I tried some FCS experiments on DPPC bilayer(bodipy c12hpc - 10^-4 mol%). But the intensity is just decaying over t ( time trace).Is it that gel phase of DPPC is a reason?
I am preparing these bilayers using langmuir blodgett : transfer pressure 35mN/m and subphase temperature - 15C
When performed similar type of experiment on DMPC bilayer(similar pressure and temperature in LB), It worked very well.I got good time trace and correlation curves.
Recently, I stumbeled about this paper which deals with FCS and DPPC - maybe it helps you?
Article Diffusion and Partitioning of Fluorescent Lipid Probes in Ph...
Though in the Gudmand et al. work the main problem was phase separation of the lipid interface, I believe the problem your are facing at the moment is the gel phase. If you would compare Tm of DMPC (~24oC) to DPPC (~41oC) you would see that the difference is pretty striking. Since in gel phase practically any diffusion is allowed you would probably just bleach fluorescent probe.
You might have a look at those two publication in addition to Gudmand et al. They treat on such challenging measurement with some kind of FCS variation.
Article Accurate Determination of Membrane Dynamics with Line-Scan FCS
Article Role of ceramide in membrane protein organization investigat...
In Gudmand's monolayer experiment, they have measured diffusivity for DPPC only at 5mN/m.
Thank you Leonhard and Grzegorz.
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