 338-8570, Japan.
Abstract:
We present standing evanescent-wave fluorescence correlation spectroscopy (SEW-FCS). This technique utilizes the interference of two evanescent waves which generates a standing evanescent-wave. Fringe-pattern illumination created by a standing evanescent-wave enables us to measure the diffusion coefficients of molecules with a super-resolution corresponding to one fringe width. Because the fringe width can be reliably estimated by a simple procedure, utilization of fringes is beneficial to quantitatively analyze the slow diffusion of molecules in a supported lipid bilayer (SLB), a model biomembrane formed on a solid substrate, with the timescale relevant for reliable FCS analysis. Furthermore, comparison of the data between SEW-FCS and conventional total-internal reflection FCS, which can also be performed by the SEW-FCS instrument, effectively eliminates the artifact due to afterpulsing of the photodiode detector. The versatility of SEW-FCS is demonstrated by its application to various SLBs.
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