Resolution enhancement of confocal fluorescence microscopy via two illumination beams

Confocal fluorescence microscopy is an effective imaging technique, but its resolution is limited by the diffraction-limit. Fluorescence emission difference (FED) method is a useful way to improve the resolution of confocal fluorescence microscopy, but the negative values generated during subtraction process might cause loss of valid information. In this paper, we propose one effective method to enhance the resolution of confocal fluorescence microscopy without generating significant negative values. The proposed method combines digital processing and FED method, obtaining the final images with higher resolution and less information loss. © 2019