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

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