Fluorescence Recovery after Merging a Droplet to Measure the Two-dimensional Diffusion of a Phospholipid Monolayer

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We introduce a new method to measure the lateral diffusivity of a surfactant monolayer at the fluid-fluid interface, called fluorescence recovery after merging (FRAM). FRAM adopts the same principles as the fluorescence recovery after photobleaching (FRAP) technique, especially for measuring fluorescence recovery after bleaching a specific area, but FRAM uses a drop coalescence instead of photobleaching dye molecules to induce a chemical potential gradient of dye molecules. Our technique has several advantages over FRAP: it only requires a fluorescence microscope rather than a confocal microscope equipped with high power lasers; it is essentially free from the selection of fluorescence dyes; and it has far more freedom to define the measured diffusion area. Furthermore, FRAM potentially provides a route for studying the mixing or inter-diffusion of two different surfactants, when the monolayers at a surface of droplet and at a flat air/water interface are prepared with different species, independently.
Publisher
JOURNAL OF VISUALIZED EXPERIMENTS
Issue Date
2015-10
Language
English
Article Type
Article
Keywords

TRANSLATIONAL DIFFUSION; LATERAL DIFFUSION; MEMBRANE; MICRORHEOLOGY; CHOLESTEROL; SURFACTANT; FLUIDITY; LIPIDS

Citation

JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, no.104

ISSN
1940-087X
DOI
10.3791/53376
URI
http://hdl.handle.net/10203/207860
Appears in Collection
BiS-Journal Papers(저널논문)CBE-Journal Papers(저널논문)
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