Surface Plasmon Enhanced TIRF Imaging

Abstract

Total Internal Reflection Fluorescence Microscopy (TIRFM) is a powerful technique for imaging dynamic membrane events in living cells. However the information gained from TIRFM may be limited by the intensity of the florescence signal and by background noise emanating from the inner part of the cell. Here we describe how Surface Plasmon Mediated Fluorescence Microscopy (SPMFM) can enhance TIRFM by increasing the fluorescence signal 5-6 fold and by reducing background noise dramatically. The imaging configuration is that for TIRFM except that the coverglass is coated on one side with a nanometre film of silver. For SPMFM (as in TIRFM), a high numerical aperture (N.A.) objective is essential for achieving required illumination angles.