Abstract. Planar glass waveguides with a specific number of modes were fabricated by Ag+–Na+ exchange in Schott SG11 glass. The effective refractive indices were determined using m-line spectroscopy in both s- and p-polarization. By using the reversed Wentzel–Kramers– Brillouin approximation, the index profiles were described by a nonlinear diffusion equation. The diffusion coefficients for Ag+ were established, as well as the penetration depth of the evanescent field in an aqueous environment for the different modes. The integrals of E2 fields for the evanescent-guided fields were investigated. These are important when evanescent fields are used for illumination in interface microscopy, an alternative method to total internal reflection fluorescence (TIRF) microscopy. The photoluminescent behavior of the waveguides was investigated as a function of ion exchange time and excitation wavelengths. Comparable images were obtained of fluorescently labeled HEK293 cells using TIRF microscopy and waveguide evanescent field fluorescence microscopy. Imaging was performed using HEK293 cells, delivering similar images and information. © 2010 Society of Photo-Optical Instrumentation Engineers.