Effects of using a metal layer in total internal reflection fluorescence microscopy

Abstract

This proceeding examines the characteristics of imaging through a metal-coated glass cover slip using total internal reflection fluorescence microscopy (TIRFM). Through back and front focal plane imaging of sub-diffraction-limited fluorescent beads, the experimental characteristics of the emission are compared with theoretical simulations. Furthermore, at the angle at which surface plasmon resonance occurs, we show that the anisotropic emission of the fluorescent beads collected through the metal layer results in a irregular point spread function that has a donut-like structure with multiple concentric rings. © 2007 Springer-Verlag.