We propose a novel fluorescence emission difference microscopy (FED) system based on focusing cy-

lindrical vector beams. In conventional FED, a Gaussian beam and a 0–2 π vortex phase plate are used to

generate solid and hollow spots. We focus radially polarized and azimuthally polarized cylindrical vector

beams to obtain an expanded solid spot and a shrunken hollow spot, taking advantage of the optical

properties of cylindrical vector beams to improve the conventional FED performance. Our novel method

enhances FED performance