Phase Imaging of Phosphatidylcholine Bilayer Membranes by Prodan Fluorescence

Prodan (6-propiponyl-2-(N,N-dimethylamino)naphthalene) is well known as a polarity-sensitive fluorescent probe and has a high capability of detecting structural changes occurring within phospholipid bilayer membranes. In this study, we carried out the fluorescence spectroscopic observation of bilayer phase behavior for a series of symmetric saturated diacylphosphatidylcholines (CnPCs) with different acyl-chain length n (n = 12-15 and 19-22) using Prodan as a membrane probe to confirm the availability of Prodan along with the previous results for the CnPC bilayer membranes (n = 16-18). The results were discussed by constructing spectral three-dimensional (3D) imaging plots for visualizing the change in bilayer phase states with temperature or pressure to verify the functionality of this 3D imaging plot. It was found that the Prodan fluorescence technique is applicable to the detection of the changes in the bilayer phase states of all CnPCs with a few exceptions. One of the most crucial exceptions was that Prodan cannot be used for the detection of the bilayer-gel state of the C21PC bilayer membrane. It was also found that it is only to the CnPC bilayer membranes with n = 15-18 that the 3D imaging plot is adequately and accurately applicable as a useful graphical tool for visually detecting the bilayer phase states. This is a disadvantageous feature of this technique brought about by the high sensitivity of Prodan as a membrane probe. Further detailed studies on the molecular behavior of Prodan will enable us to find a more useful way of utilizing this membrane probe.