Surface plasmon-mediated fluorescence microscopy: A powerful imaging technique for membrane imaging

An alternative technique to total internal reflection fluorescence microscopy (TIRFM) is presented, which takes advantage of surface plasmon (SP) properties of a metallic thin-film. SP mediated-excitation and emission together with near-field coupling to fluorophores provide many advantages: enhanced fluorescence signal, increased confinement, and reduction of the photobleaching as well as of the background noise. The two techniques are compared and, contrary to TIRFM, the penetration depth in surface plasmon-mediated fluorescence microscopy (SPMFM) is homogeneous and well defined over the entire image field. SPMFM coupled fluorescence lifetime imaging allows precise membrane/surface reported.